a putative protein o-fucosyltransferase facilitates pollen ...feb 21, 2018 · 20 one sentence...

1

Short title: AtOFT1 facilitates pollen tube penetration 1

Title: A putative protein O-fucosyltransferase facilitates pollen tube penetration 2

through the stigma–style interface 3

4

Devin K. Smith1, Danielle M. Jones

1, Jonathan B. R. Lau

1, Edward R. Cruz

1, Elizabeth 5

Brown1, Jeffrey F. Harper

1, and Ian S. Wallace

1,2* 6

1Department of Biochemistry and Molecular Biology and

2Department of Chemistry, 7

University of Nevada, Reno, Reno, NV 89557 8

9

*Corresponding author: 10

Ian S. Wallace 11

Department of Biochemistry and Molecular Biology 12

University of Nevada, Reno 13

1664 N. Virginia St. MS0330 14

Reno, NV 89557 15

Email: [email protected] 16

Phone: 775-682-7311 17

Fax: 775-784-4090 18

19

One sentence summary: A putative protein O-fucosyltransferase in Arabidopsis 20

facilitates pollen tube penetration through stigmatic tissue, suggesting that protein O-21

fucosylation events may play a role in plant reproduction. 22

23

Author contributions: Jeffrey F. Harper and Ian S. Wallace conceived the original 24

research and supervised the experiments. Devin K. Smith, Danielle M. Jones, and 25

Elizabeth Brown performed the experiments and were assisted by Jonathon Lau. Devin 26

K. Smith, Danielle M. Jones, Jeffrey F. Harper, and Ian S. Wallace wrote the manuscript. 27

28

29

30

31

32

33

34

35

36

Plant Physiology Preview. Published on February 21, 2018, as DOI:10.1104/pp.17.01577

Copyright 2018 by the American Society of Plant Biologists

https://plantphysiol.orgDownloaded on February 4, 2021. - Published by Copyright (c) 2020 American Society of Plant Biologists. All rights reserved.

mailto:[email protected]

https://plantphysiol.org

2

37

38

Abstract: 39

During pollen–pistil interactions in angiosperms, the male gametophyte (pollen) 40

germinates to produce a pollen tube. To fertilize ovules located within the female pistil, 41

the pollen tube must physically penetrate specialized tissues. Whereas the process of 42

pollen tube penetration through the pistil has been anatomically well-described, the 43

genetic regulation remains poorly understood. In this study, we identify a novel 44

Arabidopsis thaliana gene, O-FUCOSYL TRANSFERASE 1 (AtOFT1), which plays a key 45

role in pollen tube penetration through the stigma–style interface. Semi-in vivo growth 46

assays demonstrate that oft1 mutant pollen tubes have a reduced ability to penetrate the 47

stigma–style interface, leading to a nearly 2000-fold decrease in oft1 pollen transmission 48

efficiency and a 5–10 fold decreased seed set. We also demonstrate that AtOFT1 is 49

localized to the Golgi apparatus, indicating its potential role in cellular glycosylation 50

events. Finally, we demonstrate that AtOFT1 and other similar Arabidopsis genes 51

represent a novel clade of sequences related to metazoan protein O-fucosyltransferases, 52

and that mutation of residues that are important for O-fucosyltransferase activity 53

compromise AtOFT1 function in vivo. The results of this study elucidate a physiological 54

function for AtOFT1 in pollen tube penetration through the stigma–style interface and 55

highlight the potential importance of protein O-glycosylation events in pollen-pistil 56

interactions. 57

58



3

59

Introduction 60

Double fertilization in angiosperms precisely orchestrates the controlled 61

interaction between two non-motile sperm cells, the egg cell, and the central cell, which 62

results in the generation of a new embryo and surrounding endosperm tissues. Initial 63

pollen grain interaction with stigmatic papillae leads to pollen grain hydration and 64

subsequent formation of a specialized structure called the pollen tube, which serves as a 65

vehicle to transport two sperm nuclei to the distant ovule (Palanivelu and Johnson, 2010; 66

Beale and Johnson, 2013). The pollen tube must penetrate through the stigma–style 67

interface (Elleman et al., 1992; Jiang et al., 2005), rapidly elongate within the 68

transmitting tract (Sessions and Zambryski, 1995), and respond to positional guidance 69

cues that lead the pollen tube to the ovule (Palanivelu et al., 2003; Okuda et al., 2009; 70

Takeuchi and Higashiyama, 2012; Takeuchi and Higashiyama, 2016; Mizukami et al., 71

2016). At the ovule, the pollen tube must navigate through the micropylar opening, 72

penetrate through the synergid cell (Sandaklie-Nikolova et al., 2007), and finally rupture 73

to release the sperm cells in which gamete fusion follows (Sandaklie-Nikolova et al., 74

2007; Leydon et al., 2015). 75

The pollen tube must circumvent numerous physical barriers to fertilize the 76

female gamete, and the stigma–style interface represents the first barrier encountered 77

during this process. Ultrastructural microscopy studies of Arabidopsis pollen-pistil 78

interactions indicate that the elongating pollen tube penetrates through the papillar cell 79

cuticle and cell wall, and subsequently grows toward the base of this cell along the 80

surface of the plasma membrane (Elleman et al., 1992; Jiang et al., 2005). After exiting 81

the papillar cell, the pollen tube continues to grow through the middle lamella layer 82



4

between cells in the stylar tissue and eventually enters the transmitting tract (Sessions and 83

Zambryski, 1995; Jiang et al., 2005). While these steps are essential for successful 84

fertilization, relatively few genes involved in pollen tube penetration have been 85

identified, despite the fact that this process markedly alters the pollen tube transcriptome 86

(Qin et al., 2009). 87

Most plant cells adhere to one another throughout the lifetime of the plant due to 88

interactions between adjacent cell walls (Bouton et al., 2002; Durand et al., 2009; 89

Neumetzler et al., 2012; Verger et al., 2016), but the growing pollen tube must modulate 90

cellular adherence as it interacts with multiple cell types during pollen tube penetration. 91

Therefore, pollen-pistil interactions represent a unique opportunity to investigate the 92

understudied process of cell adhesion in plant systems (Chae and Lord, 2011). Very few 93

factors regulating plant cell adhesion have been described. The disruption or structural 94

alteration of pectic cell wall polysaccharides, which form the middle lamella between 95

adherent cells, causes clear defects in cellular adhesion (Bouton et al., 2002; Durand et 96

al., 2009), suggesting that pectins play an important role in plant cell adhesion. Recently, 97

genetic analyses in Arabidopsis implicated FRIABLE1 (FRB1; Neumetzler et al., 2012) 98

and ESMERELDA1 (ESMD1; Verger et al., 2016) as novel regulators of plant cell 99

adhesion, based on the observations that frb1 mutants exhibit cell adhesion-related 100

phenotypes, such as organ dissociation and separation. Furthermore, esmd1 loss-of-101

function mutants suppressed these phenotypes and the phenotypes of other known cell 102

adhesion mutants (Verger et al., 2016), suggesting that FRB1 and ESMD1 play opposing 103

roles in the regulation of plant cell adhesion. Interestingly, both genes encode predicted 104



5

protein O-fucosyltransferases, which are known regulators of cell adhesion and cell-cell 105

communication in animal systems. 106

In metazoan systems, cell adhesion can directly regulate cell-cell communication 107

and cell fate through the Notch signaling cascade (Kovall et al., 2017). Notch family 108

receptors contain a large extracellular domain composed of multiple Epidermal Growth 109

Factor (EGF) repeat domains (Rebay et al., 1991), as well as an intracellular domain that 110

regulates transcription in response to the extracellular domain binding cognate protein 111

ligands, such as Serrate, Delta, and Jagged. The Notch extracellular domain EGF repeats 112

are heavily glycosylated with fucose-containing glycans attached to Serine/Threonine 113

residues (Shi and Stanley, 2003; Okajima et al., 2003; Sasamura et al., 2003; Rampal et 114

al., 2005). EGF fucosylation is catalyzed by protein O-fucosyltransferase1 (POFT1), 115

which uses GDP-fucose as a sugar nucleotide donor to attach fucose to protein targets in 116

the consensus amino acid sequence CXXXX(S/T)C (Wang et al., 2001). These post-117

translational glycosylation events potentiate the interaction of Notch with its cognate 118

ligands (Okajima et al., 2003; Stahl et al., 2008). Recently, the X-ray structure of a 119

glycosylated Notch subdomain bound to Jagged was determined, and this structure 120

demonstrated that O-fucosylated residues participate in “catch” bonds that communicate 121

mechanical information between cells (Luca et al., 2017). Protein O-fucosylation also 122

regulates a number of other metazoan proteins that contain EGF or Thrompospondin 123

Repeat (TSR) domains (Harris and Spellman, 1993; Leonhard-Melief and Haltiwanger, 124

2010). Whereas this modification plays a well-established role in the regulation of 125

metazoan cell adhesion, it has been relatively understudied in plant systems. 126



6

In this study, we identify and characterize a novel Arabidopsis gene, At3g05320, 127

which encodes an O-fucosyltransferase family protein that plays a critical role in pollen 128

tube penetration through the pistil. Phylogenetic analysis showed that this gene was more 129

closely related to metazoan POFT1s than previously described plant POFTs. We 130

therefore named this gene Arabidopsis O-FUCOSYLTRANSFERASE 1 (AtOFT1). 131

Genetic analyses revealed that oft1 mutants exhibit a marked reduction in overall fertility 132

despite the fact that oft1 pollen tubes grow normally under in vitro conditions. Further 133

analysis revealed that oft1 mutant pollen tubes are compromised in their ability to 134

penetrate the stigma–style interface and elongate through the transmitting tract, 135

suggesting that AtOFT1 may play a role in modulating cell adhesion during these events. 136

137

Results 138

Phylogenetic relationship between plant and metazoan protein O-fucosyltransferases 139

To examine the phylogenetic relationship between plant and metazoan POFT1s, 140

the full length amino acid sequences of M. musculus, D. melanogaster, D. rerio, C. 141

elegans, and H. sapiens POFT1 were used as BLAST queries against the Arabidopsis 142

genome. In each case, a previously uncharacterized Arabidopsis gene (At3g05320) 143

exhibited weak (18–21%) amino acid sequence identity to these metazoan POFT1s. In 144

contrast, the queried metazoan POFT1s did not identify previously characterized putative 145

Arabidopsis POFTs, such as FRB1 (Neumetzler et al., 2012) and ESMD1 (Verger et al., 146

2016). To further understand the phylogenetic relationships between At3g05320, other 147

putative Arabidopsis POFTs, and established metazoan POFT1s, a phylogenetic tree 148

containing these protein sequences was constructed (Figure 1). The Arabidopsis genome 149

contains 38 proteins that are annotated as putative POFTs, in agreement with previous 150



7

analyses (Neumetzler et al., 2012). At3g05320 was most phylogenetically similar to a 151

subgroup of POFTs that included three other unidentified Arabidopsis genes (At1g53770, 152

At1g17270, and At5g50420), and the metazoan POFT1 family. Interestingly, these 153

protein sequences preferentially clustered with metazoan POFT1s and were only distantly 154

related to previously identified Arabidopsis putative POFTs, suggesting that At3g05320 155

and its homologues are more similar to metazoan POFT1s in amino acid sequence and 156

potentially in enzymatic function. In light of this phylogenetic similarity, the At3g05320 157

gene was named Arabidopsis thaliana O-FUCOSYLTRANSFERASE 1 (AtOFT1). 158

159

AtOFT1 T-DNA mutants exhibit altered silique morphology and reduced seed set 160

Based on the observation that the AtOFT1 sequence is closely related to metazoan 161

POFT1s, and that the function of AtOFT1 had not been previously studied, a genetic 162

approach was pursued to examine the physiological role of this gene. Three independent 163

AtOFT1 T-DNA insertions were identified (Supplemental Figure S1A) in the Arabidopsis 164

T-DNA insertional mutant database (Alonso et al., 2003), and homozygous T-DNA 165

insertions in these mutants were verified by PCR genotyping (Supplemental Figure S1B). 166

These insertional mutant alleles were renamed oft1-1 (SALK_072442), oft1-2 167

(SALK_151675), and oft1-3 (WiscDsLox489-492M4) (Supplemental Figure S1A). RT-168

PCR analysis confirmed that the AtOFT1 transcript was not expressed in these mutants, 169

suggesting that each of these T-DNA lines contain null alleles (Supplemental Figure 170

S1C). 171

oft1 mutant lines were phenotypically evaluated at various stages of Arabidopsis 172

development. At the reproductive stage, oft1 mutants flowered normally, but developed 173



8

65% shorter siliques compared to those of wild-type Col-0 control plants (Figure 2A–174

2C). Seed sets of the oft1 mutants were examined by clearing fully developed siliques 175

and quantifying the number of seed produced. In contrast to wild-type Col-0 control 176

plants, which produced 45–50 seeds per silique, oft1 mutants produced 5–10 fold fewer 177

seeds (Figure 2D and 2E), indicating that these mutants are reproductively compromised. 178

179

The male oft1 gamete is responsible for reduced reproductive success 180

The observed reproductive defects in oft1 mutants could result from impaired 181

male or female gametophytes. Therefore, the transmission of oft1 mutant alleles in self-182

fertilization and outcross events was examined. The oft1-1 and oft1-3 alleles respectively 183

contained functional kanamycin and BASTA resistance markers associated with their T-184

DNA insertions. Transmission of the oft1-2 allele was assessed by PCR genotyping. 185

Selfing oft1+/-

mutants would be expected to produce 75% progeny containing the oft1 T-186

DNA insertion, but oft1 mutant alleles were transmitted to only 51% (oft1-1), 47% (oft1-187

2), and 52% (oft1-3) of progeny (Table I). The frequency of homozygotes recovered in 188

these selfing events was also estimated by propagating a population of the progeny and 189

visually phenotyping the plants at the reproductive stage. Homozygous mutants were 190

recovered at much lower frequencies than the expected 33% progeny (oft1-1 [3.7% 191

homozygotes, n = 108], oft1-2 [0.9% homozygotes, n = 108], oft1-3 [1.4% homozygotes, 192

n = 72]), indicating that while homozygous mutants can be obtained from self-193

fertilization events, they are much less common than expected by Mendelian inheritance. 194

To further investigate whether this observed segregation distortion was due to 195

defects in the male or female gametophyte, reciprocal crosses were performed using 196



9

oft1+/-

parents, and the transmission of mutant alleles was evaluated in the F1 progeny 197

(Table I). The resulting F1 progeny would be expected to inherit the oft1 allele at a 198

frequency of 50%, and when oft1+/-

pistils accepted Col-0 pollen, 52% (oft1-1) and 51% 199

(oft1-3) of the resulting progeny exhibited herbicide resistance (Table I). In contrast, 200

when oft1+/-

pollen was used to fertilize Col-0 pistils, only 1 transmission event was 201

detected out of 1872 oft1-3 F1 progeny, and no transmission events were observed for 202

oft1-1 F1 progeny, indicating a nearly 2000-fold reduction in oft1 pollen transmission 203

efficiency (Table I). These results suggest that oft1 mutants exhibit segregation distortion 204

specifically attributed to a defective male gamete. 205

To confirm that the near-sterile phenotype of oft1 alleles was due to a loss-of-206

function mutation, a complementation construct was generated consisting of the AtOFT1 207

genomic DNA sequence fused to the N-terminus of GFP under the control of the pollen 208

tube-specific ARABINOGALACTAN PROTEIN 11 (AGP11) promoter (Levitin et al., 209

2008). T1 oft1-3-/-

; 11p::OFT1-GFP+/-

and oft1-3+/-

; 11p::OFT1-GFP+/-

seedlings were 210

selected on hygromcyin as described in Materials and Methods and propagated to 211

flowering stage. This construct, which is referred to as 11p::OFT1-GFP, was introduced 212

into the oft1-3-/-

and oft1-3+/-

genetic backgrounds, and this construct fully or partially 213

complemented the seed set phenotype of oft1-3-/-

mutants in multiple independent 214

transgenic lines (Supplemental Figure S2). These plants were utilized in a variety of 215

outcrossing experiments described below, and the resulting F1 progeny of these crosses 216

were assayed for 11p::OFT1-GFP transmission via hygromycin resistance. 217

Reciprocal crosses were performed with three independent lines harboring a 218

11p::OFT1-GFP+/-

transgene in the oft1-3-/-

background (Table I). In this assay, all of 219



10

the pollen were mutant, and the transmission test evaluated how well the hemizygous 220

pollen harboring the transgene competed with pollen that do not contain the transgene. 221

While an expected 50% transmission was observed through the female gametophyte, a 222

near 100% transmission was observed through the pollen (Table I). These results 223

indicate that oft1-3- pollen tubes containing the 11p::OFT1-GFP transgene significantly 224

outcompete oft1-3- pollen tubes in a direct competition assay. 225

To measure the simultaneous transmission of wild-type, mutant, and 226

complemented AtOFT1 pollen tubes, the transmission of oft1-3- and 11p::OFT1-GFP 227

alleles in outcrosses between Col-0 and oft1-3+/-

; 11p::OFT1-GFP+/-

were evaluated. For 228

these experiments, the BASTA resistance cassette associated with the oft1-3 T-DNA 229

allele was utilized to evaluate transmission. When oft1-3+/-

; 11p::OFT1-GFP+/-

females 230

were used with Col-0 pollen, 50% of the resulting F1 progeny inherited the oft1-3 mutant 231

allele, consistent with the expected value of 50% (Table I). However, when oft1-3+/-

; 232

11p::OFT1-GFP+/-

pollen was used to pollinate Col-0 pistils, only 31% of the F1 233

progeny inherited the oft1-3 T-DNA allele, which was significantly less than the expected 234

50% inheritance (Table I). This result was consistent with the hypothesis that only oft1-235

3-; 11p::OFT1-GFP

+ pollen are able to successfully compete with wild-type pollen 236

(expected value 33%; χ2 = 2.79; P =0.09; Table I), suggesting that oft1-3

mutant pollen 237

tubes could only successfully fertilize ovules if they also contained a functional 238

complement copy of the 11p::OFT1-GFP transgene. Overall, these results provide 239

compelling evidence to support the hypothesis that AtOFT1 plays a significant role in 240

pollen tube physiology and that the male gametophyte is solely responsible for the 241

reduction in oft1 fertility. 242



11

243

AtOFT1 is expressed in pollen tubes 244

To investigate whether AtOFT1 was expressed in developing pollen or other 245

reproductive tissues, public microarray data was examined using the Pollen RCN Heat 246

Tree expression viewer (www.arabidopsis-heat-tree.org). This analysis revealed that 247

AtOFT1 expression was not observed in various female tissues (Supplemental Figure 248

S3A). However, AtOFT1 expression was observed in dry pollen, and transcript 249

abundance increased after pollen tube germination. Interestingly, the highest AtOFT1 250

transcript abundance was observed in 4 h semi-in vivo germinated pollen tubes that had 251

penetrated through dissected stigmatic tissue (Qin et al., 2009), suggesting that AtOFT1 252

expression increases as the pollen tube interacts with the female tissues. This analysis 253

also revealed that At1G53770 and At5g50420, two other genes that are closely 254

phylogenetically related to AtOFT1, exhibited very different expression profiles in 255

reproductive tissues. At1g53770 was strongly expressed in sperm cells, with lower but 256

significant expression in ovule tissue. At5g50420 was expressed throughout the pistil 257

tissue but was not expressed in the male gametophyte tissues after pollen maturity, 258

suggesting that AtOFT1-like OFTs may play distinct roles in Arabidopsis reproductive 259

tissues due to their non-overlapping expression patterns. To additionally confirm 260

AtOFT1 pollen tube expression, 1000 base pairs of the putative AtOFT1 promoter was 261

fused to a GFP reporter (Supplemental Figure S3B), and this construct was transformed 262

into wild-type Col-0 plants. Subsequent fluorescence imaging of pollen derived from T1 263

transformants revealed substantial GFP signal in in vitro germinated pollen tubes 264



12

(Supplemental Figure S3C–F), confirming that AtOFT1 is expressed in growing pollen 265

tubes. 266

267

Subcellular localization of AtOFT1 268

Metazoan POFTs are localized to the endoplasmic reticulum where they modify 269

target substrates before being further glycosylated in the Golgi apparatus (Luo and 270

Haltiwanger, 2005). To further understand the function of AtOFT1, the previously 271

described 11p::OFT1-GFP transgenic lines were used to investigate the subcellular 272

localization of this protein. Pollen tubes were germinated under in vitro conditions and 273

examined by confocal microscopy 1.5 h after germination. AtOFT1-associated GFP 274

signal was localized to punctate motile intracellular organelles (Figure 3C, Supplemental 275

Movie 1). 276

To further investigate this subcellular localization pattern, the predicted 277

subcellular localization of AtOFT1 was examined using the SUBcellular localization 278

database of Arabidopsis proteins (SUBA; Tanz et al., 2013). AtOFT1 was predicted to 279

be mitochondrially-localized by SUBA, so 11p::OFT1-GFP expressing pollen tubes were 280

germinated in vitro and stained with 500 nM MitoTracker Orange. As shown in Figure 281

3A, AtOFT1-GFP signal did not overlap with MitoTracker Orange-stained mitochondria, 282

and quantitative co-localization analysis using JACoP (Bolte and Cordelieres, 2006) 283

revealed a Pearson’s correlation coefficient of only 0.21 between the AtOFT1-GFP and 284

MitoTracker Orange signals (Figure 3B), suggesting that AtOFT1 is not localized to 285

pollen tube mitochondria as predicted. 286



13

Subcellular localization studies previously determined that FRB1 and ESMD1 287

were localized to the Golgi apparatus in interphase cells (Neumetzler et al., 2012; Verger 288

et al., 2016), suggesting that AtOFT1 may also localize to this organelle. To test this 289

hypothesis, the 11p::OFT1-GFP transgenic construct was introgressed into transgenic 290

plants expressing the established Golgi markers MEMBRIN12 (MEMB12) or GOLGI 291

TRANSPORT1 (Got1p) fused to mCherry (Geldner et al., 2009). Pollen harvested from 292

6-week-old F1 progeny was germinated in vitro and examined by confocal fluorescence 293

microscopy. These experiments revealed significant localization overlap between 294

AtOFT1-GFP and Golgi marker fluorescent signals (Figure 3A). Quantitative co-295

localization analyses revealed that both the MEMB12- and Got1p-mCherry markers 296

respectively co-localized with AtOFT1-GFP signal with a Pearson’s correlation 297

coefficients of 0.79 and 0.72 (Figure 3B), suggesting a high degree of co-localization 298

between AtOFT1-GFP and known Golgi-resident proteins. These results suggest that 299

AtOFT1 localizes to the Golgi apparatus and potentially participates in cellular 300

glycosylation events in this organelle. 301

302

AtOFT1 facilitates pollen tube penetration through the stigma–style interface 303

The reciprocal outcross results suggested that oft1 mutants were specifically 304

compromised in some aspect of pollen tube physiology. Mutations that compromise 305

pollen tube elongation often exhibit reduced fertility, so oft1 mutant pollen was 306

germinated in vitro to compare their relative elongation rates and morphology to wild-307

type Col-0 control pollen. As illustrated in Figure 4A, in vitro germinated pollen tubes 308

from all three oft1 mutant lines appeared phenotypically indistinguishable from wild-type 309



14

Col-0 pollen tubes in terms of overall length and morphology. Pollen tube growth rates 310

from the oft1 mutant lines and wild-type Col-0 control pollen tubes were further 311

evaluated (Figure 4B). This analysis revealed that under in vitro conditions, oft1 mutant 312

pollen tubes grew at similar rates to control pollen tubes. To further compare pollen tube 313

elongation rates, pollen harvested from oft1-3-/-

; 11p::OFT1-GFP+/-

was germinated and 314

visualized using both epifluorescent and brightfield microscopy, facilitating the 315

simultaneous examination of rescued and non-rescued pollen tube elongation rates. 316

Pollen germination rates of complemented and oft1 mutant pollen tubes were 317

indistinguishable (Supplemental Figure S4A). As shown in Supplemental Figure S4B, 318

pollen tube growth rates of oft1-3-; 11p::OFT1-GFP

+ and oft1-3

-; 11p::OFT1-GFP

- were 319

also essentially identical, further indicating that pollen tube elongation is not 320

compromised in the oft1 mutant. Overall, these observations indicate that oft1 mutant 321

pollen tubes germinate and develop normally, suggesting that these factors cannot explain 322

the reduced transmission of oft1 pollen. 323

To further investigate the mechanistic basis of the compromised oft1 reproductive 324

phenotype, pollen tube elongation in the pistil was initially examined. Col-0 or oft1-1-/-

325

mutant pollen was applied to intact, emasculated male sterile 1 (ms1; Wilson et al., 2001) 326

mutant pistils. These pistils were harvested after 24 h and stained with analine blue as 327

described in Materials and Methods. Subsequent fluorescence microscopy imaging 328

revealed that Col-0 pollen tubes had largely traversed the stigma–style interface and 329

extended into the ovary cavity (Figure 5A top). In contrast, oft1-1 pollen tubes were 330

largely retained in the stigma–style interface at this time point (Figure 5A bottom). 331



15

To more closely examine this phenomenon, a semi-in vivo assay was utilized to 332

examine the initial stages of pollen tube growth at the stigma–style interface. 333

Emasculated ms1 mutant pistils were pollinated with Col-0 control or homozygous oft1 334

mutant pollen, dissected from the remainder of the pistil 20 min after pollination and 335

maintained on a media surface as described in Materials and Methods. Pollinated semi- 336

in vivo stigmas were examined over time to determine the rate of pollen tube emergence 337

from the transmitting tract (TT) and the total number of pollen tubes exiting the TT. In 338

four independent trials, the percentage of stigmas that exhibited at least one pollen tube 339

exiting the TT over the course of the assay was quantified. Wild-type Col-0 pollen tubes 340

exited the TT of 50% of the stigmas by 3 h after pollination (HAP), and 100% of stigmas 341

pollinated with Col-0 pollen exhibited pollen tubes exiting the TT by 3.5–4.0 HAP 342

(Figure 5B and 5C). In contrast, oft1 mutant pollen tubes had only exited the TT of 5% 343

of stigmas by 3 HAP, and only 50–75% of oft1 pollinated stigmas exhibited pollen tube 344

exit 4.5 HAP (Figure 5B and 5C). The number of pollen tubes exiting the TT over time 345

was also quantified. Wild-type Col-0 pollen tubes were first observed exiting the TT at 3 346

HAP, and a linear increase in pollen tube number was observed over the course of the 347

experiment to a maximum of approximately 40 pollen tubes at 8 HAP (Figure 5D). In 348

contrast, oft1 mutant pollen tubes exhibited a substantial temporal delay in exiting the 349

TT. Approximately 5 pollen tubes were observed exiting the TT at 4 HAP, and a 350

maximum of 12 pollen tubes were observed 8 HAP (Figure 5D). The growth rate of 351

pollen tubes exiting the stigma–style interface was also quantified (Supplemental Figure 352

S5). While Col-0 control pollen tubes exhibited a rapid growth rate that eventually 353

plateaued at 4 HAP, oft1 mutant pollen tubes exhibited a much slower elongation rate 354



16

that remained linear over the course of the assay. These mutant pollen tubes also 355

achieved a much shorter final length. These observations suggest that oft1 mutant pollen 356

tubes are compromised in their ability to penetrate the stigma–style interface, despite 357

their ability to elongate normally under in vitro conditions. 358

To further test the hypothesis that oft1 mutants are unable to efficiently penetrate 359

the stigma–style interface, a semi-in vivo competitive pollen tube penetration assay was 360

also developed. Pollen was harvested from 6-week-old oft1-3-/-

;11p::OFT1-GFP+/-

T1 361

transformants and used to pollinate emasculated ms1 pistils following the aforementioned 362

SIV setup. Four HAP, the dissected stigmas were observed by fluorescence microscopy. 363

Fluorescent pollen tubes containing the 11p::OFT1-GFP transgene and non-fluorescent 364

oft1-3- mutant pollen tubes would be expected to exit the TT at equal frequencies, but 365

fluorescent pollen tubes were observed to exit the TT at a much higher frequency (85–366

90%) (Figure 6A). The proportion of fluorescent and non-fluorescent pollen tubes that 367

did not penetrate the stigma–style interface and elongated away from the stigmatic 368

papillae was also quantified, and 50% of these pollen tubes contained the 11p::OFT1-369

GFP transgene, indicating that the complementation construct was inserted as a single 370

copy in these transgenic lines. As a control, similar semi-in vivo penetration competition 371

assays were performed with pollen expressing a hemizygous copy of Yellow Fluorescent 372

Protein (YFP) under the control of the pollen tube specific AUTOINHIBITED CALCIUM 373

ATPASE 9 (ACA9) promoter, which we refer to as 9p::YFP (Schiott et al., 2004). The 374

pollen tubes emerging from the TT were quantified, and 45% of emerging pollen tubes 375

exhibited 9p::YFP-associated fluorescence (Figure 6B), suggesting that this control 376

transgene had little influence on pollen tube penetration. Overall, these results suggest 377



17

that oft1-3- pollen tubes containing the 11p::OFT1-GFP

+ construct outcompete oft1-3

- 378

mutant pollen tubes for penetration through the stigma–style interface and exit into the 379

TT. Due to the fact that oft1 mutant pollen tubes are capable of normal elongation under 380

in vitro conditions, these results strongly support the hypothesis that oft1 mutant pollen 381

tubes are compromised in their ability to physically penetrate through the stigma–style 382

interface. 383

384

The stigma–style interface is a critical barrier for oft1 pollen tubes 385

While the stigma–style interface is often considered essential for plant sexual 386

reproduction, fertilization events have been observed in the absence of this interface in 387

tobacco, Lilly, and eucalyptus (van Tuyl et al., 1991; Goldman et al., 1994; Trindade et 388

al., 2001). Our results suggest that AtOFT1 plays a significant role in pollen tube 389

penetration through the stigma–style interface, and to test this hypothesis in more detail, 390

we adapted these “decapitation” fertilization assays for use in Arabidopsis. In this assay, 391

mature ms1 flowers were emasculated, and the stigma–style interface was excised for the 392

remainder of the pistil. Pollen derived from oft1-3+/-

parent plants was applied to the 393

dissected pistil (see experimental design in Supplemental Figure S6). The pollen 394

germinated and successfully fertilized ovules, resulting in the production of seed. While 395

this process was inefficient compared to normal outcrosses in the presence of a stigma, 396

sufficient seed (19 ± 7 seeds per silique) was produced in these assays to examine oft1-3 397

mutant allele transmission in the absence of a stigma. Interestingly, transmission of the 398

oft1-3 mutant allele increased dramatically under these conditions, with 29.1% of the 399

resulting progeny inheriting the oft1-3 T-DNA allele (n = 206; TE = 0.41). While this 400



18

result did not reach the 50% progeny that would be expected from Mendelian inheritance, 401

removal of the stigma and style increased transmission efficiency by 773-fold compared 402

to the < 0.1% transmission, which was observed when oft1-3+/-

pollen was utilized to 403

pollinate intact pistils. We additionally postulated that this assay format represented a 404

limited pollination scenario that could yield different transmission rates compared to full 405

pollination experiments carried out above. To examine this possibility, intact ms1 406

stigmas were pollinated with limiting amounts of oft1-3+/-

pollen, and oft1-3 transmission 407

was assayed in the resulting F1 progeny. Under limiting pollination conditions, 2% of 408

the resulting progeny inherited the oft1-3 mutant allele (n = 86), suggesting that oft1 409

mutant pollen are more competitive under limiting pollination conditions, but still 410

ineffective compared to full pollination conditions or pollination in the absence of an 411

intact stigma–style interface. These results suggest that oft1-3 mutant pollen tubes are 412

capable of fertilizing ovules, but that the stigma–style interface represents a critical 413

barrier which slows their penetration through these female tissues. 414

415

Catalytically important POFT1 amino acids impact AtOFT1 function 416

To further examine the amino acid sequence similarities between previously 417

characterized metazoan POFT1s and AtOFT1, we compared the conservation of residues 418

that participate in POFT catalysis between these protein sequences. The X-ray crystal 419

structure of C. elegans POFT1 (CePOFT1) was previously determined in the presence of 420

GDP-Fucose (GDP-Fuc), and this crystal structure illustrates the network of amino acid 421

residues that participate in CePOFT1 substrate selectivity as well as catalysis (Figure 7A) 422

(Lira-Navarette et al., 2011). The crystal structure of CePOFT1 revealed that the β-423



19

phosphate and fucose moieties of GDP-Fuc are coordinated by a critical arginine residue, 424

CePOFT1R240

. CePOFT1N43

was also shown to position the hydroxyl group of the 425

incoming protein substrate in proximity to the fucosyl moiety of GDP-Fuc, hydrogen 426

bond to the fucose ring, and interact with the α-phosphate of GDP-Fuc. CePOFT1R40

427

interacts with the ribose ring of GDP-Fuc and also hydrogen bonds to the fucose ring. 428

Finally, CePOFT1D244

was implicated as a residue which potentially stabilizes the GDP-429

Fuc bound conformation of the enzyme. Importantly, in vitro assays indicated that all of 430

these residues are critical for POFT activity and GDP-Fuc binding (Lira-Navarette et al., 431

2011). Amino acid sequence alignments between AtOFT1 and metazoan POFT1s 432

revealed that many of these critical residues are conserved in AtOFT1 (Figure 7B). 433

AtOFT1R260

aligned with CePOFT1R240

, the critical arginine residue required for 434

catalysis. Similarly, conserved residues corresponding to CePOFT1R40

and D244

were 435

identified (AtOFT1R51

and D264

). Interestingly, CePOFT1N43

was not conserved in 436

AtOFT1, and this residue position contained a histidine substitution. Based on the 437

hydrogen bonding function of this residue in the CePOFT1 mechanism (Lira-Navarette et 438

al., 2011), we postulated that AtOFT1H54

could serve a similar function. 439

To functionally assess the importance of these conserved catalytic residues (R51, 440

H54, R260, and D264), site-directed mutagenesis was performed to individually alter 441

these residues in the vector pUBQ10::OFT1-GFP (see Materials and Methods for a list of 442

mutant constructs). Expression of each site-directed mutant was verified by RT-PCR and 443

by confocal microscopy examination of in vitro germinated pollen tubes to confirm 444

proper subcellular localization (Supplemental Figure S7). The functional importance of 445

each residue was assessed by evaluating the ability of each mutant construct to 446



20

complement the oft1 seed set phenotype (Figure 7C). When residues R51, H54, R260, 447

and D264 were individually mutated to encode alanine, seed set was not significantly 448

restored compared to the homozygous mutant background, which suggested these 449

residues contribute important structural or catalytic roles in AtOFT1. Similar to 450

CePOFT1, an AtOFT1R260K

mutant was also unable to complement the oft1-/-

seed set 451

phenotype, which indicated this residue was fundamentally important in AtOFT1 452

catalysis due to its stringent requirement for an arginine residue at this position. 453

Interestingly, when AtOFT1H54

was mutated to Asparagine, the aligned and catalytically 454

relevant residue in CePOFT1, full complementation of the mutant seed set phenotype was 455

observed. Overall, these observations demonstrate the functional importance of 456

catalytically conserved residues between CePOFT1 and AtOFT1, which additionally 457

suggests these enzymes may share a conserved biochemical mechanism. 458

459

460

Discussion 461

In this study, we describe the novel role of a previously uncharacterized putative 462

protein O-fucosyltransferase from Arabidopsis (AtOFT1). We demonstrate that AtOFT1 463

loss-of-function mutants displays a near-sterile phenotype that causes as much as a 10-464

fold reduction in seed set. In addition, pollen transmission efficiency is reduced by 465

nearly 2000-fold when measured through pollen outcrosses where mutant pollen compete 466

with wild-type for fertilization of ovules (Table I). Interestingly, in vitro germination of 467

oft1 pollen indicates that mutant pollen tubes have similar overall morphology, pollen 468

germination, and pollen tube elongation rates compared to controls (Figure 4; 469



21

Supplemental Figure S4), suggesting that these reproductive defects are not caused by 470

impaired pollen tube growth or abnormal development. However, oft1 mutant pollen 471

tubes exhibited significant delays when penetrating through the stigma–style interface 472

both in intact pistils and during semi-in vivo pollination experiments (Figures 5 and 6), 473

suggesting that AtOFT1 is required for efficient pollen tube penetration through the 474

stigma and style tissues during the early events underlying double fertilization. 475

Additionally, outcross transmission assays utilizing oft1+/-

pollen to fertilize pistils with 476

the stigma–style interface removed indicated that in the absence of this interface, oft1 477

mutant pollen tubes are over 700-fold more successful at fertilizing ovules, suggesting 478

that the stigma–style interface represents a critical barrier to fertilization for oft1 mutants. 479

Overall, these results provide a strong argument that AtOFT1 plays a critical role in 480

pollen tube penetration through the stigma–style interface. 481

The genetic mechanisms underlying pollen tube penetration through the pistil 482

remain unclear (Elleman et al., 1992; Jiang et al., 2005). In this study, we identify 483

AtOFT1 as a gene encoding a putative protein O-fucosyltransferase that facilitates pollen 484

tube penetration through the stigma and style tissues. We postulate that this behavior 485

could be explained by at least three different physiological mechanisms. First, AtOFT1 486

may be required for the pollen tube to respond to currently unidentified positional 487

guidance response cues within the style and transmitting tract. Although responses to 488

specific guidance cues in these tissues must be further explored, in this scenario, oft1 489

mutant pollen tubes could fail to perceive such a stimulus that potentiates their ability to 490

efficiently fertilize ovules. In line with this hypothesis, similar reduced pollen tube 491

penetration behavior has been observed in Arabidopsis VACUOLAR PROTEIN 492



22

SORTING 41 mutants (vps41; Hao et al., 2016). The authors of this study suggested that 493

vps41 plants are not able to turn over unidentified guidance cue receptors, leading to 494

reduced pollen tube growth in the style, ultimately causing reduced pollen tube 495

penetration rates. Second, pollen tubes could penetrate female tissues by simply 496

degrading cell wall material that connects cells within the style and transmitting tract. In 497

this case, AtOFT1 could enable the secretion of cell wall degrading enzymes by the 498

pollen tube (Jiang et al., 2005) or post-translationally glycosylate these enzymes to 499

promote their hydrolytic activity. Finally, a third scheme could be imagined in which 500

AtOFT1 participates in the synthesis of a pollen tube cell wall component that confers 501

mechanical strength during penetration though the female tissues. Indeed, other 502

Arabidopsis putative protein O-fucosyltransferases, such as FRB1, MSR1, and MSR2, 503

display alterations in cell wall architecture and composition (Neumetzler et al., 2012; 504

Verger et al., 2016; Wang et al., 2013). However, oft1 mutant pollen tubes germinated 505

normally and displayed no obvious morphological abnormalities, such as early pollen 506

tube tip swelling or bursting, suggesting that if AtOFT1 does contribute to the penetrative 507

capacitance of the pollen tube cell wall, it must only become necessary in the presence of 508

the female tissues. 509

Protein O-fucosylation and the enzymes that catalyze this post-translational 510

modification are relatively understudied in plant systems (Neumetzler et al., 2012; Verger 511

et al., 2016). Metazoan protein O-fucosyltransferases glycosylate numerous substrates, 512

and these fucosylation events often critically regulate protein-protein interactions 513

involved in cell adhesion and cell-cell communication. Supporting the hypothesis that 514

POFTs perform similar functions in plant cells, the few POFT-like genes that have been 515



23

characterized in plants do demonstrate cell adhesion-related phenotypes. For example, 516

FRB1 plays an important role in cell adhesion during Arabidopsis growth and 517

development, as these mutants exhibit cell sloughing and dissociation, suggesting that 518

this putative POFT plays a role in vegetative cell adhesion throughout the plant 519

(Neumetzler et al., 2012). Interestingly, loss-of-function mutations in ESMD1, a second 520

putative POFT, were recently described as suppressor mutations of frb1 mutant cell 521

adhesion-related phenotypes as well as other genes known to play a role in plant cell 522

adhesion (Verger et al., 2016). These observations suggest that POFT-like glycosylation 523

events may antagonize one another or serve as a form of cell-cell communication during 524

cellular adherence. 525

The substrates of AtOFT1 and all other plant POFT-like plant genes remain 526

unidentified. Metazoan POFT1s utilize GDP-Fuc to fucosylate specific serine or 527

threonine residues in CXXXX(S/T)C consensus sequences within EGF repeat or TSR 528

domains (Wang et al., 2001). A previous study identified protein sequences in the 529

Arabidopsis genome that contained EGF repeat-like domains and noted that TSR-like 530

domains were absent from the Arabidopsis genome (Verger et al., 2016). Very few EGF-531

like domains were identified in this study, and the majority of these proteins were 532

transmembrane receptors, including Wall Associated Kinases (WAKs), which are known 533

to sense pectic cell wall polysaccharides during development and plant defense (Kohorn, 534

2016). It is currently unclear whether AtOFT1 utilizes GDP-Fuc or whether this enzyme 535

fucosylates similar substrates as its metazoan counterparts. However, of the 38 putative 536

POFTs in the Arabidopsis genome, AtOFT1 is most phylogenetically similar to metazoan 537

POFT1 and additionally shares similar residues of critical importance to enzymatic 538



24

activity. Mutagenesis of these critical residues in AtOFT1 leads to loss-of-function in 539

vivo (Figure 7), suggesting that AtOFT1 may perform a similar enzymatic function to 540

metazoan POFT1s. We additionally note that a biochemically-verified POFT was 541

recently described in Arabidopsis (Zentella et al., 2017). Arabidopsis SPINDLY was 542

demonstrated to directly fucosylate specific S/T residues in the DELLA protein, a critical 543

regulator of Gibberelin signaling. These fucosylation events activated the GA signaling 544

pathway by modulating protein-protein interactions between DELLA and other known 545

transcriptional activators of light and brassinosteroid signaling responses. SPINDLY is 546

not phylogenetically related to metazoan POFT1s, suggesting that either metazoan-like 547

POFTs have acquired novel functions in plant systems or that protein O-fucosylation 548

enzymes have evolved multiple times throughout evolution. 549

550

Conclusions: 551

Overall, the results from this study provide strong evidence that AtOFT1 plays a critical 552

role in pollen tube penetration through the stigma and/or style tissues. Additional genetic 553

and cell biological evidence suggests that a Golgi-localized fucosyltransferase system 554

may be required for pollen tube growth through pistil tissues. While the substrates of 555

AtOFT1 are currently unknown, it is possible that these substrates include cell surface 556

receptors or structural proteins that are secreted into the extracellular matrix, and 557

identifying AtOFT1 substrates will be the subject of future studies. The discovery of a 558

near-sterile oft1 phenotype provides a unique opportunity to use robust genetic and cell 559

biological techniques to investigate how POFTs are utilized in plant systems, from 560



25

pollen–pistil interactions during sexual reproduction to overall plant growth and 561

maintenance. 562

563

Materials and Methods 564

Plant growth and maintenance 565

Arabidopsis seeds were sterilized in seed cleaning solution (3% [v/v] sodium 566

hypochlorite, 0.1 % [w/v] sodium dodecyl sulfate) for 20 min at 25°C. Seeds were 567

washed five times in sterile water and incubated at 4°C for 48 h before plating. Seeds 568

were germinated on MS media (1/2× Murashige and Skoog salts, 10 mM MES-KOH pH 569

5.7, 1% [w/v] sucrose, and 1% [w/v] phytoagar) and grown vertically for 10 days under 570

long day conditions (16 h light/8 h dark) at 24°C. These seedlings were transferred to 571

soil and propagated in a Percival AR-66L2 growth chamber under long day conditions 572

until seed set. Arabidopsis outcrosses were performed as described previously (Myers et 573

al., 2009). For seed set imaging, siliques from 6-week-old adult wild-type Col-0 or 574

homozygous oft1 mutant Arabidopsis plants were harvested and incubated for 48 h at 575

25°C in 70% [v/v] ethanol under fluorescent light. Seeds in the cleared siliques were 576

visualized with a Leica EZ4HD video dissecting microscope at 35X magnification. 577

For plant transformations, Agrobacterium tumefaciens GV3101 cells harboring 578

various plant expression constructs were grown in 10 mL Luria Broth (LB) cultures 579

supplemented with 100 µg/ mL spectinomycin, 50 µg/ mL gentamycin, and 25 µg/ mL 580

rifampicin for 16 h at 30°C. These cultures were used to seed 1 L LB cultures containing 581

spectinomycin, gentamycin, and rifampicin at the indicated concentrations above and 582

grown for 16 h at 30°C. Cells were harvested by centrifugation at 3000 × g for 15 min, 583



26

and the resulting pellets were resuspended in 450 mL of 5% [w/v] sucrose supplemented 584

with 0.05% [v/v] Silwet L-77. Five-week-old flowering Arabidopsis plants were 585

transformed via the floral dip method (Clough and Bent, 1998). T1 transformants were 586

identified on selection media (1/2X MS salts, 1% [w/v] sucrose, 0.8% [w/v] phytoagar, 587

10 mM MES-KOH pH 5.7, 100 µg/ mL cefotaxime, 15 µg/ mL BASTA or 25 µg/ mL 588

hygromycin) after a 10-day incubation at 24° under long-day conditions. Resistant 589

seedlings were selected and transferred to soil. 590

591

Isolation of AtOFT1 knockout line 592

AtOFT1 (At3g05320) was queried against the Salk Institute T-DNA insertional 593

mutant database (Alonso et al., 2003), and three potential T-DNA lines were identified 594

(oft1-1, SALK_072442; oft1-2, SALK_151675; and oft1-3, WiscDsLox489-492M4). 595

These seed populations were propagated on MS media as described above, transplanted 596

to soil, and grown under long day conditions at 24°C. Genomic DNA isolation for PCR 597

genotyping was performed essentially as previously described (Edwards et al., 1991) with 598

slight modifications (Villalobos et al., 2015). Each genomic DNA sample was analyzed 599

by PCR genotyping using locus specific primers as well as the appropriate T-DNA 600

specific left border primer (Supplemental Table S1), and ExTaq polymerase (Takara Bio, 601

Mountain View, CA). Reactions were cycled under the following conditions: 95°C 602

initial denaturation for 5 min, 35 cycles of 95°C (30 sec), 52°C (30 sec), 72°C (1.5 min), 603

final extension at 72°C for 7 min. The resulting PCR products were separated on 1.0% 604

(w/v) agarose gels and documented with a Bio-Rad Gel Doc XR+ Image analysis 605



27

workstation. The oft1-3-/-

line was backcrossed to Col-0 to remove a second T-DNA 606

insertion and its associated BASTA resistance marker. 607

608

Cloning of transgenic constructs 609

An OFT1 promoter fragment containing 1000 bp upstream of the predicted start 610

codon and 12 bp after the start codon was cloned using genomic DNA isolated as 611

described above, promoter-specific primers (pOFT1 -1000 pENTR F and pOFT1 pENTR 612

R; Supplemental Table S1), and Phusion DNA polymerase. PCR reactions were cycled 613

under the following conditions: 98°C initial denaturation for 5 min, followed by 35 614

cycles of 98°C (30 sec), 52°C (30 sec), 72°C (2 min), final elongation for 7 min at 72°C. 615

The resulting DNA fragment was resolved on a 1.0 % (v/v) low-melting point agarose 616

gel, excised, and gel purified with the QiaQuick gel extraction kit (Qiagen Valencia, CA). 617

This extracted DNA fragment was cloned into the pENTR-D-TOPO vector according to 618

the manufacturer’s instructions (ThermoFisher Scientific San Jose, CA). The AtOFT1 619

promoter sequence was transferred into the pIGWB-504 binary vector (Nakagawa et al., 620

2007) using LR Clonase II according to the manufacturer’s instructions. 621

The rescue construct 11p::OFT1-GFP encodes AtOFT1-GFP-6xHis-TEV 622

protease site-Halo tag-6xHis. The AtOFT1 genomic sequence was amplified by PCR 623

using gene-specific primers (OFT1 cloning F and R) and transferred into a modified 624

pGreenII vector system for plant expression (Hellens et al., 2000), with a kanamycin 625

selection marker for bacteria, and a hygromycin marker for plants. The DNA sequence of 626

this construct is provided as a supplemental file (Supplemental Figure S8). The 11p 627

promoter corresponds to the upstream regulatory region for AGP11 (At3g01700) with the 628



28

addition of a 5′ UTR region containing an intron from proton pump AHA3 (At5g57350) 629

(Frietsch et al., 2007). The GFP does not contain a S65T modification for enhanced 630

fluorescence in order to maintain a better tolerance of acidic pHs that are often found in 631

compartments in the secretory pathway. The HaloTag® from Promega provides a tag for 632

purification, along with two 6-His tags. 633

Site-directed mutagenesis plasmids were fashioned using the vector 634

pUBQ10::OFT1-GFP, which encodes AtOFT1 with GFP fused to the C-terminus under 635

the control of the Ubiquitin 10 promoter. The AtOFT1 genomic DNA sequence was 636

amplified using gene-specific specific primers (OFT1-pENTR F and OFT1-pENTR R; 637

Table S1). PCR reactions were cycled under the following conditions: 98°C initial 638

denaturation for 5 min, followed by 30 cycles of 98°C (30 sec), 55°C (30 sec), 72°C (1 639

min), final elongation for 5 min at 72°C. The resulting DNA fragment was gel purified 640

and cloned into pENTR-D-TOPO as described above. The AtOFT1 sequence was 641

transferred into the plant compatible Gateway vector, pUBC-GFP (Grefen et al., 2010) 642

using LR Clonase II according to the manufacturer’s instructions. Individual site-643

directed mutagenesis PCR reactions were subsequently assembled using OFT1/ pENTR-644

D-TOPO as a template and nucleotide-specific mutagenic primers (Supplemental Table 645

S1) to create 6 plasmids containing a single altered amino acid residue. PCR reactions 646

were cycled under the following conditions: 98°C initial denaturation for 5 min, 647

followed by 35 cycles of 98°C (30 sec), 60°C (30 sec), 72°C (4.5 min), final elongation 648

for 15 min at 72°C. Following DpnI digestion for 2 h at 37°C, reactions were directly 649

transformed into chemically competent E. coli. 650



29

All constructs were verified by Sanger DNA sequencing at the Nevada Genomics 651

Center (http://www.ag.unr.edu/genomics). Following sequence validation, all constructs 652

were transformed into A. tumefaciencs GV3101 and then transformed into the indicated 653

Arabidopsis background as described using the standard floral dip method (Clough and 654

Bent, 1998). 655

656

Phylogenetic analysis of putative Arabidopsis OFTs 657

The AtOFT1 amino acid sequence was used as a BLAST query to identify 658

putative POFT homologs in the Arabidopsis genome using the WU-BLAST function on 659

The Arabidopsis Information Resource (TAIR) website (www.arabidopsis.org). The 660

amino acid sequence of Mus Musculus (NP_536711.3), Drosophila melanogaster 661

(AAF58290.1), Danio rerio (NP_991283.3), Homo sapiens (NP_056167.1) and 662

Caenorhabditis elegans (ABA29469.1) were obtained from the National Center for 663

Biotechnology Information (NCBI) protein sequence database. Protein sequences were 664

aligned using the multiple alignment mode in ClustalX2 (www.clustal.org), and 665

incomplete sequences were removed. This alignment was used to create a neighbor-666

joining phylogenetic tree consisting of 1000 independent bootstrap trials in MEGA7 667

(http://www.megasoftware.net/). The resulting phylogenetic trees were viewed and 668

analyzed in MEGA7. 669

The molecular model of C. elegans POFT1 was generated using the Visual 670

Molecular Dynamics software (VMD) version 1.9.3. The crystal structure of this protein 671

was generated in complex with GDP-Fucose (GDP-Fuc) by Lira- Navarette et al., 2011 672

and accessed through the Protein Data Bank (PDB ID 3ZY5). 673


http://www.ag.unr.edu/genomics)


30

674

In vitro pollen tube growth assays 675

Pollen was harvested from the opened flowers of 6-week-old plants through 676

gentle blotting action of the floral opening across a 76 × 25mm glass slide containing 677

750L solid Pollen Germination Medium (PGM) (5 mM CaCl2, 0.01% [w/v] boric acid, 678

5 mM KCl, 10% [w/v] sucrose, 1 mM MgSO4, 1.5% [w/v] low melting point agarose, pH 679

7.5) (Boavida and McCormick, 2007) pad cooled to 25ºC. Pollen was germinated in the 680

dark at 25°C in a humidified chamber for the indicated time period. Before imaging, 681

liquid PGM (lacking low melting point agarose) was applied to the surface of the agar 682

media and a coverslip was added. Pollen tube growth and morphology was visualized 683

with a Keyence BZ-X700 microscope under brightfield illumination using a 10X 0.45 684

NA air objective. Pollen tube lengths were quantified using ImageJ (imagej.nih.gov/ij/). 685

For confocal microscopy of AtOFT1 subcellular localization, pollen was harvested as 686

previously described from flowers of 6-week-old oft1-3(-/-)

plants expressing 11p::OFT1-687

GFP or this transgenic line crossed with either Got1p-mCherry or MEMB12-mCherry 688

Golgi marker lines (Geldner et al., 2009) and germinated for 1.5 h. Before imaging with 689

an Olympus FluoView FV1000 line-scanning confocal microscope equipped with 40× 690

1.3 NA and 60× 1.4 NA oil objectives, 488, and 543 nm excitation laser lines (GFP 691

emission filter 500-530; TRITC emission filter 555-615), liquid PGM was applied to the 692

surface of the agar media pad and a coverslip was added. For co-localization imaging 693

using Mitotracker staining, the 11p::OFT1-GFP pollen tubes were treated with 500 nM 694

Mitotracker Orange for 15 min prior to coverslip addition and imaging. Mitotracker 695

Orange was visualized using the TRITC filter described above. Images were processed 696



31

in Fiji (https://fiji.sc/). Co-localization statistics were calculated using JACoP (Bolte and 697

Cordelieres, 2006). 698

699

Pollen tube penetration assays 700

Pollen from 6-week old plants was collected from Col-0 or homozygous oft1 701

mutant lines and used to pollinate mature, emasculated ms1 stigmas. Twenty min after 702

pollination, stigmas were dissected from the parent plant using a razor blade and 703

transferred to a PGM-agarose pad on a microscope slide as described above. Samples 704

were incubated at 25°C in a humidified chamber in the dark for 2 HAP. Pollen tube 705

emergence from the transmitting tract was observed over time. Dissected stigmas were 706

visualized using a Leica EZ4HD dissecting scope at 35X magnification over the indicated 707

time intervals starting 2 HAP. After each set of images was collected for a given time 708

point, the samples were returned to the humidified chamber in the dark until the next set 709

of images was collected. 710

Semi-in vivo assays utilizing fluorescently labeled pollen tubes were assembled 711

identically. Mature pollen was collected from 11p::OFT1-GFP +/-

; oft1-3-/-

or 9p::YFP+/-

712

transgenic lines and used to pollinate mature, emasculated ms1 pistils, which were then 713

dissected and incubated as described above. Four HAP, pollen tubes emerging from the 714

style were examined using a Keyence BZ-X700 microscope at 20X magnification. 715

Sequential Z-stack images using brightfield illumination, GFP (ex. = 470 nm, em. = 525 716

nm), or YFP (ex. = 500 nm, em. = 530 nm) were used to construct images. The Z-stacks 717

were assembled and processed in Keyence BZ-X Analyzer software and subsequently 718



32

analyzed for total pollen tubes emerging from the style as well as the proportion of 719

fluorescent and non-fluorescent tubes. 720

721

Analine Blue Staining of pollinated pistils 722

Mature, emasculated ms1 flowers were pollinated with either Col-0 or oft1-3(-/-)

mutant 723

pollen and incubated under normal growth conditions for 24 h. Following incubation, 724

pistils were dissected away from the remainder of the plant and subjected to analine blue 725

staining (Mori et al., 2006). Images were acquired at 20X with a Keyence BZ-X710 726

epifluorescent microscope fitted with a DAPI filter cube. 727

728

RNA Isolation, cDNA Synthesis, and RT-PCR 729

RNA from 7-day-old seedlings from each homozygous oft1 mutant line and Col-0 was 730

isolated using the PureLink Plant RNA Kit (ThermFisher Scientific) following the 731

manufacturer’s instructions. Genomic DNA contamination was eliminated from the 732

extracts using the Turbo DNA-free Kit (Ambion), and synthesis of first-strand cDNA was 733

carried out using the Invitrogen SuperScript III First-Strand Synthesis System both 734

according to the manufacturer’s protocol (ThermoFisher Scientific). The resulting cDNA 735

library for each line was probed for AtOFT1 transcript, as well as ACTIN7 (ACT7; 736

At1g33160) for amplification reference by PCR. Reactions were assembled using 24 ng 737

cDNA as template and the respective gene-specific primers (OFT1-pENTR F and OFT1-738

pENTR R or ACTIN7 F and ACTIN7 R; Supplemental Table S1). Reactions probing for 739

the AtOFT1 transcript were cycled under the following conditions: 98°C initial 740



33

denaturation for 5 min, followed by 30 cycles of 98°C (30 sec), 55°C (30 sec), 72°C (1 741

min), final elongation for 5 min at 72°C. Reactions probing for the ACTIN7 transcript 742

were cycled under the following conditions: 98°C initial denaturation for 30 sec, 743

followed by 30 cycles of 98°C (10 sec), 55°C (20 sec), 72°C (30 sec), final elongation for 744

5 min at 72°C. The resulting PCR products were separated on 1.0% (w/v) agarose gels 745

and visualized with a Bio-Rad Gel Doc XR+ Image analysis workstation. 746

Accession Numbers 747

AtOFT1 (At3g05320), FRIABLE (FRB1; At5g01100), ESMERELDA1 (ESMD1; 748

At2g01480), MSR1 (At3g21190), MSR2 (At1g51630), SPINDLY (SPY; At3g11540), 749

Homo sapiens POFT1 (NP_056167.1), Mus musculus POFT1 (NP_536711.3), Danio 750

rerio POFT1 (NP_991281.3), Caenorhabditis elegans POFT1 (ABA29469.1), 751

Drosophila melanogaster POFT1 (AAF58290.1). 752

Supplemental Data 753

The following supplemental materials are available. 754

Supplemental Figure S1. PCR genotyping verification of oft1 T-DNA insertions and 755

AtOFT1 transcript abundance. 756

Supplemental Figure S2. Seed set measurements of 11p::OFT1-GFP complement lines. 757

Supplemental Figure S3. Tissue expression analysis of AtOFT1. 758

Supplemental Figure S4. In vitro pollen germination and elongation rates of oft1-3 759

rescued pollen. 760

Supplemental Figure S5. Pollen tube behavior following emergence from SIV pistils. 761



34

Supplemental Figure S6. Decapitation assay experimental design. 762

Supplemental Figure S7. Expression verification of AtOFT1 site-directed mutant 763

constructs. 764

Supplemental Figure S8. Sequence and relevant features of the 11p::OFT1-GFP 765

complementation construct. 766

Supplemental Table S1. Oligonucleotide primers used in this study. 767

Supplemental Movie 1. AtOFT1-GFP subcellular dynamics in growing pollen tubes. 768

769

Acknowledgements 770

DKS, DMJ, JL, and IW were supported by startup funds through the University of 771

Nevada, Reno Department of Biochemistry and Molecular Biology. DMJ and IW were 772

supported by National Science Foundation IOS award 1449068. DKS was also supported 773

by a National Science Foundation Graduate Research Fellowship and a Nevada 774

Agricultural Experiment Station Award (NEV00382). Additionally, the confocal 775

microscope used in this study is supported by an NIH COBRE award (Award number 776

RR024210). 777

778

779

780

781

782

783

784

785

786



35

Tables 787

Table I. Segregation distortion analysis of oft1 mutant and complemented lines. 788

Parents (♂ x ♀)a #Rb #Sb Total %R TEc χ2d P value

oft1-1+/-

self 258 247 505 51.1 1.04 38.5 <0.00001

oft1-2+/-

self 51 57 108 47.2 0.89 11.1 0.0009

oft1-3+/-

self 369 328 697 52.9 1.13 45.2 <0.00001

Col-0 x oft1-1+/-

57 52 109 52.3 1.10 0.11 0.74

oft1-1+/-

x Col-0 0 116 116 0 0 58 <0.00001

Col-0 x oft1-3+/-

364 340 704 51.7 1.07 0.41 0.52

oft1-3+/-

x Col-0 1 1871 1872 0.05 5.3 x10-4 934 <0.00001

Col-0 x oft1-3-/-

; 11p::OFT1-GFP+/-e

84 102 186 45.2 0.82 0.87 0.35

oft1-3-/-

; 11p::OFT1-GFP+/- x oft1-3-/-e

494 0 494 0 ND 247 <0.00001

Col-0 x oft1-3+/-

; 11p::OFT1-GFP+/-e

161 158 319 50.5 1.02 0.014 0.91

oft1-3+/-

; 11p::OFT1-GFP+/- x Col-0e 771 1688 2459 31.4 0.46 171.0 <0.00001

789 a The parent lines of each cross are indicated. Transgenic lines containing the 11p::OFT1-GFP 790 construct are abbreviated as 11p::OFT1-GFP with the indicated genotype. 791 b The total number of resistant and sensitive individuals in each cross F1 progeny are indicated. 792 Transmission of alleles was scored as described in Materials and Methods: oft1-1 (KanR), oft1-2 793 (PCR genotyping), oft1-3 (BarR), 11p::OFT1-GFP (HygR) 794 c Transmission efficiency was calculated as #R/ #S. 795 d χ2 was calculated based on the expectation of a 1:1 segregation of transgenes in the resulting F1 796 populations 797 e The aggregate numbers of F1 progeny from at least three independent complemented lines is 798 represented in the table 799

800

801

802

803

804

805

806

807

Figure legends 808

809

Figure 1. Phylogenetic analysis of the Arabidopsis putative POFT family. A Neighbor-810



36

Joining phylogenetic tree was constructed based on the amino acid sequences of members 811

of the putative Arabidopsis protein O-fucosyltransferase family as well as various 812

metazoan POFT1 sequences. Labels at nodes represent bootstrap values based on 1000 813

bootstrap trials. The clade containing metazoan POFT1 sequences and related 814

Arabidopsis putative POFTs are colored in red. 815

816

Figure 2. Phenotypic characterization of oft1 mutant lines. A, The inflorescence 817

morphology of 6-week-old oft1 mutants and wild-type Col-0 controls. Blue arrowheads 818

indicate improperly developed siliques. Scale bars represent 1 cm. B, Representative 819

images of fully developed siliques harvested from 6-week-old oft1 mutant or wild-type 820

Col-0 control plants. The genotypes of siliques are indicated in the top right corner of 821

each panel. Scale bars represent 2 mm. C, Quantification of 6-week-old Col-0 (black 822

bar) and oft1 mutant (red bars) silique lengths. Data are means ± SEM (n = 23). One-823

way ANOVA analysis indicated a significant difference between Col-0 and oft1 mutants 824

(** indicates P < 0.01 by Tukey’s posthoc analysis). D, Fully developed siliques of the 825

indicated genotypes were harvested from 6-week-old plants and cleared in 70% EtOH as 826

described in Materials and Methods. Representative siliques from each genotype are 827

shown. E, Quantification of Col-0 (black bar) and oft1 mutants (red bars) seed set. Data 828

are means ± SEM (n = 12–22 siliques). One-way ANOVA analysis indicated a 829

significant difference between Col-0 and oft1 mutants (** indicates P < 0.01 by Tukey’s 830

post-hoc analysis). 831

832

Figure 3. Sub-cellular localization of AtOFT1. A, Pollen tubes from 11p::OFT1-GFP-833

expressing plants stained with 500 nM Mitotracker Orange for 15 min (top row panels) 834

and 11p::OFT1-GFP and MEMB12- or GOT1-mCherry (Geldner et al., 2009) co-835

expressing plants (middle and bottom row panels, respectively). Pollen tubes were 836

germinated as described above and visualized by confocal microscopy. The outline of 837

each pollen tube is shown as a dashed white line. OFT1-GFP (left column panels; green 838

signal) and co-localization marker (middle column panels; magenta signal) and the merge 839

of each image set is shown (right column panels; white signal). Scale bars in all images 840

represent 10 µm. B, Quantitative co-localization analysis of each image set was 841

performed using JACoP (Bolte and Cordelieres, 2006), and the Pearson Correlation 842

Coefficient between OFT1-GFP and MitoTracker (black bar), MEMB12-mCherry 843

(charcoal bar), or Got1p-mCherry (gray bar) was calculated. Data are means ± SEM (n = 844

6–15 independent images per co-localization marker). C, Live-cell confocal imaging of 845

OFT1-GFP subcellular localization in a growing pollen tube over time. The outline of 846



37

the pollen tube is indicated with a dashed white line. Image timepoints are indicated in 847

the upper left corner of each image. The trajectories of three representative particles are 848

individually indicated at their current position for each indicated time point by green, red, 849

and yellow arrowheads and their corresponding trajectories in successive images 850

indicated with green, red, and yellow lines, respectively. 851

852

853

Figure 4. In vitro pollen tube growth behavior of oft1 mutants. A, Representative 854

images of wild-type Col-0 or oft1 mutant pollen are shown after 6 h of growth. The 855

pollen tube genotype is indicated at the top left corner of each image. Scale bars 856

represent 100 µm. B, Lengths of Col-0 (black line), oft1-1 (red line), oft1-2 (blue line), 857

and oft1-3 (green line) pollen tubes were quantified at the indicated time point. Data are 858

means ± SEM (n = 30–240). 859

860

861

Figure 5. Pollen tube penetration behavior of oft1 mutants. A, Analine blue staining of 862

ms1 pistils pollinated with either Col-0 (top panel) or oft1-1 (bottom panel) pollen. Scale 863

bars represent 300 µm. White arrowheads indicate pollen tube trajectories through the 864

pistil. B, Representative images of pollen tube emergence from the transmitting tract in 865

ms1 pistils dissected at the indicated timepoints following pollination with either wild-866

type Col-0 (top row panels) or oft1-1 (bottom row panels) pollen. Scale bars represent 867

0.5 mm. C, Quantification of ms1 stigmas exhibiting pollen tubes emanating from the 868

transmitting tract following pollination with Col-0 (black bars), oft1-1 (red bars), oft1-2 869

(blue bars), or oft1-3 (green bars) homozygous pollen. Data are means ± SEM (n = 4 870

independent trials with 4–8 stigmas per experimental group per trial). D, Quantification 871

of pollen tube number emerging from ms1 stigmas at the indicated time points following 872

pollination with Col-0 (black line), oft1-1 (red line), oft1-2 (blue line), or oft1-3 (green 873

line). Data are means ± SEM (n = 4 biological replicates). 874

875

876

Figure 6. Semi-in vivo penetration competition assay of oft1-3-/-

; 11p::OFT1-GFP+/-

877

pollen tubes. A, GFP and brightfield overlaid images depicting the bottom of an ms1 878

stigma displaying penetrating pollen tubes. Red arrowheads indicate non-fluorescent 879

pollen tubes and yellow arrowheads indicate fluorescent pollen tubes. B, Quantification 880

of the percentage of fluorescent pollen tubes emerging from the transmitting tract 4 h 881

after pollination. Three independent oft1-3-/-

;11p:OFT1-GFP+/-

transgenic lines (red bars) 882

were assessed. 9p::YFP+/-

pollen tubes (black bar) served as a negative control. Data are 883



38

means ± SD (n ≥ 7). 884

885

Figure 7. Structure-function analysis of AtOFT1 putative catalytic residues. A, The 886

active site of C. elegans POFT1 bound to GDP-Fucose (PDB ID 3ZY5) is shown with 887

critical catalytic residue indices in black font, and the corresponding AtOFT1 residues 888

shown in blue font. B, Sequence alignments of the AtOFT1 N-terminal (upper panel) and 889

C-terminal (lower panel) regions compared to multiple metazoan POFT1s. Critical 890

catalytic residues are highlighted in red, and the beginning residues for each alignment 891

segment are indicated for reference. An aspartic acid residue used to propose an 892

alternative alignment of the N-terminal domain is boxed in blue. C, Seed set in oft1-1 893

plants expressing the indicated site-directed mutant constructs of AtOFT1 (black 894

bars). Seed sets of Col-0 (gray bar), oft1-1 (red bar), oft1-2 (blue bar), and oft1-1 lines 895

expressing wild-type AtOFT1 (green bars) are shown for reference. Data are means ± 896

SEM (n = 29–71). 897

898

899

900

901

Literature cited 902

Alonso, J. M., Stepanova, A. N., Leisse, T. J., Kim, C. J., Chen, H., Shinn, P., et al. 903

(2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 904

653-7. 905

906

Beale, K. M., and Johnson, M. A. (2013) Speed dating, rejection, and finding the perfect 907

mate: advice from flowering plants. Curr. Opin. Plant Biol. 16: 590-7. 908

909

Boavida, L. C., and McCormick, S. (2007) Temperature as a determinant factor for 910

increased and reproducible in vitro pollen germination in Arabidopsis thaliana. Plant J. 911

52: 570-82. 912

913

Bolte, S., and Cordelieres, F. P. (2006) A guided tour into subcellular colocalization 914

analysis in light microscopy. J. Microsc. 224: 213-32. 915

916

Bouton, S., Leboeuf, E., Mouille, G., Leydecker, M. T., Talbotec, J., Granier, F., Lahaye, 917

M., Hofte, H., and Troung, H. N. (2002) QUASIMODO1 encodes a putative membrane-918

bound glycosyltransferase required for normal pectin synthesis and cell adhesion in 919

Arabidopsis. Plant Cell 14: 2477-90. 920

921



39

Chae, K., and Lord, E. M. (2011) Pollen tube growth and guidance: roles of small, 922

secreted proteins. Ann. Bot. 108: 627-36. 923

924

Clough, S. J., and Bent, A. F. (1998) Floral dip: a simplified method for Agrobacterium-925

mediated transformation of Arabidopsis thaliana. Plant J. 16: 735-43. 926

927

Durand, C., Vicre-Gibouin, M., Follet-Gueye, M. L., Duponchel, L., Moreau, M., 928

Lerouge, P., and Driouich, A. (2009) The organization pattern of root border-like cells 929

of Arabidopsis is dependent on cell wall homogalacturonan. Plant Physiol. 150: 1411-21. 930

931

Elleman, C. J., Franklin-Tong, V., Dickinson, H. G. (1992) Pollination in species with 932

dry stigmas: the nature of the early stigmatic response and the pathway taken by pollen 933

tubes. New Phytol. 121: 413-24. 934

935

Edwards, K., Johnstone, C., and Thompson, C. (1991) A simple and rapid method for 936

the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 19: 1349. 937

938

Frietsch, S., Wang, Y. F., Sladek, C., Poulsen, L. R., Romanowsky, S. M., Schroeder, J. 939

I., and Harper, J. F. (2007) A cyclic nucleotide-gated channel is essential for polarized 940

tip growth of pollen. PNAS 104: 14531-6. 941

942

Geldner, N., Denervaud-Tendon, V., Hyman, D. L., Mayer, U., Stierhof, Y. D., and 943

Chory, J. (2009) Rapid, combinatorial analysis of membrane compartments in intact 944

plants with a multicolor marker set. Plant J. 59: 169-78. 945

946

Goldman, M. H. S., Goldberg, R. B., and Mariani, C. (1994) Female sterile tobacco 947

plants are produced by stigma-specific cell ablation. EMBO J. 13: 2976-84. 948

949

Grefen, C., Donald, N., Hashimoto, K., Kudla, J., Schumacher, K., and Blatt, M. R. 950

(2010) A ubiquitin-10 promoter-based vector set for fluorescent protein tagging 951

facilitates temporal stability and native protein distribution in transient and stable 952

expression studies. Plant J. 64: 355-65. 953

954

Hao, L., Liu, J., Zhong, S., Gu, H., and Qu, L. J. (2016) AtVPS41-mediated endocytic 955

pathway is essential for pollen tube-stigma interaction in Arabidopsis. PNAS 113: 6307-956

12. 957

958

Harris, R. J., and Spellman, M. W. (1993) O-linked fucose and other post-translation 959

modifications unique to EGF module. Glycobiology 3: 219-24. 960

961

Hellens, R. P., Edwards, E. A., Leyland, N. R., Bean, S., and Mullineaux, P. M. (2000) 962

pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant 963

transformation. Plant Mol. Biol. 42: 819-32. 964

965



40

Jiang, L., Yang, S. L., Xie, L. F., Puah, C. S., Zhang, X. Q., Yang, W. C., Sundarsen, V., 966

Ye, D. (2005) VANGUARD1 encodes a pectin methylesterase that enhances pollen tube 967

growth in the Arabidopsis style and transmitting tract. Plant Cell 17: 584-96. 968

969

Kohorn, B. D. (2016) Cell wall-associated kinases and pectin perception. J. Exp. Bot. 970

67: 489-94. 971

972

Kovall, R. A., Gebelein, B., Sprinzak, D., and Kopan, R. (2017) The canonical Notch 973

signaling pathway: Structure and biochemical insights into shape, sugar, and force. Dev. 974

Cell. 41: 228-241. 975

976

Leonhard-Melief, C., and Haltiwanger, R. S. (2010) O-fucosylation of thrombospondin 977

type 1 repeats. Methods Enzymol. 480: 401-16. 978

979

Levitin, B., Richter, D., Markovich, I., and Zik, M. (2008) Arabinogalactan proteins 6 980

and 11 are required for stamen and pollen function in Arabidopsis. Plant J. 56: 351-63. 981

982

Leydon, A. R., Tsukamoto, T., Dunatunga, D., Qin, Y., Johnson, M. A., Palanivelu, R. 983

(2015) Pollen tube discharge completes the process of synergid degeneration that is 984

initiated by pollen tube-synergid interaction in Arabidopsis. Plant Physiol. 169: 485-96. 985

986

Lira-Navarette, E., Valero-Gonzalez, J., Villanueva, R., Martinez-Julvez, M., Tejero, T., 987

Merino, P., Panjikar, S., and Hurtado-Guerrero, R. (2011) Structural insights into the 988

mechanism of protein O-fucosylation. PLoS ONE 6: e25365. 989

990

Luca, V. C., Kim, B. C., Ge, C., Kakuda, S., Wu, D., Roein-Peikar, M., et al. (2017) 991

Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity. 992

Science 355: 1320-24. 993

994

Luo, Y., and Haltiwanger, R. S. (2005) O-fucosylation of notch occurs in the 995

endoplasmic reticulum. J. Biol. Chem. 280: 11289-94. 996

997

Mizukami, A. G., Inatsugi, R., Jiao, J., Kotake, T., Kuwata, K., Ootani, K., et al. (2016) 998

The AMOR arabinogalactan sugar chain induces pollen-tube competency to respond to 999

ovular guidance. Curr. Biol. 26: 1091-7. 1000

1001

Mori T., Kuroiwa H., Higasiyama T., and Kuroiwa T. (2006). Generative Cell Specific 1 1002

is essential for angiosperm fertilization. Nature Cell Biology 8: 64-71. 1003

1004

Myers, C., Romanowsky, S. M., Barron, Y. D., Garg, S., Azuse, C. L., Curran, A., Davis, 1005

R. M., Hatton, J., Harmon, A. C., and Harper, J. F. (2009) Calcium-dependent protein 1006

kinases regulate polarized tip growth in pollen tubes. Plant J. 59: 528-39 1007

1008

Nakagawa, T., Suzuki, T., Murata, S., Nakamura, S., Hino, T., Maeo, K., Tabata, R., 1009

Kawai, T., Tanaka, K., Niwa, Y., Watanabe, Y., Nakamura, K., Kimura, T., and Ishiguro, 1010

S. (2007) Improved gateway binary vectors: high-performance vectors for creation of 1011



41

fusion constructs in transgenic analysis of plants. Biosci. Biotechnol. Biochem. 71: 1012

2095-100. 1013

1014

Neumetzler, L., Humphrey, T., Lumba, S., Snyder, S., Yeats, T. H., Usadel, B., 1015

Vasilevski, A., Patel, J., Rose, J. K., Persson, S., and Bonetta, D. (2012) The 1016

FRIABLE1 gene product affects cell adhesion in Arabidopsis. PLoS ONE 7: e42914. 1017

1018

Okajima, T., Xu, A., and Irvine, K. D. (2003) Modulation of notch-ligand binding by 1019

protein O-fucosyltransferase 1 and fringe. J. Biol. Chem. 278: 42340-5. 1020

1021

Okuda, S., Tsutsui, H., Shiina, K., Sprunck, S., Takeuchi, H., Yui, R., et al. (2009) 1022

Defensin-like polypeptide LUREs are pollen tube attractants secreted from the synergid 1023

cells. Nature 458: 357-61. 1024

1025

Palanivelu, R., and Johnson, M. A. (2010) Functional genomics of pollen tube-pistil 1026

interactions in Arabidopsis. Biochem. Soc. Trans. 38: 593-7. 1027

1028

Palanivelu, R., Brass, L., Edlund, A. F., Preuss, D. (2003) Pollen tube growth and 1029

guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 1030

114: 47-59. 1031

1032

Qin, Y., Leydon, A. R., Manziello, A., Pandey, R., Mount, D., Denic, S., Vasic, B., 1033

Johnson, M. A., and Palanivelu, R. (2009) Penetration of the stigma and style elicits a 1034

novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil. PLoS 1035

Genet. 5: 1000621. 1036

1037

Rampal, R., Li, A. S., Moloney, D. J., Georgiou, S. A., Luther, K. B., Nita-Lazar, A., and 1038

Haltiwanger, R. S. (2005) Lunatic fringe, maniac fringe, and radical fringe recognize 1039

similar specificity determinants in O-fucosylated epidermal growth factor-like repeats. J. 1040

Biol. Chem. 280: 42454-63. 1041

1042

Rebay, I., Fleming, R. J., Fehon, R. G., Cherbas, L., Cherbas, P., and Artavanis-1043

Tsakonas, S. (1991) Specific EGF repeats of Notch mediate interactions with Delta and 1044

Serrate: implications for Notch as a multifunctional receptor. Cell 67: 687-99. 1045

1046

Sandaklie-Nikolova, L., Palanivelu, R., King, E. J., Copenhavaer, G. P., and Drews, G. 1047

N. (2007) Synergid cell death in Arabidopsis is triggered following direct interaction 1048

with the pollen tube. Plant Physiol. 144: 1753-62. 1049

1050

Sasamura, T., Sasaki, N., Miyashita, F., Nakao, S., Ishikawa, H. O., Ito, M., Kitagawa, 1051

M., Harigaya, K., Spana, E., Bilder, D., Perrimon, N., and Matsuno, K. (2003) Neurotic, 1052

a novel maternal neurogenic gene, encodes an O-fucosyltransferase that is essential for 1053

Notch-Delta interactions. Development 130: 4785-95. 1054

1055



42

Schiott, M., Romanowsky, S. M., Baekgaard, L., Jakobsen, M. K., Palmgren, M. G., and 1056

Harper, J. F. (2004) A plant plasma membrane calcium pump is required for normal 1057

pollen tube growth and fertilization. PNAS 101: 9502-7. 1058

1059

Sessions, R. A., Zambryski, P. C. (1995) Arabidopsis gynoecium structure in the wild 1060

and in ettin mutants. Development 121: 1519-32. 1061

1062

Shi, S., and Stanley, P. (2003) Protein O-fucosyltransferase 1 is an essential component 1063

of Notch signaling pathways. PNAS 100: 5234-9. 1064

1065

Stahl, M., Uemura, K., Ge, C., Shi, S., Tashima, Y., and Stanley, P. (2008) Roles of 1066

Pofut1 and O-fucose in mammalian Notch signaling. J. Biol. Chem. 283: 13638-51. 1067

1068

Takeuchi, H., Higashiyama, T. (2012) A species-specific cluster of defensing-like genes 1069

encodes diffusible pollen tube attractants in Arabidopsis. PLoS Biol. 10: e1001449. 1070

1071

Takeuchi, H., Higashiyama, T. (2016) Tip-localized receptors control pollen tube 1072

growth and LURE sensing in Arabidopsis. Nature 531: 245-8. 1073

1074

Tanz, S. K., Castleden, I., Hooper, C. M., Vacher, M., Small, I., and Millar, H. A. (2013) 1075

SUBA3: a database for integrating experimentation and prediction to define the 1076

SUBcellular location of proteins in Arabidopsis. Nucleic Acids Res. 41: D1185-91. 1077

1078

Trindade, H., Boavida, L. C., Borralho, N., and Fiejo, J. A. (2001) Successful 1079

fertilization and seed set from pollination on immature non-dehisced flowers of 1080

Eucalyptus globulus. Ann. Bot. 87: 469-475. 1081

1082

van Tuyl, J. M., van Dien, M. P., van Creij, M. G. M., van Kleinwee, T. C. M., Franken, 1083

J., and Bino, R. J. (1991) Application of in vitro pollination, ovary cultures, ovule 1084

culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium 1085

crosses. Plant Sci. 74: 115-26. 1086

1087

Verger, S., Chabout, S., Gineau, E., and Mouille, G. (2016) Cell adhesion in plant is 1088

under the control of putative O-fucosyltransferases. Development 143: 2536-40. 1089

1090

Villalobos, J. A., Yi, B. R., and Wallace, I. S. (2015) 2-fluoro-L-fucose is a 1091

metabolically incorporated inhibitor of plant cell wall polysaccharide biosynthesis. PLoS 1092

ONE 10: e0139091 1093

1094

Wang, Y., Shao, L., Shi, S., Harris, R. J., Spellman, M. W., Stanley, P., and Haltiwanger, 1095

R. S. (2001) Modification of epidermal growth factor-like repeats with O-fucose: 1096

Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase. 1097

J. Biol. Chem. 276: 40338-45. 1098

1099



43

Wang, Y., Mortimer, J. C., Davis, J., Dupree, P., and Keegstra, K. (2013) Identification 1100

of an additional protein involved in mannan biosynthesis. Plant J. 73: 105-17. 1101

1102

Wilson, Z. A., Morroll, S. M., Dawson, J., Swarup, R., and Tighe, P. J. (2001) The 1103

Arabidopsis MALE STERILITY (MS1) gene is a transcriptional regulator of male 1104

gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J. 1105

28: 27-39. 1106

1107

Zentella, R., Sui, N., Barnhill, B., Hsieh, W. P., Hu, J., Shabanowitz, J., et al. (2017) 1108

The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor 1109

DELLA. Nat. Chem. Biol. 13: 479-85. 1110

1111

1112



Figure 1. Phylogenetic analysis of the Arabidopsis putative POFT family. A Neighbor-Joining phylogenetic

tree was constructed based on the amino acid sequences of members of the putative Arabidopsis protein O-

fucosyltransferase family as well as various metazoan POFT1 sequences. Labels at nodes represent bootstrap

values based on 1000 bootstrap trials. The clade containing metazoan POFT1 sequences and related

Arabidopsis putative POFTs are colored in red.

Smith et al., Figure 1



Col-0 oft1-1 oft1-2 oft1-3


Figure 2. Phenotypic characterization of oft1 mutant lines. A, The inflorescence morphology of 6-week-old

oft1 mutants and wild-type Col-0 controls. Blue arrowheads indicate improperly developed siliques. Scale

bars represent 1 cm. B, Representative images of fully developed siliques harvested from 6-week-old oft1

mutant or wild-type Col-0 control plants. The genotypes of siliques are indicated in the top right corner of

each panel. Scale bars represent 2 mm. C, Quantification of 6-week-old Col-0 (black bar) and oft1 mutant

(red bars) silique lengths. Data are means ± SEM (n = 23). One-way ANOVA analysis indicated a significant

difference between Col-0 and oft1 mutants (** indicates P < 0.01 by Tukey’s posthoc analysis). D, Fully

developed siliques of the indicated genotypes were harvested from 6-week-old plants and cleared in 70%

EtOH as described in Materials and Methods. Representative siliques from each genotype are

shown. E, Quantification of Col-0 (black bar) and oft1 mutants (red bars) seed set. Data are means ± SEM (n

= 12–22 siliques). One-way ANOVA analysis indicated a significant difference between Col-0 and oft1

mutants (** indicates P < 0.01 by Tukey’s post-hoc analysis).

Col-0 oft1-1

oft1-3oft1-2

Col-0 oft1-1

oft1-2 oft1-3

A

D

C

E

B



0s 2s

4s 9s

OFT1-GFP

OFT1-GFP

OFT1-GFP

MitoTracker

MEMB12-mCherry

Got1p-mCherry

Merge

Merge

Merge

A

CB

OFT1-GFP

MitoTracker

OFT1-GFP

MEMB12

OFT1-GFP

Got1p


Figure 3. Sub-cellular localization of AtOFT1. A, Pollen tubes from 11p::OFT1-GFP-expressing plants

stained with 500 nM Mitotracker Orange for 15 min (top row panels) and 11p::OFT1-GFP and MEMB12- or

GOT1-mCherry (Geldner et al., 2009) co-expressing plants (middle and bottom row panels,

respectively). Pollen tubes were germinated as described above and visualized by confocal microscopy. The

outline of each pollen tube is shown as a dashed white line. OFT1-GFP (left column panels; green signal) and

co-localization marker (middle column panels; magenta signal) and the merge of each image set is shown

(right column panels; white signal). Scale bars in all images represent 10 µm. B, Quantitative co-localization

analysis of each image set was performed using JACoP (Bolte and Cordelieres, 2006), and the Pearson

Correlation Coefficient between OFT1-GFP and MitoTracker (black bar), MEMB12-mCherry (charcoal bar),

or Got1p-mCherry (gray bar) was calculated. Data are means ± SEM (n = 6–15 independent images per co-

localization marker). C, Live-cell confocal imaging of OFT1-GFP subcellular localization in a growing pollen

tube over time. The outline of the pollen tube is indicated with a dashed white line. Image timepoints are

indicated in the upper left corner of each image. The trajectories of three representative particles are

individually indicated at their current position for each indicated time point by green, red, and yellow

arrowheads and their corresponding trajectories in successive images indicated with green, red, and yellow

lines, respectively.https://plantphysiol.orgDownloaded on February 4, 2021. - Published by

Copyright (c) 2020 American Society of Plant Biologists. All rights reserved.


Col-0 oft1-1

oft1-2 oft1-3

Figure 4. In vitro pollen tube growth behavior of oft1 mutants. A, Representative images of wild-type Col-0

or oft1 mutant pollen are shown after 6 h of growth. The pollen tube genotype is indicated at the top left

corner of each image. Scale bars represent 100 µm. B, Lengths of Col-0 (black line), oft1-1 (red line), oft1-2

(blue line), and oft1-3 (green line) pollen tubes were quantified at the indicated time point. Data are means ±

SEM (n = 30–240).

A


B

Time (h)



2 h 8 h4 h3 h

Co

l-0

oft

1-1

B

Figure 5. Pollen tube penetration behavior of oft1 mutants. A, Analine blue staining of ms1 pistils pollinated

with either Col-0 (top panel) or oft1-1 (bottom panel) pollen. Scale bars represent 300 µm. White arrowheads

indicate pollen tube trajectories through the pistil. B, Representative images of pollen tube emergence from

the transmitting tract in ms1 pistils dissected at the indicated timepoints following pollination with either wild-

type Col-0 (top row panels) or oft1-1 (bottom row panels) pollen. Scale bars represent 0.5

mm. C, Quantification of ms1 stigmas exhibiting pollen tubes emanating from the transmitting tract following

pollination with Col-0 (black bars), oft1-1 (red bars), oft1-2 (blue bars), or oft1-3 (green bars) homozygous

pollen. Data are means ± SEM (n = 4 independent trials with 4–8 stigmas per experimental group per

trial). D, Quantification of pollen tube number emerging from ms1 stigmas at the indicated time points

following pollination with Col-0 (black line), oft1-1 (red line), oft1-2 (blue line), or oft1-3 (green line). Data

are means ± SEM (n = 4 biological replicates).


C

A

Col-0

oft1-1

D

Time (h) Time (h)



Figure 6. Semi-in vivo penetration competition assay of oft1-3-/-; 11p::OFT1-GFP+/- pollen tubes. A, GFP

and brightfield overlaid images depicting the bottom of an ms1 stigma displaying penetrating pollen tubes.

Red arrowheads indicate non-fluorescent pollen tubes and yellow arrowheads indicate fluorescent pollen

tubes. B, Quantification of the percentage of fluorescent pollen tubes emerging from the transmitting tract 4

h after pollination. Three independent oft1-3-/-;11p:OFT1-GFP+/- transgenic lines (red bars) were

assessed. 9p::YFP+/- pollen tubes (black bar) served as a negative control. Data are means ± SD (n ≥ 7).


A

B




H54N

H54A

R51A

R260AR260K D264A

WT

MmPOFT1 43 CPCMGRFGNQADHFLG

HsPOFT1 38 CPCMGRFGNQADHFLG

DrPOFT1 43 CPCMGRFGNQVDHFLG

DmPOFT1 36 CPCMGRFGNQADHFLG

CePOFT1 35 CPCMGRFGNQVDQFLG

AtOFT1 47 SLLF~RDRHMSDSSST

R40

R51

D244

D264

N43

H54

R240

R260

MmPOFT1 239 YVGIHLRIGSDWKN

HsPOFT1 234 YVGIHLRIGSDWKN

DrPOFT1 239 YVGIHLRIGSDWQN

DmPOFT1 239 FLGIHLRNGIDWVR

CePOFT1 234 FVAVHLRNDADWVR

AtOFT1 254 FVAVHMRIEIDWMI

Figure 7. Structure-function analysis of AtOFT1 putative catalytic residues. A, The active site of C.

elegans POFT1 bound to GDP-Fucose (PDB ID 3ZY5) is shown with critical catalytic residue indices in

black font, and the corresponding AtOFT1 residues shown in blue font. B, Sequence alignments of the

AtOFT1 N-terminal (upper panel) and C-terminal (lower panel) regions compared to multiple metazoan

POFT1s. Critical catalytic residues are highlighted in red, and the beginning residues for each alignment

segment are indicated for reference. An aspartic acid residue used to propose an alternative alignment of the

N-terminal domain is boxed in blue. C, Seed set in oft1-1 plants expressing the indicated site-directed

mutant constructs of AtOFT1 (black bars). Seed sets of Col-0 (gray bar), oft1-1 (red bar), oft1-2 (blue bar),

and oft1-1 lines expressing wild-type AtOFT1 (green bars) are shown for reference. Data are means ± SEM

(n = 29–71).

A B

C



Parsed CitationsAlonso, J. M., Stepanova, A. N., Leisse, T. J., Kim, C. J., Chen, H., Shinn, P., et al. (2003) Genome-wide insertional mutagenesis ofArabidopsis thaliana. Science 301: 653-7.

Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Beale, K. M., and Johnson, M. A. (2013) Speed dating, rejection, and finding the perfect mate: advice from flowering plants. Curr. Opin.Plant Biol. 16: 590-7.


Boavida, L. C., and McCormick, S. (2007) Temperature as a determinant factor for increased and reproducible in vitro pollengermination in Arabidopsis thaliana. Plant J. 52: 570-82.


Bolte, S., and Cordelieres, F. P. (2006) A guided tour into subcellular colocalization analysis in light microscopy. J. Microsc. 224: 213-32.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Bouton, S., Leboeuf, E., Mouille, G., Leydecker, M. T., Talbotec, J., Granier, F., Lahaye, M., Hofte, H., and Troung, H. N. (2002)QUASIMODO1 encodes a putative membrane-bound glycosyltransferase required for normal pectin synthesis and cell adhesion inArabidopsis. Plant Cell 14: 2477-90.


Chae, K., and Lord, E. M. (2011) Pollen tube growth and guidance: roles of small, secreted proteins. Ann. Bot. 108: 627-36.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Clough, S. J., and Bent, A. F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.Plant J. 16: 735-43.


Durand, C., Vicre-Gibouin, M., Follet-Gueye, M. L., Duponchel, L., Moreau, M., Lerouge, P., and Driouich, A. (2009) The organizationpattern of root border-like cells of Arabidopsis is dependent on cell wall homogalacturonan. Plant Physiol. 150: 1411-21.


Elleman, C. J., Franklin-Tong, V., Dickinson, H. G. (1992) Pollination in species with dry stigmas: the nature of the early stigmaticresponse and the pathway taken by pollen tubes. New Phytol. 121: 413-24.


Edwards, K., Johnstone, C., and Thompson, C. (1991) A simple and rapid method for the preparation of plant genomic DNA for PCRanalysis. Nucleic Acids Res. 19: 1349.


Frietsch, S., Wang, Y. F., Sladek, C., Poulsen, L. R., Romanowsky, S. M., Schroeder, J. I., and Harper, J. F. (2007) A cyclic nucleotide-gated channel is essential for polarized tip growth of pollen. PNAS 104: 14531-6.


Geldner, N., Denervaud-Tendon, V., Hyman, D. L., Mayer, U., Stierhof, Y. D., and Chory, J. (2009) Rapid, combinatorial analysis ofmembrane compartments in intact plants with a multicolor marker set. Plant J. 59: 169-78.


Goldman, M. H. S., Goldberg, R. B., and Mariani, C. (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation.https://plantphysiol.orgDownloaded on February 4, 2021. - Published by


http://www.ncbi.nlm.nih.gov/pubmed?term=Alonso%2C J%2E M%2E%2C Stepanova%2C A%2E N%2E%2C Leisse%2C T%2E J%2E%2C Kim%2C C%2E J%2E%2C Chen%2C H%2E%2C Shinn%2C P%2E%2C et al%2E %282003%29 Genome%2Dwide insertional mutagenesis of Arabidopsis thaliana%2E Science 301%3A 653%2D7%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Alonso, J. M., Stepanova, A. N., Leisse, T. J., Kim, C. J., Chen, H., Shinn, P., et al. (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 653-7.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Alonso,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Genome-wide insertional mutagenesis of Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Genome-wide insertional mutagenesis of Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Alonso,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Beale%2C K%2E M%2E%2C and Johnson%2C M%2E A%2E %282013%29 Speed dating%2C rejection%2C and finding the perfect mate%3A advice from flowering plants%2E Curr%2E Opin%2E Plant Biol%2E 16%3A 590%2D7%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Beale, K. M., and Johnson, M. A. (2013) Speed dating, rejection, and finding the perfect mate: advice from flowering plants. Curr. Opin. Plant Biol. 16: 590-7.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Beale,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Speed dating, rejection, and finding the perfect mate: advice from flowering plants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Speed dating, rejection, and finding the perfect mate: advice from flowering plants&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Beale,&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Boavida%2C L%2E C%2E%2C and McCormick%2C S%2E %282007%29 Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana%2E Plant J%2E 52%3A 570%2D82%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Boavida, L. C., and McCormick, S. (2007) Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana. Plant J. 52: 570-82.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Boavida,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Boavida,&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Bolte%2C S%2E%2C and Cordelieres%2C F%2E P%2E %282006%29 A guided tour into subcellular colocalization analysis in light microscopy%2E J%2E Microsc%2E 224%3A 213%2D32%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Bolte, S., and Cordelieres, F. P. (2006) A guided tour into subcellular colocalization analysis in light microscopy. J. Microsc. 224: 213-32.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Bolte,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A guided tour into subcellular colocalization analysis in light microscopy&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A guided tour into subcellular colocalization analysis in light microscopy&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Bolte,&as_ylo=2006&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Bouton%2C S%2E%2C Leboeuf%2C E%2E%2C Mouille%2C G%2E%2C Leydecker%2C M%2E T%2E%2C Talbotec%2C J%2E%2C Granier%2C F%2E%2C Lahaye%2C M%2E%2C Hofte%2C H%2E%2C and Troung%2C H%2E N%2E %282002%29 QUASIMODO1 encodes a putative membrane%2Dbound glycosyltransferase required for normal pectin synthesis and cell adhesion in Arabidopsis%2E Plant Cell 14%3A 2477%2D90%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Bouton, S., Leboeuf, E., Mouille, G., Leydecker, M. T., Talbotec, J., Granier, F., Lahaye, M., Hofte, H., and Troung, H. N. (2002) QUASIMODO1 encodes a putative membrane-bound glycosyltransferase required for normal pectin synthesis and cell adhesion in Arabidopsis. Plant Cell 14: 2477-90.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Bouton,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=QUASIMODO1 encodes a putative membrane-bound glycosyltransferase required for normal pectin synthesis and cell adhesion in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=QUASIMODO1 encodes a putative membrane-bound glycosyltransferase required for normal pectin synthesis and cell adhesion in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Bouton,&as_ylo=2002&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Chae%2C K%2E%2C and Lord%2C E%2E M%2E %282011%29 Pollen tube growth and guidance%3A roles of small%2C secreted proteins%2E Ann%2E Bot%2E 108%3A 627%2D36%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Chae, K., and Lord, E. M. (2011) Pollen tube growth and guidance: roles of small, secreted proteins. Ann. Bot. 108: 627-36.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Chae,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Pollen tube growth and guidance: roles of small, secreted proteins&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Pollen tube growth and guidance: roles of small, secreted proteins&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Chae,&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Clough%2C S%2E J%2E%2C and Bent%2C A%2E F%2E %281998%29 Floral dip%3A a simplified method for Agrobacterium%2Dmediated transformation of Arabidopsis thaliana%2E Plant J%2E 16%3A 735%2D43%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Clough, S. J., and Bent, A. F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16: 735-43.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Clough,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Clough,&as_ylo=1998&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Durand%2C C%2E%2C Vicre%2DGibouin%2C M%2E%2C Follet%2DGueye%2C M%2E L%2E%2C Duponchel%2C L%2E%2C Moreau%2C M%2E%2C Lerouge%2C P%2E%2C and Driouich%2C A%2E %282009%29 The organization pattern of root border%2Dlike cells of Arabidopsis is dependent on cell wall homogalacturonan%2E Plant Physiol%2E 150%3A 1411%2D21%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Durand, C., Vicre-Gibouin, M., Follet-Gueye, M. L., Duponchel, L., Moreau, M., Lerouge, P., and Driouich, A. (2009) The organization pattern of root border-like cells of Arabidopsis is dependent on cell wall homogalacturonan. Plant Physiol. 150: 1411-21.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Durand,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The organization pattern of root border-like cells of Arabidopsis is dependent on cell wall homogalacturonan&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The organization pattern of root border-like cells of Arabidopsis is dependent on cell wall homogalacturonan&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Durand,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Elleman%2C C%2E J%2E%2C Franklin%2DTong%2C V%2E%2C Dickinson%2C H%2E G%2E %281992%29 Pollination in species with dry stigmas%3A the nature of the early stigmatic response and the pathway taken by pollen tubes%2E New Phytol%2E 121%3A 413%2D24%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Elleman, C. J., Franklin-Tong, V., Dickinson, H. G. (1992) Pollination in species with dry stigmas: the nature of the early stigmatic response and the pathway taken by pollen tubes. New Phytol. 121: 413-24.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Elleman,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Pollination in species with dry stigmas: the nature of the early stigmatic response and the pathway taken by pollen tubes&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Pollination in species with dry stigmas: the nature of the early stigmatic response and the pathway taken by pollen tubes&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Elleman,&as_ylo=1992&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Edwards%2C K%2E%2C Johnstone%2C C%2E%2C and Thompson%2C C%2E %281991%29 A simple and rapid method for the preparation of plant genomic DNA for PCR analysis%2E Nucleic Acids Res%2E 19%3A 1349%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Edwards, K., Johnstone, C., and Thompson, C. (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 19: 1349.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Edwards,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A simple and rapid method for the preparation of plant genomic DNA for PCR analysis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A simple and rapid method for the preparation of plant genomic DNA for PCR analysis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Edwards,&as_ylo=1991&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Frietsch%2C S%2E%2C Wang%2C Y%2E F%2E%2C Sladek%2C C%2E%2C Poulsen%2C L%2E R%2E%2C Romanowsky%2C S%2E M%2E%2C Schroeder%2C J%2E I%2E%2C and Harper%2C J%2E F%2E %282007%29 A cyclic nucleotide%2Dgated channel is essential for polarized tip growth of pollen%2E PNAS 104%3A 14531%2D6%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Frietsch, S., Wang, Y. F., Sladek, C., Poulsen, L. R., Romanowsky, S. M., Schroeder, J. I., and Harper, J. F. (2007) A cyclic nucleotide-gated channel is essential for polarized tip growth of pollen. PNAS 104: 14531-6.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Frietsch,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A cyclic nucleotide-gated channel is essential for polarized tip growth of pollen&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A cyclic nucleotide-gated channel is essential for polarized tip growth of pollen&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Frietsch,&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Geldner%2C N%2E%2C Denervaud%2DTendon%2C V%2E%2C Hyman%2C D%2E L%2E%2C Mayer%2C U%2E%2C Stierhof%2C Y%2E D%2E%2C and Chory%2C J%2E %282009%29 Rapid%2C combinatorial analysis of membrane compartments in intact plants with a multicolor marker set%2E Plant J%2E 59%3A 169%2D78%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Geldner, N., Denervaud-Tendon, V., Hyman, D. L., Mayer, U., Stierhof, Y. D., and Chory, J. (2009) Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. Plant J. 59: 169-78.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Geldner,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Geldner,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1


EMBO J. 13: 2976-84.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Grefen, C., Donald, N., Hashimoto, K., Kudla, J., Schumacher, K., and Blatt, M. R. (2010) A ubiquitin-10 promoter-based vector set forfluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies.Plant J. 64: 355-65.


Hao, L., Liu, J., Zhong, S., Gu, H., and Qu, L. J. (2016) AtVPS41-mediated endocytic pathway is essential for pollen tube-stigmainteraction in Arabidopsis. PNAS 113: 6307-12.


Harris, R. J., and Spellman, M. W. (1993) O-linked fucose and other post-translation modifications unique to EGF module. Glycobiology3: 219-24.


Hellens, R. P., Edwards, E. A., Leyland, N. R., Bean, S., and Mullineaux, P. M. (2000) pGreen: a versatile and flexible binary Ti vector forAgrobacterium-mediated plant transformation. Plant Mol. Biol. 42: 819-32.


Jiang, L., Yang, S. L., Xie, L. F., Puah, C. S., Zhang, X. Q., Yang, W. C., Sundarsen, V., Ye, D. (2005) VANGUARD1 encodes a pectinmethylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract. Plant Cell 17: 584-96.


Kohorn, B. D. (2016) Cell wall-associated kinases and pectin perception. J. Exp. Bot. 67: 489-94.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Kovall, R. A., Gebelein, B., Sprinzak, D., and Kopan, R. (2017) The canonical Notch signaling pathway: Structure and biochemicalinsights into shape, sugar, and force. Dev. Cell. 41: 228-241.


Leonhard-Melief, C., and Haltiwanger, R. S. (2010) O-fucosylation of thrombospondin type 1 repeats. Methods Enzymol. 480: 401-16.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Levitin, B., Richter, D., Markovich, I., and Zik, M. (2008) Arabinogalactan proteins 6 and 11 are required for stamen and pollen functionin Arabidopsis. Plant J. 56: 351-63.


Leydon, A. R., Tsukamoto, T., Dunatunga, D., Qin, Y., Johnson, M. A., Palanivelu, R. (2015) Pollen tube discharge completes theprocess of synergid degeneration that is initiated by pollen tube-synergid interaction in Arabidopsis. Plant Physiol. 169: 485-96.


Lira-Navarette, E., Valero-Gonzalez, J., Villanueva, R., Martinez-Julvez, M., Tejero, T., Merino, P., Panjikar, S., and Hurtado-Guerrero,R. (2011) Structural insights into the mechanism of protein O-fucosylation. PLoS ONE 6: e25365.


Luca, V. C., Kim, B. C., Ge, C., Kakuda, S., Wu, D., Roein-Peikar, M., et al. (2017) Notch-Jagged complex structure implicates a catchbond in tuning ligand sensitivity. Science 355: 1320-24.



http://www.ncbi.nlm.nih.gov/pubmed?term=Goldman%2C M%2E H%2E S%2E%2C Goldberg%2C R%2E B%2E%2C and Mariani%2C C%2E %281994%29 Female sterile tobacco plants are produced by stigma%2Dspecific cell ablation%2E EMBO J%2E 13%3A 2976%2D84%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Goldman, M. H. S., Goldberg, R. B., and Mariani, C. (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J. 13: 2976-84.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Goldman,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Female sterile tobacco plants are produced by stigma-specific cell ablation&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Female sterile tobacco plants are produced by stigma-specific cell ablation&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Goldman,&as_ylo=1994&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Grefen%2C C%2E%2C Donald%2C N%2E%2C Hashimoto%2C K%2E%2C Kudla%2C J%2E%2C Schumacher%2C K%2E%2C and Blatt%2C M%2E R%2E %282010%29 A ubiquitin%2D10 promoter%2Dbased vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies%2E Plant J%2E 64%3A 355%2D65%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Grefen, C., Donald, N., Hashimoto, K., Kudla, J., Schumacher, K., and Blatt, M. R. (2010) A ubiquitin-10 promoter-based vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies. Plant J. 64: 355-65.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Grefen,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A ubiquitin-10 promoter-based vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A ubiquitin-10 promoter-based vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Grefen,&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Hao%2C L%2E%2C Liu%2C J%2E%2C Zhong%2C S%2E%2C Gu%2C H%2E%2C and Qu%2C L%2E J%2E %282016%29 AtVPS41%2Dmediated endocytic pathway is essential for pollen tube%2Dstigma interaction in Arabidopsis%2E PNAS 113%3A 6307%2D12%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Hao, L., Liu, J., Zhong, S., Gu, H., and Qu, L. J. (2016) AtVPS41-mediated endocytic pathway is essential for pollen tube-stigma interaction in Arabidopsis. PNAS 113: 6307-12.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Hao,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=AtVPS41-mediated endocytic pathway is essential for pollen tube-stigma interaction in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=AtVPS41-mediated endocytic pathway is essential for pollen tube-stigma interaction in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Hao,&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Harris%2C R%2E J%2E%2C and Spellman%2C M%2E W%2E %281993%29 O%2Dlinked fucose and other post%2Dtranslation modifications unique to EGF module%2E Glycobiology 3%3A 219%2D24%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Harris, R. J., and Spellman, M. W. (1993) O-linked fucose and other post-translation modifications unique to EGF module. Glycobiology 3: 219-24.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Harris,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=O-linked fucose and other post-translation modifications unique to EGF module&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=O-linked fucose and other post-translation modifications unique to EGF module&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Harris,&as_ylo=1993&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Hellens%2C R%2E P%2E%2C Edwards%2C E%2E A%2E%2C Leyland%2C N%2E R%2E%2C Bean%2C S%2E%2C and Mullineaux%2C P%2E M%2E %282000%29 pGreen%3A a versatile and flexible binary Ti vector for Agrobacterium%2Dmediated plant transformation%2E Plant Mol%2E Biol%2E 42%3A 819%2D32%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Hellens, R. P., Edwards, E. A., Leyland, N. R., Bean, S., and Mullineaux, P. M. (2000) pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation. Plant Mol. Biol. 42: 819-32.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Hellens,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Hellens,&as_ylo=2000&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Jiang%2C L%2E%2C Yang%2C S%2E L%2E%2C Xie%2C L%2E F%2E%2C Puah%2C C%2E S%2E%2C Zhang%2C X%2E Q%2E%2C Yang%2C W%2E C%2E%2C Sundarsen%2C V%2E%2C Ye%2C D%2E %282005%29 VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract%2E Plant Cell 17%3A 584%2D96%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Jiang, L., Yang, S. L., Xie, L. F., Puah, C. S., Zhang, X. Q., Yang, W. C., Sundarsen, V., Ye, D. (2005) VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract. Plant Cell 17: 584-96.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Jiang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Jiang,&as_ylo=2005&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Kohorn%2C B%2E D%2E %282016%29 Cell wall%2Dassociated kinases and pectin perception%2E J%2E Exp%2E Bot%2E 67%3A 489%2D94%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Kohorn, B. D. (2016) Cell wall-associated kinases and pectin perception. J. Exp. Bot. 67: 489-94.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kohorn,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Cell wall-associated kinases and pectin perception&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Cell wall-associated kinases and pectin perception&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kohorn,&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Kovall%2C R%2E A%2E%2C Gebelein%2C B%2E%2C Sprinzak%2C D%2E%2C and Kopan%2C R%2E %282017%29 The canonical Notch signaling pathway%3A Structure and biochemical insights into shape%2C sugar%2C and force%2E Dev%2E Cell%2E 41%3A 228%2D241%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Kovall, R. A., Gebelein, B., Sprinzak, D., and Kopan, R. (2017) The canonical Notch signaling pathway: Structure and biochemical insights into shape, sugar, and force. Dev. Cell. 41: 228-241.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kovall,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The canonical Notch signaling pathway: Structure and biochemical insights into shape, sugar, and force&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The canonical Notch signaling pathway: Structure and biochemical insights into shape, sugar, and force&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kovall,&as_ylo=2017&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Leonhard%2DMelief%2C C%2E%2C and Haltiwanger%2C R%2E S%2E %282010%29 O%2Dfucosylation of thrombospondin type 1 repeats%2E Methods Enzymol%2E 480%3A 401%2D16%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Leonhard-Melief, C., and Haltiwanger, R. S. (2010) O-fucosylation of thrombospondin type 1 repeats. Methods Enzymol. 480: 401-16.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Leonhard-Melief,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=O-fucosylation of thrombospondin type 1 repeats&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=O-fucosylation of thrombospondin type 1 repeats&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Leonhard-Melief,&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Levitin%2C B%2E%2C Richter%2C D%2E%2C Markovich%2C I%2E%2C and Zik%2C M%2E %282008%29 Arabinogalactan proteins 6 and 11 are required for stamen and pollen function in Arabidopsis%2E Plant J%2E 56%3A 351%2D63%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Levitin, B., Richter, D., Markovich, I., and Zik, M. (2008) Arabinogalactan proteins 6 and 11 are required for stamen and pollen function in Arabidopsis. Plant J. 56: 351-63.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Levitin,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Arabinogalactan proteins 6 and 11 are required for stamen and pollen function in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Arabinogalactan proteins 6 and 11 are required for stamen and pollen function in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Levitin,&as_ylo=2008&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Leydon%2C A%2E R%2E%2C Tsukamoto%2C T%2E%2C Dunatunga%2C D%2E%2C Qin%2C Y%2E%2C Johnson%2C M%2E A%2E%2C Palanivelu%2C R%2E %282015%29 Pollen tube discharge completes the process of synergid degeneration that is initiated by pollen tube%2Dsynergid interaction in Arabidopsis%2E Plant Physiol%2E 169%3A 485%2D96%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Leydon, A. R., Tsukamoto, T., Dunatunga, D., Qin, Y., Johnson, M. A., Palanivelu, R. (2015) Pollen tube discharge completes the process of synergid degeneration that is initiated by pollen tube-synergid interaction in Arabidopsis. Plant Physiol. 169: 485-96.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Leydon,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Pollen tube discharge completes the process of synergid degeneration that is initiated by pollen tube-synergid interaction in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Pollen tube discharge completes the process of synergid degeneration that is initiated by pollen tube-synergid interaction in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Leydon,&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Lira%2DNavarette%2C E%2E%2C Valero%2DGonzalez%2C J%2E%2C Villanueva%2C R%2E%2C Martinez%2DJulvez%2C M%2E%2C Tejero%2C T%2E%2C Merino%2C P%2E%2C Panjikar%2C S%2E%2C and Hurtado%2DGuerrero%2C R%2E %282011%29 Structural insights into the mechanism of protein O%2Dfucosylation%2E PLoS ONE 6%3A e25365%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lira-Navarette, E., Valero-Gonzalez, J., Villanueva, R., Martinez-Julvez, M., Tejero, T., Merino, P., Panjikar, S., and Hurtado-Guerrero, R. (2011) Structural insights into the mechanism of protein O-fucosylation. PLoS ONE 6: e25365.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lira-Navarette,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Structural insights into the mechanism of protein O-fucosylation&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Structural insights into the mechanism of protein O-fucosylation&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lira-Navarette,&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Luca%2C V%2E C%2E%2C Kim%2C B%2E C%2E%2C Ge%2C C%2E%2C Kakuda%2C S%2E%2C Wu%2C D%2E%2C Roein%2DPeikar%2C M%2E%2C et al%2E %282017%29 Notch%2DJagged complex structure implicates a catch bond in tuning ligand sensitivity%2E Science 355%3A 1320%2D24%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Luca, V. C., Kim, B. C., Ge, C., Kakuda, S., Wu, D., Roein-Peikar, M., et al. (2017) Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity. Science 355: 1320-24.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Luca,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Luca,&as_ylo=2017&as_allsubj=all&hl=en&c2coff=1


Luo, Y., and Haltiwanger, R. S. (2005) O-fucosylation of notch occurs in the endoplasmic reticulum. J. Biol. Chem. 280: 11289-94.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Mizukami, A. G., Inatsugi, R., Jiao, J., Kotake, T., Kuwata, K., Ootani, K., et al. (2016) The AMOR arabinogalactan sugar chain inducespollen-tube competency to respond to ovular guidance. Curr. Biol. 26: 1091-7.


Mori T., Kuroiwa H., Higasiyama T., and Kuroiwa T. (2006). Generative Cell Specific 1 is essential for angiosperm fertilization. NatureCell Biology 8: 64-71.


Myers, C., Romanowsky, S. M., Barron, Y. D., Garg, S., Azuse, C. L., Curran, A., Davis, R. M., Hatton, J., Harmon, A. C., and Harper, J. F.(2009) Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes. Plant J. 59: 528-39


Nakagawa, T., Suzuki, T., Murata, S., Nakamura, S., Hino, T., Maeo, K., Tabata, R., Kawai, T., Tanaka, K., Niwa, Y., Watanabe, Y.,Nakamura, K., Kimura, T., and Ishiguro, S. (2007) Improved gateway binary vectors: high-performance vectors for creation of fusionconstructs in transgenic analysis of plants. Biosci. Biotechnol. Biochem. 71: 2095-100.


Neumetzler, L., Humphrey, T., Lumba, S., Snyder, S., Yeats, T. H., Usadel, B., Vasilevski, A., Patel, J., Rose, J. K., Persson, S., andBonetta, D. (2012) The FRIABLE1 gene product affects cell adhesion in Arabidopsis. PLoS ONE 7: e42914.


Okajima, T., Xu, A., and Irvine, K. D. (2003) Modulation of notch-ligand binding by protein O-fucosyltransferase 1 and fringe. J. Biol.Chem. 278: 42340-5.


Okuda, S., Tsutsui, H., Shiina, K., Sprunck, S., Takeuchi, H., Yui, R., et al. (2009) Defensin-like polypeptide LUREs are pollen tubeattractants secreted from the synergid cells. Nature 458: 357-61.


Palanivelu, R., and Johnson, M. A. (2010) Functional genomics of pollen tube-pistil interactions in Arabidopsis. Biochem. Soc. Trans.38: 593-7.


Palanivelu, R., Brass, L., Edlund, A. F., Preuss, D. (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis genethat controls GABA levels. Cell 114: 47-59.


Qin, Y., Leydon, A. R., Manziello, A., Pandey, R., Mount, D., Denic, S., Vasic, B., Johnson, M. A., and Palanivelu, R. (2009) Penetration ofthe stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil. PLoS Genet. 5:1000621.


Rampal, R., Li, A. S., Moloney, D. J., Georgiou, S. A., Luther, K. B., Nita-Lazar, A., and Haltiwanger, R. S. (2005) Lunatic fringe, maniacfringe, and radical fringe recognize similar specificity determinants in O-fucosylated epidermal growth factor-like repeats. J. Biol.Chem. 280: 42454-63.


Rebay, I., Fleming, R. J., Fehon, R. G., Cherbas, L., Cherbas, P., and Artavanis-Tsakonas, S. (1991) Specific EGF repeats of Notchhttps://plantphysiol.orgDownloaded on February 4, 2021. - Published by Copyright (c) 2020 American Society of Plant Biologists. All rights reserved.

http://www.ncbi.nlm.nih.gov/pubmed?term=Luo%2C Y%2E%2C and Haltiwanger%2C R%2E S%2E %282005%29 O%2Dfucosylation of notch occurs in the endoplasmic reticulum%2E J%2E Biol%2E Chem%2E 280%3A 11289%2D94%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Luo, Y., and Haltiwanger, R. S. (2005) O-fucosylation of notch occurs in the endoplasmic reticulum. J. Biol. Chem. 280: 11289-94.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Luo,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=O-fucosylation of notch occurs in the endoplasmic reticulum&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=O-fucosylation of notch occurs in the endoplasmic reticulum&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Luo,&as_ylo=2005&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Mizukami%2C A%2E G%2E%2C Inatsugi%2C R%2E%2C Jiao%2C J%2E%2C Kotake%2C T%2E%2C Kuwata%2C K%2E%2C Ootani%2C K%2E%2C et al%2E %282016%29 The AMOR arabinogalactan sugar chain induces pollen%2Dtube competency to respond to ovular guidance%2E Curr%2E Biol%2E 26%3A 1091%2D7%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Mizukami, A. G., Inatsugi, R., Jiao, J., Kotake, T., Kuwata, K., Ootani, K., et al. (2016) The AMOR arabinogalactan sugar chain induces pollen-tube competency to respond to ovular guidance. Curr. Biol. 26: 1091-7.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Mizukami,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The AMOR arabinogalactan sugar chain induces pollen-tube competency to respond to ovular guidance&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The AMOR arabinogalactan sugar chain induces pollen-tube competency to respond to ovular guidance&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Mizukami,&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Mori T%2E%2C Kuroiwa H%2E%2C Higasiyama T%2E%2C and Kuroiwa T%2E %282006%29%2E Generative Cell Specific 1 is essential for angiosperm fertilization%2E Nature Cell Biology 8%3A 64%2D71%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Mori T., Kuroiwa H., Higasiyama T., and Kuroiwa T. (2006). Generative Cell Specific 1 is essential for angiosperm fertilization. Nature Cell Biology 8: 64-71.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Mori&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Generative Cell Specific 1 is essential for angiosperm fertilization&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Generative Cell Specific 1 is essential for angiosperm fertilization&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Mori&as_ylo=2006&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Myers%2C C%2E%2C Romanowsky%2C S%2E M%2E%2C Barron%2C Y%2E D%2E%2C Garg%2C S%2E%2C Azuse%2C C%2E L%2E%2C Curran%2C A%2E%2C Davis%2C R%2E M%2E%2C Hatton%2C J%2E%2C Harmon%2C A%2E C%2E%2C and Harper%2C J%2E F%2E %282009%29 Calcium%2Ddependent protein kinases regulate polarized tip growth in pollen tubes%2E Plant J%2E 59%3A 528%2D39&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Myers, C., Romanowsky, S. M., Barron, Y. D., Garg, S., Azuse, C. L., Curran, A., Davis, R. M., Hatton, J., Harmon, A. C., and Harper, J. F. (2009) Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes. Plant J. 59: 528-39

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Myers,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Myers,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Nakagawa%2C T%2E%2C Suzuki%2C T%2E%2C Murata%2C S%2E%2C Nakamura%2C S%2E%2C Hino%2C T%2E%2C Maeo%2C K%2E%2C Tabata%2C R%2E%2C Kawai%2C T%2E%2C Tanaka%2C K%2E%2C Niwa%2C Y%2E%2C Watanabe%2C Y%2E%2C Nakamura%2C K%2E%2C Kimura%2C T%2E%2C and Ishiguro%2C S%2E %282007%29 Improved gateway binary vectors%3A high%2Dperformance vectors for creation of fusion constructs in transgenic analysis of plants%2E Biosci%2E Biotechnol%2E Biochem%2E 71%3A 2095%2D100%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Nakagawa, T., Suzuki, T., Murata, S., Nakamura, S., Hino, T., Maeo, K., Tabata, R., Kawai, T., Tanaka, K., Niwa, Y., Watanabe, Y., Nakamura, K., Kimura, T., and Ishiguro, S. (2007) Improved gateway binary vectors: high-performance vectors for creation of fusion constructs in transgenic analysis of plants. Biosci. Biotechnol. Biochem. 71: 2095-100.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Nakagawa,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Improved gateway binary vectors: high-performance vectors for creation of fusion constructs in transgenic analysis of plants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Improved gateway binary vectors: high-performance vectors for creation of fusion constructs in transgenic analysis of plants&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Nakagawa,&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Neumetzler%2C L%2E%2C Humphrey%2C T%2E%2C Lumba%2C S%2E%2C Snyder%2C S%2E%2C Yeats%2C T%2E H%2E%2C Usadel%2C B%2E%2C Vasilevski%2C A%2E%2C Patel%2C J%2E%2C Rose%2C J%2E K%2E%2C Persson%2C S%2E%2C and Bonetta%2C D%2E %282012%29 The FRIABLE1 gene product affects cell adhesion in Arabidopsis%2E PLoS ONE 7%3A e42914%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Neumetzler, L., Humphrey, T., Lumba, S., Snyder, S., Yeats, T. H., Usadel, B., Vasilevski, A., Patel, J., Rose, J. K., Persson, S., and Bonetta, D. (2012) The FRIABLE1 gene product affects cell adhesion in Arabidopsis. PLoS ONE 7: e42914.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Neumetzler,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The FRIABLE1 gene product affects cell adhesion in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The FRIABLE1 gene product affects cell adhesion in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Neumetzler,&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Okajima%2C T%2E%2C Xu%2C A%2E%2C and Irvine%2C K%2E D%2E %282003%29 Modulation of notch%2Dligand binding by protein O%2Dfucosyltransferase 1 and fringe%2E J%2E Biol%2E Chem%2E 278%3A 42340%2D5%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Okajima, T., Xu, A., and Irvine, K. D. (2003) Modulation of notch-ligand binding by protein O-fucosyltransferase 1 and fringe. J. Biol. Chem. 278: 42340-5.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Okajima,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Modulation of notch-ligand binding by protein O-fucosyltransferase 1 and fringe&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Modulation of notch-ligand binding by protein O-fucosyltransferase 1 and fringe&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Okajima,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Okuda%2C S%2E%2C Tsutsui%2C H%2E%2C Shiina%2C K%2E%2C Sprunck%2C S%2E%2C Takeuchi%2C H%2E%2C Yui%2C R%2E%2C et al%2E %282009%29 Defensin%2Dlike polypeptide LUREs are pollen tube attractants secreted from the synergid cells%2E Nature 458%3A 357%2D61%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Okuda, S., Tsutsui, H., Shiina, K., Sprunck, S., Takeuchi, H., Yui, R., et al. (2009) Defensin-like polypeptide LUREs are pollen tube attractants secreted from the synergid cells. Nature 458: 357-61.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Okuda,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Defensin-like polypeptide LUREs are pollen tube attractants secreted from the synergid cells&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Defensin-like polypeptide LUREs are pollen tube attractants secreted from the synergid cells&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Okuda,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Palanivelu%2C R%2E%2C and Johnson%2C M%2E A%2E %282010%29 Functional genomics of pollen tube%2Dpistil interactions in Arabidopsis%2E Biochem%2E Soc%2E Trans%2E 38%3A 593%2D7%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Palanivelu, R., and Johnson, M. A. (2010) Functional genomics of pollen tube-pistil interactions in Arabidopsis. Biochem. Soc. Trans. 38: 593-7.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Palanivelu,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Functional genomics of pollen tube-pistil interactions in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Functional genomics of pollen tube-pistil interactions in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Palanivelu,&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Palanivelu%2C R%2E%2C Brass%2C L%2E%2C Edlund%2C A%2E F%2E%2C Preuss%2C D%2E %282003%29 Pollen tube growth and guidance is regulated by POP2%2C an Arabidopsis gene that controls GABA levels%2E Cell 114%3A 47%2D59%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Palanivelu, R., Brass, L., Edlund, A. F., Preuss, D. (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 114: 47-59.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Palanivelu,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Palanivelu,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Qin%2C Y%2E%2C Leydon%2C A%2E R%2E%2C Manziello%2C A%2E%2C Pandey%2C R%2E%2C Mount%2C D%2E%2C Denic%2C S%2E%2C Vasic%2C B%2E%2C Johnson%2C M%2E A%2E%2C and Palanivelu%2C R%2E %282009%29 Penetration of the stigma and style elicits a novel transcriptome in pollen tubes%2C pointing to genes critical for growth in a pistil%2E PLoS Genet%2E 5%3A 1000621%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Qin, Y., Leydon, A. R., Manziello, A., Pandey, R., Mount, D., Denic, S., Vasic, B., Johnson, M. A., and Palanivelu, R. (2009) Penetration of the stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil. PLoS Genet. 5: 1000621.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Qin,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Penetration of the stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Penetration of the stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Qin,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Rampal%2C R%2E%2C Li%2C A%2E S%2E%2C Moloney%2C D%2E J%2E%2C Georgiou%2C S%2E A%2E%2C Luther%2C K%2E B%2E%2C Nita%2DLazar%2C A%2E%2C and Haltiwanger%2C R%2E S%2E %282005%29 Lunatic fringe%2C maniac fringe%2C and radical fringe recognize similar specificity determinants in O%2Dfucosylated epidermal growth factor%2Dlike repeats%2E J%2E Biol%2E Chem%2E 280%3A 42454%2D63%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Rampal, R., Li, A. S., Moloney, D. J., Georgiou, S. A., Luther, K. B., Nita-Lazar, A., and Haltiwanger, R. S. (2005) Lunatic fringe, maniac fringe, and radical fringe recognize similar specificity determinants in O-fucosylated epidermal growth factor-like repeats. J. Biol. Chem. 280: 42454-63.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Rampal,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Lunatic fringe, maniac fringe, and radical fringe recognize similar specificity determinants in O-fucosylated epidermal growth factor-like repeats&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Lunatic fringe, maniac fringe, and radical fringe recognize similar specificity determinants in O-fucosylated epidermal growth factor-like repeats&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Rampal,&as_ylo=2005&as_allsubj=all&hl=en&c2coff=1


mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor. Cell 67: 687-99.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Sandaklie-Nikolova, L., Palanivelu, R., King, E. J., Copenhavaer, G. P., and Drews, G. N. (2007) Synergid cell death in Arabidopsis istriggered following direct interaction with the pollen tube. Plant Physiol. 144: 1753-62.


Sasamura, T., Sasaki, N., Miyashita, F., Nakao, S., Ishikawa, H. O., Ito, M., Kitagawa, M., Harigaya, K., Spana, E., Bilder, D., Perrimon, N.,and Matsuno, K. (2003) Neurotic, a novel maternal neurogenic gene, encodes an O-fucosyltransferase that is essential for Notch-Deltainteractions. Development 130: 4785-95.


Schiott, M., Romanowsky, S. M., Baekgaard, L., Jakobsen, M. K., Palmgren, M. G., and Harper, J. F. (2004) A plant plasma membranecalcium pump is required for normal pollen tube growth and fertilization. PNAS 101: 9502-7.


Sessions, R. A., Zambryski, P. C. (1995) Arabidopsis gynoecium structure in the wild and in ettin mutants. Development 121: 1519-32.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Shi, S., and Stanley, P. (2003) Protein O-fucosyltransferase 1 is an essential component of Notch signaling pathways. PNAS 100: 5234-9.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Stahl, M., Uemura, K., Ge, C., Shi, S., Tashima, Y., and Stanley, P. (2008) Roles of Pofut1 and O-fucose in mammalian Notch signaling. J.Biol. Chem. 283: 13638-51.


Takeuchi, H., Higashiyama, T. (2012) A species-specific cluster of defensing-like genes encodes diffusible pollen tube attractants inArabidopsis. PLoS Biol. 10: e1001449.


Takeuchi, H., Higashiyama, T. (2016) Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis. Nature 531:245-8.


Tanz, S. K., Castleden, I., Hooper, C. M., Vacher, M., Small, I., and Millar, H. A. (2013) SUBA3: a database for integrating experimentationand prediction to define the SUBcellular location of proteins in Arabidopsis. Nucleic Acids Res. 41: D1185-91.


Trindade, H., Boavida, L. C., Borralho, N., and Fiejo, J. A. (2001) Successful fertilization and seed set from pollination on immature non-dehisced flowers of Eucalyptus globulus. Ann. Bot. 87: 469-475.


van Tuyl, J. M., van Dien, M. P., van Creij, M. G. M., van Kleinwee, T. C. M., Franken, J., and Bino, R. J. (1991) Application of in vitropollination, ovary cultures, ovule culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses. PlantSci. 74: 115-26.


Verger, S., Chabout, S., Gineau, E., and Mouille, G. (2016) Cell adhesion in plant is under the control of putative O-fucosyltransferases.Development 143: 2536-40.

Pubmed: Author and TitleCrossRef: Author and Title https://plantphysiol.orgDownloaded on February 4, 2021. - Published by


http://www.ncbi.nlm.nih.gov/pubmed?term=Rebay%2C I%2E%2C Fleming%2C R%2E J%2E%2C Fehon%2C R%2E G%2E%2C Cherbas%2C L%2E%2C Cherbas%2C P%2E%2C and Artavanis%2DTsakonas%2C S%2E %281991%29 Specific EGF repeats of Notch mediate interactions with Delta and Serrate%3A implications for Notch as a multifunctional receptor%2E Cell 67%3A 687%2D99%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Rebay, I., Fleming, R. J., Fehon, R. G., Cherbas, L., Cherbas, P., and Artavanis-Tsakonas, S. (1991) Specific EGF repeats of Notch mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor. Cell 67: 687-99.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Rebay,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Specific EGF repeats of Notch mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Specific EGF repeats of Notch mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Rebay,&as_ylo=1991&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Sandaklie%2DNikolova%2C L%2E%2C Palanivelu%2C R%2E%2C King%2C E%2E J%2E%2C Copenhavaer%2C G%2E P%2E%2C and Drews%2C G%2E N%2E %282007%29 Synergid cell death in Arabidopsis is triggered following direct interaction with the pollen tube%2E Plant Physiol%2E 144%3A 1753%2D62%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Sandaklie-Nikolova, L., Palanivelu, R., King, E. J., Copenhavaer, G. P., and Drews, G. N. (2007) Synergid cell death in Arabidopsis is triggered following direct interaction with the pollen tube. Plant Physiol. 144: 1753-62.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sandaklie-Nikolova,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Synergid cell death in Arabidopsis is triggered following direct interaction with the pollen tube&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Synergid cell death in Arabidopsis is triggered following direct interaction with the pollen tube&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sandaklie-Nikolova,&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Sasamura%2C T%2E%2C Sasaki%2C N%2E%2C Miyashita%2C F%2E%2C Nakao%2C S%2E%2C Ishikawa%2C H%2E O%2E%2C Ito%2C M%2E%2C Kitagawa%2C M%2E%2C Harigaya%2C K%2E%2C Spana%2C E%2E%2C Bilder%2C D%2E%2C Perrimon%2C N%2E%2C and Matsuno%2C K%2E %282003%29 Neurotic%2C a novel maternal neurogenic gene%2C encodes an O%2Dfucosyltransferase that is essential for Notch%2DDelta interactions%2E Development 130%3A 4785%2D95%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Sasamura, T., Sasaki, N., Miyashita, F., Nakao, S., Ishikawa, H. O., Ito, M., Kitagawa, M., Harigaya, K., Spana, E., Bilder, D., Perrimon, N., and Matsuno, K. (2003) Neurotic, a novel maternal neurogenic gene, encodes an O-fucosyltransferase that is essential for Notch-Delta interactions. Development 130: 4785-95.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sasamura,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Neurotic, a novel maternal neurogenic gene, encodes an O-fucosyltransferase that is essential for Notch-Delta interactions&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Neurotic, a novel maternal neurogenic gene, encodes an O-fucosyltransferase that is essential for Notch-Delta interactions&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sasamura,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Schiott%2C M%2E%2C Romanowsky%2C S%2E M%2E%2C Baekgaard%2C L%2E%2C Jakobsen%2C M%2E K%2E%2C Palmgren%2C M%2E G%2E%2C and Harper%2C J%2E F%2E %282004%29 A plant plasma membrane calcium pump is required for normal pollen tube growth and fertilization%2E PNAS 101%3A 9502%2D7%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Schiott, M., Romanowsky, S. M., Baekgaard, L., Jakobsen, M. K., Palmgren, M. G., and Harper, J. F. (2004) A plant plasma membrane calcium pump is required for normal pollen tube growth and fertilization. PNAS 101: 9502-7.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Schiott,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A plant plasma membrane calcium pump is required for normal pollen tube growth and fertilization&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A plant plasma membrane calcium pump is required for normal pollen tube growth and fertilization&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Schiott,&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Sessions%2C R%2E A%2E%2C Zambryski%2C P%2E C%2E %281995%29 Arabidopsis gynoecium structure in the wild and in ettin mutants%2E Development 121%3A 1519%2D32%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Sessions, R. A., Zambryski, P. C. (1995) Arabidopsis gynoecium structure in the wild and in ettin mutants. Development 121: 1519-32.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sessions,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Arabidopsis gynoecium structure in the wild and in ettin mutants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Arabidopsis gynoecium structure in the wild and in ettin mutants&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sessions,&as_ylo=1995&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Shi%2C S%2E%2C and Stanley%2C P%2E %282003%29 Protein O%2Dfucosyltransferase 1 is an essential component of Notch signaling pathways%2E PNAS 100%3A 5234%2D9%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Shi, S., and Stanley, P. (2003) Protein O-fucosyltransferase 1 is an essential component of Notch signaling pathways. PNAS 100: 5234-9.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Shi,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Protein O-fucosyltransferase 1 is an essential component of Notch signaling pathways&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Protein O-fucosyltransferase 1 is an essential component of Notch signaling pathways&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Shi,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Stahl%2C M%2E%2C Uemura%2C K%2E%2C Ge%2C C%2E%2C Shi%2C S%2E%2C Tashima%2C Y%2E%2C and Stanley%2C P%2E %282008%29 Roles of Pofut1 and O%2Dfucose in mammalian Notch signaling%2E J%2E Biol%2E Chem%2E 283%3A 13638%2D51%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Stahl, M., Uemura, K., Ge, C., Shi, S., Tashima, Y., and Stanley, P. (2008) Roles of Pofut1 and O-fucose in mammalian Notch signaling. J. Biol. Chem. 283: 13638-51.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Stahl,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Roles of Pofut1 and O-fucose in mammalian Notch signaling&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Roles of Pofut1 and O-fucose in mammalian Notch signaling&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Stahl,&as_ylo=2008&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Takeuchi%2C H%2E%2C Higashiyama%2C T%2E %282012%29 A species%2Dspecific cluster of defensing%2Dlike genes encodes diffusible pollen tube attractants in Arabidopsis%2E PLoS Biol%2E 10%3A e1001449%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Takeuchi, H., Higashiyama, T. (2012) A species-specific cluster of defensing-like genes encodes diffusible pollen tube attractants in Arabidopsis. PLoS Biol. 10: e1001449.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Takeuchi,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A species-specific cluster of defensing-like genes encodes diffusible pollen tube attractants in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A species-specific cluster of defensing-like genes encodes diffusible pollen tube attractants in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Takeuchi,&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Takeuchi%2C H%2E%2C Higashiyama%2C T%2E %282016%29 Tip%2Dlocalized receptors control pollen tube growth and LURE sensing in Arabidopsis%2E Nature 531%3A 245%2D8%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Takeuchi, H., Higashiyama, T. (2016) Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis. Nature 531: 245-8.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Takeuchi,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Takeuchi,&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Tanz%2C S%2E K%2E%2C Castleden%2C I%2E%2C Hooper%2C C%2E M%2E%2C Vacher%2C M%2E%2C Small%2C I%2E%2C and Millar%2C H%2E A%2E %282013%29 SUBA3%3A a database for integrating experimentation and prediction to define the SUBcellular location of proteins in Arabidopsis%2E Nucleic Acids Res%2E 41%3A D1185%2D91%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Tanz, S. K., Castleden, I., Hooper, C. M., Vacher, M., Small, I., and Millar, H. A. (2013) SUBA3: a database for integrating experimentation and prediction to define the SUBcellular location of proteins in Arabidopsis. Nucleic Acids Res. 41: D1185-91.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Tanz,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Tanz,&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Trindade%2C H%2E%2C Boavida%2C L%2E C%2E%2C Borralho%2C N%2E%2C and Fiejo%2C J%2E A%2E %282001%29 Successful fertilization and seed set from pollination on immature non%2Ddehisced flowers of Eucalyptus globulus%2E Ann%2E Bot%2E 87%3A 469%2D475%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Trindade, H., Boavida, L. C., Borralho, N., and Fiejo, J. A. (2001) Successful fertilization and seed set from pollination on immature non-dehisced flowers of Eucalyptus globulus. Ann. Bot. 87: 469-475.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Trindade,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Successful fertilization and seed set from pollination on immature non-dehisced flowers of Eucalyptus globulus&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Successful fertilization and seed set from pollination on immature non-dehisced flowers of Eucalyptus globulus&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Trindade,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=van Tuyl%2C J%2E M%2E%2C van Dien%2C M%2E P%2E%2C van Creij%2C M%2E G%2E M%2E%2C van Kleinwee%2C T%2E C%2E M%2E%2C Franken%2C J%2E%2C and Bino%2C R%2E J%2E %281991%29 Application of in vitro pollination%2C ovary cultures%2C ovule culture%2C and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses%2E Plant Sci%2E 74%3A 115%2D26%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=van Tuyl, J. M., van Dien, M. P., van Creij, M. G. M., van Kleinwee, T. C. M., Franken, J., and Bino, R. J. (1991) Application of in vitro pollination, ovary cultures, ovule culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses. Plant Sci. 74: 115-26.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=van&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Application of in vitro pollination, ovary cultures, ovule culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Application of in vitro pollination, ovary cultures, ovule culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=van&as_ylo=1991&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Verger%2C S%2E%2C Chabout%2C S%2E%2C Gineau%2C E%2E%2C and Mouille%2C G%2E %282016%29 Cell adhesion in plant is under the control of putative O%2Dfucosyltransferases%2E Development 143%3A 2536%2D40%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Verger, S., Chabout, S., Gineau, E., and Mouille, G. (2016) Cell adhesion in plant is under the control of putative O-fucosyltransferases. Development 143: 2536-40.


Google Scholar: Author Only Title Only Author and Title

Villalobos, J. A., Yi, B. R., and Wallace, I. S. (2015) 2-fluoro-L-fucose is a metabolically incorporated inhibitor of plant cell wallpolysaccharide biosynthesis. PLoS ONE 10: e0139091


Wang, Y., Shao, L., Shi, S., Harris, R. J., Spellman, M. W., Stanley, P., and Haltiwanger, R. S. (2001) Modification of epidermal growthfactor-like repeats with O-fucose: Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase. J. Biol.Chem. 276: 40338-45.


Wang, Y., Mortimer, J. C., Davis, J., Dupree, P., and Keegstra, K. (2013) Identification of an additional protein involved in mannanbiosynthesis. Plant J. 73: 105-17.


Wilson, Z. A., Morroll, S. M., Dawson, J., Swarup, R., and Tighe, P. J. (2001) The Arabidopsis MALE STERILITY (MS1) gene is atranscriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J. 28: 27-39.


Zentella, R., Sui, N., Barnhill, B., Hsieh, W. P., Hu, J., Shabanowitz, J., et al. (2017) The Arabidopsis O-fucosyltransferase SPINDLYactivates nuclear growth repressor DELLA. Nat. Chem. Biol. 13: 479-85.



http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Verger,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Cell adhesion in plant is under the control of putative O-fucosyltransferases&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Cell adhesion in plant is under the control of putative O-fucosyltransferases&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Verger,&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Villalobos%2C J%2E A%2E%2C Yi%2C B%2E R%2E%2C and Wallace%2C I%2E S%2E %282015%29 2%2Dfluoro%2DL%2Dfucose is a metabolically incorporated inhibitor of plant cell wall polysaccharide biosynthesis%2E PLoS ONE 10%3A e0139091&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Villalobos, J. A., Yi, B. R., and Wallace, I. S. (2015) 2-fluoro-L-fucose is a metabolically incorporated inhibitor of plant cell wall polysaccharide biosynthesis. PLoS ONE 10: e0139091

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Villalobos,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=2-fluoro-L-fucose is a metabolically incorporated inhibitor of plant cell wall polysaccharide biosynthesis.&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=2-fluoro-L-fucose is a metabolically incorporated inhibitor of plant cell wall polysaccharide biosynthesis.&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Villalobos,&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Wang%2C Y%2E%2C Shao%2C L%2E%2C Shi%2C S%2E%2C Harris%2C R%2E J%2E%2C Spellman%2C M%2E W%2E%2C Stanley%2C P%2E%2C and Haltiwanger%2C R%2E S%2E %282001%29 Modification of epidermal growth factor%2Dlike repeats with O%2Dfucose%3A Molecular cloning and expression of a novel GDP%2Dfucose protein O%2Dfucosyltransferase%2E J%2E Biol%2E Chem%2E 276%3A 40338%2D45%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Wang, Y., Shao, L., Shi, S., Harris, R. J., Spellman, M. W., Stanley, P., and Haltiwanger, R. S. (2001) Modification of epidermal growth factor-like repeats with O-fucose: Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase. J. Biol. Chem. 276: 40338-45.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Wang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Modification of epidermal growth factor-like repeats with O-fucose: Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Modification of epidermal growth factor-like repeats with O-fucose: Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Wang%2C Y%2E%2C Mortimer%2C J%2E C%2E%2C Davis%2C J%2E%2C Dupree%2C P%2E%2C and Keegstra%2C K%2E %282013%29 Identification of an additional protein involved in mannan biosynthesis%2E Plant J%2E 73%3A 105%2D17%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Wang, Y., Mortimer, J. C., Davis, J., Dupree, P., and Keegstra, K. (2013) Identification of an additional protein involved in mannan biosynthesis. Plant J. 73: 105-17.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Wang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Identification of an additional protein involved in mannan biosynthesis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Identification of an additional protein involved in mannan biosynthesis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang,&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Wilson%2C Z%2E A%2E%2C Morroll%2C S%2E M%2E%2C Dawson%2C J%2E%2C Swarup%2C R%2E%2C and Tighe%2C P%2E J%2E %282001%29 The Arabidopsis MALE STERILITY %28MS1%29 gene is a transcriptional regulator of male gametogenesis%2C with homology to the PHD%2Dfinger family of transcription factors%2E Plant J%2E 28%3A 27%2D39%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Wilson, Z. A., Morroll, S. M., Dawson, J., Swarup, R., and Tighe, P. J. (2001) The Arabidopsis MALE STERILITY (MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J. 28: 27-39.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Wilson,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis MALE STERILITY (MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis MALE STERILITY (MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wilson,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=Zentella%2C R%2E%2C Sui%2C N%2E%2C Barnhill%2C B%2E%2C Hsieh%2C W%2E P%2E%2C Hu%2C J%2E%2C Shabanowitz%2C J%2E%2C et al%2E %282017%29 The Arabidopsis O%2Dfucosyltransferase SPINDLY activates nuclear growth repressor DELLA%2E Nat%2E Chem%2E Biol%2E 13%3A 479%2D85%2E&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Zentella, R., Sui, N., Barnhill, B., Hsieh, W. P., Hu, J., Shabanowitz, J., et al. (2017) The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA. Nat. Chem. Biol. 13: 479-85.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zentella,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zentella,&as_ylo=2017&as_allsubj=all&hl=en&c2coff=1


a putative protein o-fucosyltransferase facilitates pollen ...feb 21, 2018 · 20 one sentence...

Documents