vesicular trafficking and cell expansion during maize leaf development

1
Program/Abstract # 484 Global analysis of the LEAFY transcriptional network: Transitioning to reproductive development in Arabidopsis thaliana Cara Winter, Doris Wagner Department of Biology, University of Pennsylvania, USA The transition from vegetative to reproductive development in Arabidopsis thaliana involves the integration of multiple environ- mental and endogenous cues and the translation of these cues into the developmental program of ower-morphogenesis. The LEAFY (LFY) transcription factor has been identied as the key integrator and master regulator of this transition. In addition to its role in the ini- tiation of oral development during the meristem identity (MI) transition, LFY also has a later role in the induction of homeotic genes that specify oral organ identity (i.e. oral patterning). Using a post- translationally activatable form of LFY, a ChIP-on-chip approach was taken to identify targets of LFY during the initiation of ower deve- lopment. Preliminary analyses suggest that LFY binds to over 1000 targets, suggesting it plays a large role in the transition to reproductive development in Arabidopsis. All known MI regulators were identied as LFY targets in this dataset. In addition, new potential LFY targets include genes involved in leaf development, hormone signaling, chromatin remodeling, and owering time, suggesting a link between meristem identity and these pathways. doi:10.1016/j.ydbio.2008.05.465 Program/Abstract # 485 The histone acetyltransferase GCN5 affects oral bud initiation and stamen development in Arabidopsis Amy T. Hark, Ross Cohen, John Schocken, Elizabeth R. McCain Biology Department, Muhlenberg College, Allentown, PA 18104, USA A central question in biology is to understand how gene expression is precisely regulated to give rise to a variety of forms during the process of development. Epigenetic effects such as DNA methylation or histone modication have been increasingly shown to play a critical role in regulation of genome function. GCN5 is prototypical histone acetyltransferase that has been shown to participate in regulating developmental gene expression in a several metazoan species. In Ar- abidopsis, plants with T-DNA insertions in GCN5 (also known as HAG1) display a variety of pleiotropic effects including dwarsm, loss of apical dominance, and oral defects affecting fertility. We sought to determine when during early development these oral defects rst arise. Using scanning electron microscopy, we demonstrate that gcn5- 1/hag 1-1 and gcn5-4/hag1-5 mutants display abnormal proliferation of young buds around the oral meristem as well as defects in stamen number and arrangement at later stages. This provides temporal and spatial information to aid in the identication of GCN5 targets in the developing ower. doi:10.1016/j.ydbio.2008.05.466 Program/Abstract # 486 Morphogenesis, meristems and maize: Genetic regulation of inorescence development in plants Paula McSteen, Andrea Skirpan, Solmaz Barazesh, Xianting Wu, Kim Phillips Department of Biology, Penn State University, University Park, PA, USA Organogenesis in plants is controlled by meristems. Meristems are organized groups of cells analogous to animal stem cellsthat divide and differentiate to give rise to all of the organs of the plant. In particular, axillary or lateral meristems give rise to branches and owers and therefore play a fundamental role in plant morphogenesis and reproduction. My lab focuses on the initiation of axillary meri- stems in the inorescence (owering branches) of the genetic model organism, maize. We have shown that the plant growth hormone, auxin, which plays a critical role in cell division, cell expansion and polar growth, is required for axillary meristem initiation during maize inorescence development. We have identied and characterized a large class of mutants, called barren inorescence, which are defective in the initiation of axillary meristems. Cloning of these genes by our lab and others has discovered genes required for auxin biosynthesis, transport and response. Analysis of the genetic, molecular, cell biology and biochemical interactions between these genes has led to the elucidation of a genetic network regulating axillary meristem initiation in the maize inorescence. This research was funded by NSF IOB-0416616 and USDA 2007-03036. doi:10.1016/j.ydbio.2008.05.467 Program/Abstract # 487 Vesicular trafcking and cell expansion during maize leaf development Daniel Hill a , Xingyuan Ling a , Anding Luo a , Mike Tamkun b , Anne Sylvester a a Department of Molecular Biology, University Wyoming, Laramie, WY, USA b Department of Biomedical Sciences, CSU, Fort Collins, CO, USA Maize leaves grow in a predictable and repetitive pattern from the shoot apical meristem. Cell shapes in maize leaves directly reect the history of division orientation relative to directional cell expansion during early development and it has been shown that the balance of division and expansion contributes to the nal leaf shape. We are interested in understanding how these growth processes are regu- lated and have taken a forward genetics approach to identify mutants with altered cell shapes and sizes due to lesions in genes that regulate division and/or expansion. A screen identied the Warty1 (Wty1) locus, which results in abnormal cell growth when it is mutated. Transposon tagging showed that this locus encodes ZmRAB2A1, a highly conserved, small GTPase. Homologous Rab2 proteins across diverse eukaryotes are known to function in vesicular trafcking within a restricted ER and Golgi cellular compartment. Using the re- cently released maize genome sequence, we classied 47 maize RABS into 8 distinct sub-groups. The ZmRAB2 sub-group bears high se- quence similarity to HsRab2, although the maize sub-group has more members. Phenotypic characteristics of the wty1-0 mutant, and unique aspects of grass cell expansion, suggest that ZmRAB2A1 may have acquired monocot-specic functions. This hypothesis is sup- ported by live-cell imaging of YFP-tagged ZmRAB2A1 trafcking in multiple compartments, including the cortical cytoplasm. This novel localization suggests diversied function of ZmRAB2A1 and implicates the vesicle trafcking pathway mediated by ZmRAB2A1 in cellular signaling during leaf expansion. doi:10.1016/j.ydbio.2008.05.468 610 ABSTRACTS / Developmental Biology 319 (2008) 599610

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Program/Abstract # 484Global analysis of the LEAFY transcriptional network: Transitioningto reproductive development in Arabidopsis thalianaCara Winter, Doris WagnerDepartment of Biology, University of Pennsylvania, USA

The transition from vegetative to reproductive development inArabidopsis thaliana involves the integration of multiple environ-mental and endogenous cues and the translation of these cues into thedevelopmental program of flower-morphogenesis. The LEAFY (LFY)transcription factor has been identified as the key integrator andmaster regulator of this transition. In addition to its role in the ini-tiation of floral development during the meristem identity (MI)transition, LFY also has a later role in the induction of homeotic genesthat specify floral organ identity (i.e. floral patterning). Using a post-translationally activatable form of LFY, a ChIP-on-chip approach wastaken to identify targets of LFY during the initiation of flower deve-lopment. Preliminary analyses suggest that LFY binds to over 1000targets, suggesting it plays a large role in the transition to reproductivedevelopment in Arabidopsis. All known MI regulators were identifiedas LFY targets in this dataset. In addition, new potential LFY targetsinclude genes involved in leaf development, hormone signaling,chromatin remodeling, and flowering time, suggesting a link betweenmeristem identity and these pathways.

doi:10.1016/j.ydbio.2008.05.465

Program/Abstract # 485The histone acetyltransferase GCN5 affects floral bud initiationand stamen development in ArabidopsisAmy T. Hark, Ross Cohen, John Schocken, Elizabeth R. McCainBiology Department, Muhlenberg College, Allentown, PA 18104, USA

A central question in biology is to understand how gene expressionis precisely regulated to give rise to a variety of forms during theprocess of development. Epigenetic effects such as DNA methylationor histonemodification have been increasingly shown to play a criticalrole in regulation of genome function. GCN5 is prototypical histoneacetyltransferase that has been shown to participate in regulatingdevelopmental gene expression in a several metazoan species. In Ar-abidopsis, plants with T-DNA insertions in GCN5 (also known as HAG1)display a variety of pleiotropic effects including dwarfism, loss ofapical dominance, and floral defects affecting fertility. We sought todetermine when during early development these floral defects firstarise. Using scanning electron microscopy, we demonstrate that gcn5-1/hag 1-1 and gcn5-4/hag1-5 mutants display abnormal proliferationof young buds around the floral meristem as well as defects in stamennumber and arrangement at later stages. This provides temporal andspatial information to aid in the identification of GCN5 targets in thedeveloping flower.

doi:10.1016/j.ydbio.2008.05.466

Program/Abstract # 486Morphogenesis, meristems and maize: Genetic regulation ofinflorescence development in plantsPaula McSteen, Andrea Skirpan, Solmaz Barazesh, Xianting Wu,Kim PhillipsDepartment of Biology, Penn State University, University Park, PA, USA

Organogenesis in plants is controlled by meristems. Meristems areorganized groups of cells analogous to animal “stem cells” that divideand differentiate to give rise to all of the organs of the plant. Inparticular, axillary or lateral meristems give rise to branches andflowers and therefore play a fundamental role in plant morphogenesisand reproduction. My lab focuses on the initiation of axillary meri-stems in the inflorescence (flowering branches) of the genetic modelorganism, maize. We have shown that the plant growth hormone,auxin, which plays a critical role in cell division, cell expansion andpolar growth, is required for axillary meristem initiation during maizeinflorescence development. We have identified and characterized alarge class of mutants, called “barren inflorescence”, which aredefective in the initiation of axillary meristems. Cloning of thesegenes by our lab and others has discovered genes required for auxinbiosynthesis, transport and response. Analysis of the genetic,molecular, cell biology and biochemical interactions between thesegenes has led to the elucidation of a genetic network regulatingaxillary meristem initiation in the maize inflorescence. This researchwas funded by NSF IOB-0416616 and USDA 2007-03036.

doi:10.1016/j.ydbio.2008.05.467

Program/Abstract # 487Vesicular trafficking and cell expansion during maize leafdevelopmentDaniel Hill a, Xingyuan Ling a, Anding Luo a, Mike Tamkun b, AnneSylvester a

a Department of Molecular Biology, University Wyoming, Laramie, WY,USAb Department of Biomedical Sciences, CSU, Fort Collins, CO, USA

Maize leaves grow in a predictable and repetitive pattern from theshoot apical meristem. Cell shapes in maize leaves directly reflect thehistory of division orientation relative to directional cell expansionduring early development and it has been shown that the balance ofdivision and expansion contributes to the final leaf shape. We areinterested in understanding how these growth processes are regu-lated and have taken a forward genetics approach to identify mutantswith altered cell shapes and sizes due to lesions in genes that regulatedivision and/or expansion. A screen identified the Warty1 (Wty1)locus, which results in abnormal cell growth when it is mutated.Transposon tagging showed that this locus encodes ZmRAB2A1, ahighly conserved, small GTPase. Homologous Rab2 proteins acrossdiverse eukaryotes are known to function in vesicular traffickingwithin a restricted ER and Golgi cellular compartment. Using the re-cently released maize genome sequence, we classified 47 maize RABSinto 8 distinct sub-groups. The ZmRAB2 sub-group bears high se-quence similarity to HsRab2, although the maize sub-group has moremembers. Phenotypic characteristics of the wty1-0 mutant, andunique aspects of grass cell expansion, suggest that ZmRAB2A1 mayhave acquired monocot-specific functions. This hypothesis is sup-ported by live-cell imaging of YFP-tagged ZmRAB2A1 trafficking inmultiple compartments, including the cortical cytoplasm. This novellocalization suggests diversified function of ZmRAB2A1 and implicatesthe vesicle trafficking pathway mediated by ZmRAB2A1 in cellularsignaling during leaf expansion.

doi:10.1016/j.ydbio.2008.05.468

610 ABSTRACTS / Developmental Biology 319 (2008) 599–610