Development of cdna microarrays for gene expression research in floriculture crops
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Development of cDNA Microarrays for Gene Expression Research in Floriculture Crops. Dave Clark University of Florida Environmental Horticulture Dept. UF Floriculture Biotech Lab (est. 1995). Our main goal: To bridge the gap between basic molecular biology and applied floriculture

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Development of cdna microarrays for gene expression research in floriculture crops

Development of cDNA Microarrays for Gene Expression Research in Floriculture Crops

Dave Clark

University of Florida

Environmental Horticulture Dept.


Uf floriculture biotech lab est 1995
UF Floriculture Biotech Lab in Floriculture Crops(est. 1995)

  • Our main goal:

  • To bridge the gap between basic molecular biology and applied floriculture

    • Florida produces 20% of US floriculture crops


Uf floriculture biotech
UF Floriculture Biotech in Floriculture Crops

Development:

  • Phase 1 – 1995-2000 (then)

    • Biotechnology

  • Phase 2 – 2000-2005 (now)

    • Functional Genomics

  • Phase 3 – 2005-2010 (next)

    • Gene Function


Phase 1 1995 2000
Phase 1 – 1995-2000 in Floriculture Crops

  • Establish a biotech system

    • Petunia hybrida

  • Proof of concept research

    • Expansion of available technology

  • Build critical personnel mass

    • Recruit good people

    • Make strong collaborations


Phase 1 1995 20001
Phase 1 – 1995-2000 in Floriculture Crops

  • Transgenic Plants - hormones

  • Gibberellins

    • Dwarf plants

  • Cytokinins

    • “Stay Green” leaves

  • Ethylene

    • Long lasting flowers


Internal growth regulators
Internal Growth Regulators in Floriculture Crops

AtGA2ox7

Makes dwarf tobacco

Schomburg et al. 2003


Internal growth regulators1
Internal Growth Regulators in Floriculture Crops

AtGA2ox7 - Makes dwarf petunia


Stay green leaves
Stay Green Leaves in Floriculture Crops

sag-IPT

Makes leaves stay green longer in tobacco

Gan & Amasino 1995


Sag ipt
sag-IPT in Floriculture Crops

Makes leaves stay greener longer in petunia

Clark et al. in press


Sag ipt drought tolerance
sag-IPT in Floriculture Crops – Drought Tolerance


Long Lasting Flowers in Floriculture Crops

Etr1-1

ETR1-1 - Makes plants ethylene insensitive


Long Lasting Flowers in Floriculture Crops

Control

Etr1-1


Problems with 35s etr1 1
Problems with 35S:: in Floriculture Cropsetr1-1

  • etr1-1 petunias – commercially limited

rooting

pathogens – etr1-1

‘MD’

etr1-1


Conclusions phase 1
Conclusions – Phase 1 in Floriculture Crops

  • We have a good experimental system

  • We have good people in the lab

  • We have good collaborators

  • We needdifferent genes

  • We needbetter promoters


Phase 2 2000 2005
Phase 2 – 2000-2005 in Floriculture Crops

  • Functional Genomics

    • New tools for genetics research

  • Exploration for new technologies

    • New and novel genes

  • Controlling gene expression

    • Petal specific promoters


Functional genomics
Functional Genomics in Floriculture Crops

  • Tools that help discover the biological function of genes

  • Uncovers how groups of genes work together in a biological process

  • Encompasses many traditional biological and genetic approaches


Petunia EST Project in Floriculture Crops

(EST = Expressed Sequence Tags)

mRNA extraction

Tissue collection

Bioinformatics and Microarrays

library synthesis

EST sequencing


Petunia flower genes
Petunia Flower Genes in Floriculture Crops

  • Petunia gene sequencing:

    Floral development library 2774

    C2H4-treated flower library 3264

    Pollinated flower library 1824

    Other libraries 1046

  • Total genes sequenced 8908


Petunia est database
Petunia EST Database in Floriculture Crops


Microarray Construction in Floriculture Crops

4200 unique petunia genes


Microarray analysis – Ethylene Regulated Genes in Floriculture Crops

Green A>B

Yellow A=B

Red A<B

Control

(A)

Ethylene

(B)

A+B


Ethylene regulated genes
Ethylene Regulated Genes in Floriculture Crops

  • We have isolated approximately 50 genes that are regulated by ethylene

  • Some of these genes are thought to be involved in making floral scent volatiles

    • Benzoic acid carboxyl methyltransferase


Phase 2 still going
Phase 2 – Still going in Floriculture Crops

  • We still bridge the gap between basic molecular biology and applied floriculture

    • We have a petunia EST database

    • Microarrays are working well

    • We have many new and novel genes

    • We have good promoters


Phase 3 2005 2010
Phase 3 – 2005-2010 in Floriculture Crops

  • Gene Function

    • We are ahead of schedule:

    • Experimental system (phase 1)

      • Adopted by several groups

    • New genetic tools (phase 2)

      • Available to floriculture researchers

    • Understanding gene function


Measuring floral scent
Measuring Floral Scent in Floriculture Crops


Flower scent composition petunia
Flower Scent Composition - Petunia in Floriculture Crops

Benzoic acid carboxyl methyltransferase (BAMT) - makes methyl benzoate in petunia


Stop and smell the roses
Stop and Smell the Roses! in Floriculture Crops

US consumers bought almost 130 million roses on Valentine’s Day 2003

US consumers spent over $7.5B on cut flowers in 2002


Opportunity knocks
Opportunity Knocks! in Floriculture Crops

  • Commercial cut rose varieties don’t have much fragrance


Flower Scent Composition - Rose in Floriculture Crops


The future
The Future in Floriculture Crops

  • We plan to maintain and grow the genetics database

  • We plan to make transgenic plants expressing new genes

  • We plan to knock out 1 gene a week – functional analysis

  • We plan to continue making new collaborations


Who supports us
Who supports us? in Floriculture Crops

  • Intellectually

    • Harry Klee – UF

    • Bill Farmerie – UF

    • Don McCarty – UF

    • Michelle Jones – OSU

    • Jim Giovannoni – USDA/Cornell

    • EU PetNet Group

  • Financially

    • USDA Floriculture Initiative

    • American Floral Endowment

    • Fred Gloeckner Foundation

    • Florida FIRST

    • The Scotts Company


Uf floriculture biotechnology 2002 2003
UF Floriculture Biotechnology in Floriculture Crops2002-2003


Questions? in Floriculture Crops


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