tam66274: ultra-low gossypol cottonseed keerti s. rathore · 2019-08-02 · tam66274: ultra-low...
TRANSCRIPT
TAM66274: Ultra-low Gossypol Cottonseed
Keerti S. Rathore
Institute for Plant Genomics and Biotechnology
& Dept. of Soil & Crop Sciences
Texas A&M University System
Cotton Breeders Tour, Texas A&M Univ.
July 23, 2019
Production Exports
World 25,970
China 6,532 16
India 6,432 849
USA 3,553 2,330
Pakistan 2,308 120
Burkina Faso 290 256 (Gold: #1 and Cotton: #2)
Mali 231 152 (Gold: #1 and Cotton: #2)
Cote d'Ivoire 183 174
Cameroon 109 98
Benin 131 125 (Cotton: #1)
Chad 48 41
Togo 38 33
Tanzania 82 44
Egypt 114 33
Zimbabwe 54 38
Nigeria 44 11
Cotton Production and Exports 1,000 Metric Tons (2014-15)
Sources: FAS/USDA - May, 2015; PRB – 2016; http://atlas.media.mit.edu/en/
Production Global Hunger Index
World 25,970
China 6,532 25th
India 6,432 103rd
Pakistan 2,308 106th
Burkina Faso 290 89th
Mali 231 90th
Cote d'Ivoire 183 85th
Cameroon 109 71st
Benin 131 80th
Chad 48 118th
Togo 38 80th
Tanzania 82 95th
Egypt 114 61st
Zimbabwe 54 107th
Nigeria 44 103rd
Cotton Production 1,000 Metric Tons (2014-15) &
Global Hunger Index (2018)
Sources: FAS/USDA - May, 2015; PRB – 2016; http://atlas.media.mit.edu/en/
Rampant protein-calorie malnutrition in many poor countries
- cereal-based diets do not provide enough protein
• Brown rice 7.3% Protein (75 - 95% calories in some countries)
• Maize 9.8%
• Millet 11.5%
• Sorghum 8.3%
• Wheat 10.6%
• Cassava 1.0%
• Yam 2.0%
Ultra-low Gossypol Cottonseed (ULGCS)
From Conception to Deregulation of TAM66274
(https://www.aphis.usda.gov/brs/aphisdocs/17_29201p.pdf)
Global Cotton Lint Production in 2014 = 26 MMT
Global Cottonseed Production in 2014 = 47 MMT (~10.5 MMT Protein)
Can meet the protein requirements of 575 million people/year (50 g/day)
Presence of toxic gossypol prevents the use of cottonseed as food &
also limits its use as feed
• Protein content of the seed is about 22.5%; good quality protein.
• Its use for human consumption or animal feed is limited by the presence of gossypol in the seed glands.
• Gossypol is toxic to non-ruminant animals, including pigs, chicken and humans. It damages heart and liver.
• Meal used as feed for the cattle – gossypol binds to lysine thus reducing the nutritional value of the meal.
• Gossypol typically removed during the processing of cottonseed to obtain edible oil.
Gossypol
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
• Cottonseed Oil (gossypol removed during the processing)
• Feed for cattle either directly as seed or as meal
following oil extraction (in limited quantities)
Current Use of Cottonseed in Human Nutrition
Gossypol is Toxic to Monogastric Animals
• 3.66 kg of crude protein is converted to 0.83 kg of
milk protein per day (PCR = 4.4)
• Feed conversion ratio (FCR): 5.8
• Protein Conversion Ratio (PCR): 20
Cows are Inefficient in Converting Crude Protein into Animal Protein
FCR: Feed consumed / weight gain
PCR: Feed Protein consumed/ Edible Animal Protein Produced
Protein Conversion Ratio
20
5.7 4.7
8.85.74.6
Tilman and Clark (2014) Nature 515:518
Claude Boyd (2005) Global Aquaculture Advocate 73-74
2.6
7.7
• McMichael discovered a GLANDLESS mutant of cotton
in 1954
• Seed from glandless cotton was gossypol-free
• National and International breeding programs launched to transfer the trait to commercial varieties
• Animal nutrition studies showed glandless cottonseed to be a relatively good source of feed for pigs, chicken, and shrimp
• Glandless cottonseed was even considered fit for
human consumption
Attempts to Eliminate Gossypol Using Plant Breeding
Wild-type ‘Glanded’ Cotton Mutant ‘Glandless’ Cotton
Gossypol: 10,000 ppm Gossypol: 0-10 ppm
Gossypol and other related terpenoid aldehydes present in the glands in leaves, stem, and floral parts confer resistance to insect pests
These terpenoids are also induced in response to pathogens and serve as phytoalexins
Traditional Insect Pests that Cause Greater Damage to
Glandless Cotton
Cotton Bollworm (Helicoverpa zea)
Tobacco Budworm (Heliothis virescens)
Beet Armyworm (Spodoptera exigua)
Non-traditional Insects that Damage Glandless Cotton
Black Fleahopper (Spanogonicus albofasciatus)
Pillbug (Porcellia sp.)
Spotted Cucumber Beetle (Diabrotica undecimpunctata)
Grape Colaspis (Maecolaspis flavida)
Green Dock Beetle (Gastrophysa cyanea)
Blister Beetle (Epicauta vittata)
▪ Currently, there is no large-scale cultivation of glandless cotton.
Desired Trait in Cotton
Seeds:Gossypol <450 ppm (0.45 μg/mg)
(FDA Guidelines)
Leaf & other Organs:
Full Compliment of Gossypol
and Related Terpenoids
for Defense Against
Insects and Diseases
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
XO
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
XO
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
XO
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
X8-Hydroxy-(+)-
(HGQ)
dCS
(+)--Cadinene-Cadinene
Hemigossypolone
(dHG)Desoxyhemigossypol
(HG)Hemigossypol
(G)
Gossypol
-OcimeneMyrcene
Farnesyldiphosphate
+ +
(FDP)
Heliocide H2 Heliocide H3 Heliocide H1 Heliocide H4
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
XO
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
dCH
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
dCS = -cadinene synthase dCH = -cadinene hydroxylase
(G)
Gossypol
O
HO
HO
HO
HO
CHO
OH
CHO
HO
HO
O
O
CHO
HO
HO
O
O
+
Desoxyhemigossypol
(dHG)
Heliocide H2
Heliocide H3
Hemigossypol
(HG)
+
CHO
HO
HO
O
O
Hemigossypolone
(HGQ)
CHO
HO
HO
O
O
Heliocide H1
CHO
HO
HO
O
O
HeliocideH4
Myrcene
-Ocimene
HO
HO
CHO
OH
OH
OH
HCO
OH
Gossypol
(G)
OH
8-Hydroxy-(+)-
-cadinene
(+)--CadineneOPP
Farnesyldiphosphate
(FDP)
dCS
X
(Cytosol)Mevalonate (MVA) pathway
IPP
DMAPP
Seed-specific, RNAi-mediated Silencing of δ-cadinene synthase Gene
(α-Globulin Promoter::RNAi against δ-Cadinene Synthase)
0
50
100
150
200
250
300
350
400
Stem Root Pollen SeedLeaf Floral
bud
GU
S A
CT
IVIT
Y
(nm
ole
4-M
U /
mg p
rote
in/
min
)
Seed-specific expression of AGP::gusAin cotton
0
50
100
150
200
250
300
350
400
Control
AGP:gusA
Sunilkumar …… Rathore (2002)
Transgenic Research 11: 347-359
U.S. Patent #7,626,081
0
2
4
6
8
10
12
Wild-type
3 4 5 6 7 8 9 10
14
LCT66-2
mg
go
ss
yp
ol
mg
-1ti
ss
ue
LCT66-32
10.45 0.54 0.54 0.07* 0.46 0.13*
(n=10) (n=8) (n=8)
1 2 3 4 5 6 7 8 9 101 2 3 4 5 6 7 8 9 101 2
Gossypol Levels in Individual Wild-type
and T1 Cottonseeds
Sunilkumar …… Rathore (2006)
PNAS 103:18054-18059
U.S. Patent #7999148
Wild-type Null LCT66-2-4 LCT66-32-7
9.65 0.459.19 0.19(n=10) (n=10)
0
2
4
6
8
10
12
14
mg
go
ss
yp
ol
mg
-1ti
ss
ue
0.48 0.02*(n=50)
0.61 0.001*
(n=50)
Gossypol Levels in Individual Wild-type,
Null Segregant, and T2 Cottonseeds
Sunilkumar …… Rathore (2006)
PNAS 103:18054-18059
U.S. Patent #7999148
6.466.35
7.48
0.15 0.14 0.140.31 0.21 0.38
0.45
0
1
2
3
4
5
6
7
8
2009 2010 2011
ug
goss
ypo
l/m
g se
ed
-ke
rne
l tis
sue
***
Coker 312 66-49B 66-81 66-250
Coker 312
66-49B
66-81
66-250
ULGCS Trait is stable under field conditionsPalle ----- Rathore (2013) Plant Biotech. J.
Seed gossypol content of field-grown plants
mg
te
rpe
no
ids
mg
-1ti
ss
ue
1.4
1.2
1.0
0.8
0.6
0.4
0.2
0.0
Bract
0.72
0.60
0.48
0.36
0.24
0.12
0.00
Floral Bud
1.8
1.5
1.2
0.9
0.6
0.3
0.0
Boll
mg
te
rpe
no
ids
mg
-1ti
ss
ue
G HGQ H
Terminal bud
Root
5
4
3
2
1
0
G
Petal
8
6
4
2
0
G MG DMG
2.5
2.0
1.5
1.0
0.5
0.0
G HGQ H G HGQ H
G HGQ H dHG dMHG
0.10
0.08
0.06
0.04
0.02
0.00HG MHG
0.6
0.5
0.4
0.3
0.2
0.1
0.0
Levels of Gossypol and Related Terpenoids in
Terminal Bud, Floral Bud, Bract, Petal, Boll, and
Root Tissues
Wild-type Control
LCT66-2
LCT66-32
ULGCS trait is completely tissue specificSunilkumar …… Rathore (2006)
PNAS 103:18054-18059
U.S. Patent #7999148
Deregulation of TAM66274
(Steps since 2006)
✓ Generate many more additional events, test for desired
phenotype
✓ Molecular analyses – Southerns, insert localization,
silencing of the target gene
✓ Biochemical analysis
✓ Inheritance analysis
✓ Agronomic performance trials (8 trials in 3 states)
✓ Seed composition analysis
✓ Fiber quality analysis
✓ USDA-APHIS petition & FDA dossier submitted (2017)
Regular Cotton TAM66274
TAM66274: Ultra-low Gossypol Cottonseed (ULGCS)
(Seed-specific, RNAi silencing of δ-cadinene synthase gene)
• Gossypol levels reduced from ~10,000 ppm to ~300 ppm, well below levels
considered safe for human consumption by FDA (450 ppm) and
FAO/WHO (600 ppm)
• Foliage and Floral parts contain normal levels of gossypol for protection
against pests
https://www.aphis.usda.gov/brs/aphisdocs/17_29201p.pdf
➢ Deregulated by USDA-APHIS
(Oct. 17, 2018)
➢ Awaiting FDA approval
Commercial CottonSeed Gossypol: 10,000 ppm
Insect Protected Foliage
Glandless CottonSeed Gossypol: 10 ppm
No insect protection
TAM66274Seed Gossypol: 300 ppm
Insect Protected Foliage & Floral Tissue
Seed k
ern
el
Leaf
Problem: Seed
Gossypol
Plant Breeding Genetic Engineering
Can gene editing technology (CRISPR/Cas9)
be used to achieve the same objective?
▪ Targeting the δ-cadinene synthase gene will not work
(large gene family, RNA-seq analysis shows expression
of at least 10 genes in the embryo)
▪ Even if another, single copy target gene is identified,
simple knockout of this gene will not achieve
tissue specificity
▪ CRISPR interference is a possibility, but the plant will
still be considered transgenic because of the need for
tissue-specific expression of guide-RNA and CRISPR
through a transgenic cassette
STV474 x TAM66274 – BC3F4 Planted for
Seed Increase in North Carolina in 2019
▪ Feed for poultry and swine
▪ Feed for aquaculture species
▪ Direct use as food
(fortify flour for protein enrichment,
Paruthi Paal)
5.74.7
5.7
4.6
8.8
Possible uses of meal from ULGCS to combat protein malnutrition
2.6
7.7
Rising Demand for Feed by Poultry Industry (Broiler Production)
FAOStat (2019)
Rising Demand for Feed by Poultry Industry (Egg Production)
Protein in Global Cottonseed Output = Protein in 1.7 Trillion Eggs
Global Annual Egg Production = 1.4 Trillion FAOStat (2019)
Source: The State of Fisheries and Aquaculture; FAO 2018
Aquaculture Meets Fish Demand as Ocean Catches Decline
▪ Aquaculture is the fastest growing food production system globally (8% growth)
▪ Aquaculture projected to supply >60% of fish for human consumption by 2030
▪ Demand for fishmeal and fish oil for aquaculture expected to increase,
however, the supplies likely to decrease
➢ Fishmeal price will rise by 90%
➢ Fish oil price will rise by 70%
▪ Great need for substitutes for fishmeal and fish oil
➢ ULGCS meal (41% protein) and oil could serve as a good substitute
➢ Some cottonseed is already used in aquafeeds
Shrimp Feeding Study: Richardson et al. (2016)
Southern Flounder Feeding Study: Alam et al. (2018)
World Bank Report (2013) #83177-GLB
Agriculture and Rural Development Department, The World Bank Group (2011)
Rising Demand for Feed by Aquaculture Industry
➢ Used comparative transcriptomics between normal,
glanded cotton and glandless (Hopi) cotton 14 dpa embryos
to identify three gene pairs involved in the development of
glands in cotton
➢ Cotton Gland Formation (CGF1, CGF2 & CGF3) genes
encode transcription factors
➢ Only the CGF3 genes show sequence differences between
glanded and glandless cotton
Janga et al. Plant Biotech. J. (2019)
Identification and Cloning of Genes Involved in
the Development of Glands in Cotton
Janga et al. Plant Biotech. J. (2019)
Gla
nded
A-G
en
om
e
Gla
ndle
ss
Gla
nded
D-G
en
om
e
Gla
ndle
ss
CGF3, the Main Gene Involved in Gland Development in Cotton
B
Co
ntr
ol
CG
F3
KO
CG
F2
KO
CRISPR-Cas9-mediated, targeted knockout of CGF2 & CGF3 genes
Janga et al. Plant Biotech. J. (2019)
µg
terp
en
oid
/g t
issu
e
L254
D254
Overexpression of CGF3 Gene in Cotton Cultures
HG: Hemigossypol
dHG: Desoxyhemigossypol
HGAL: Hemigossylic acid lactone
MHG: Methoxyhemigossypol
dMHG: Desoxymethoxy-hemigossypol
Goss: Gossypol
MG: Methoxygossypol
DMG: Dimethoxygossypol
New tool to increase glands (gossypol)
in the foliage & flowers to boost the natural
defenses of the cotton plant
Janga et al. Plant Biotech. J. (2019)
USDA-ARS
Robert Stipanovic
Lorraine Puckhaber
Alois Bell
J. Liu
IPGB, TAMU
G. Sunilkumar
LeAnne Campbell
Sabarinath Sundaram
Sylvain Marcel
Sreenath P. Reddy
Devendra Pandeya
Lauren Tollack
Shanna Sherwood
Madhusudhana Janga
Sam Reddy
Robert A. Creelman (John Mullet)
Cotton Inc., TxCOT, TFFC, THECB & Texas AgriLife Research
C. Wayne Smith
Steve Hague
Cotton Inc.
Tom Wedegaertner
Kater Hake
Questions???