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Pierce's Disease FDA Regulatory Framework - Gabriel Paulino - Pierce's Disease Symposium 2008
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Grape: Vitis vinifera
Entering the Regulatory Coordinated Framework
Outline
• PIPRA and CDFA PD/GWSS Board• Grape Industry• Grape Biology• The Problem: Pierce’s Disease• The Solution: Strategies with High Potential
CDFA PD/GWSS Board• CDFA PD/GWSS Board created in 2001 after the
Temecula Valley disaster• Funded from California growers and state and
federal governments money• Fund basic research programs to understand the
fundamental biology of the pathogen: Xylella fastidiosa and the vector: Glassy-Winged Sharpshooter
• Goal develop PD control strategies for the grape industry
PIPRA
• PIPRA = Public Intellectual Property Resource for Agriculture• supports agricultural innovation for both
humanitarian and specialty crop commercial purposes
• bring together intellectual property from over 53 universities, public agencies, and non-profit institutes
www.pipra .org
PIPRA and CDFA PD/GWSS Board
• Since 2005, PIPRA has supported the CDFA PD/GWSS Board in:– Intellectual property management– Policy development for IP management– Research Management Tool (
www.piercesdisease.org)– More recently, Regulatory Assistance for Field
Trial Launch and GM-based strategies for PD control
Categories Total
Wine, Grape and Grape Products contribution to US Economy
$162 Billion
Employment 1.1 Million equivalent full-time employment
Agriculture
Grape Growers 23,486
Grape Bearing Acres 934,750
Farm Gate Grape Sales $3.5 Billion (Highest Value Crop in US/ Largest Fruit Industry in CA)
Taxes $17.1 Billion (9.1 Federal + 8 State and Local)
GRAPE + CITRUS + NURSERIES Industries Revenues
$20.8 Billion (Second after Corn: $26.8 and before Soybean: $18.3)
Categories TotalWine Industry
Direct ImpactWineries 4,929 (2005) in all 50 States
Wineries Sales Revenue $11.4 BillionAdded Value
Distribution Share of American Wine Revenue
$2.7 Billion
Retail and Restaurant Share of American Wine Revenue
$9.8 Billion
Wine-Related Tourism $27.3 MillionWine-Related Tourism Expenditure $3.0 Billion
Grape Production
Product share for grapes in the United States (2000-2006 avg.). Source: USDA Economic Research Service
Grape Distribution• Perennial woody vines in the genus
Vitis, family Vitaceae• According to the USDA data
(http:/plants.usda.gov) 23 Vitis species exist in the United States and Canada
• The most common wild species found in the United States are Summer Grapes, the Frost Grape, the Sweet Winter Grape, the Northern Fox Grape, the Catbird Grape and the Riverbank Grape.
Grape Biology
• Cultivated varieties= propagated by grafting shoots (scions) onto the root of a hardy plant (rootstocks)
• Desirable traits=displayed by the scion not the rootstock
• Self-fruitful => pollination by wind and lesser extent insects
• Fruit = berries; small (<1 inch)http://www.extension.umn.edu/yardandgarden/YGLNews/images/scion&rootstock.jpg
Grape, Papaya & Plum Consumption
• Wine consumption is 2.47 gallons per year (2007)
Total Fresh Processed
Total (lbs)
Fresh Total Canned Juice Dried
Grape 21.14 8.02 13.12 0.14 6.00 6.97Papaya 1.08 1.08 N/APlum 2.03 1.01 1.02 0.02 0.55 0.45
Source: USDA/Economic Research Service. Data last updated February 27, 2009.
Grape Food/Feed Use
Pierce’s Disease : The threat
• Was first discovered by Newton Barris Pierce in 1892 in Anaheim
• Was not a big threat in California until the introduction of the Glassy Winged-Sharpshooter in 1994
• $37.9 million loss in Riverside and San Diego counties in 1998/99 due to vine death and removal of vineyards => triggered the creation of the CDFA PD/GWSS Board which was completely industry-driven in 2000
Glassy-Winged Sharpshooter: The vector
• Originated from Southeast US and Northeast Mexico
• First reported in 1994 in Southern California
• Feeds on xylem fluid of a large number of plants
• Aggressive flyer, traveling greater distances than native sharpshooters
Why is the GWSS a bigger threat?
• Wide host range• Fly further inside the grape fields• Ability to feed on and transmit the pathogen
to older grape wood.
Important Vectors of Xf
Glassy-winged sharpshooterHomalodisca coagulata
Blue-green sharpshooter Graphocephala atropunctata Redheaded sharpshooter
Carneocephala fulgida
Green SharpshooterDraeculacephala minerva
Pierce’s Disease
Gram negative rod fastidious
Insect vectored
Xylem limited
PD symptoms similar to waterstress
Pierce’s Disease
Oleander Leaf Scorch Coffee Leaf Scorch
Almond Leaf Scorch
Maple Leaf ScorchMaple Leaf Scorch
Elm Leaf ScorchOak Leaf Scorch
Phony Peach FruitNo leaf scorching
CVC fruit and stunted growthNo leaf Scorching
Leaf scorch symptom
Pierce’s Disease Symptoms
Berry desiccation
“Green Islands” Irregular periderm development
“Matchstick” petioles
Pierce’s Disease Symptoms
Chronically infected vines have delayed growth and stunted shoots
Varela et al. 2001
Specialty Crop PD Threat on Crop Production Value ($)
Grape 4.0 Billion
Almond 2.8 Billion
Citrus 1.1 Billion
Stone Fruit 1.0 Billion
PD Threatens Other Crop Industries
Summary
• Grape Industry is an important for the US Agricultural Economy
• The Pierce’s Disease is threatening the California Grape/Wine Industry since 2000
• The CDFA PD/GWSS Board has invested $270 Million of research funding to find a solution
• Major research findings are finally giving some hope to the industry
Strategies with high potential of commercialization
2 Main strategies
• Barrier Crop: Use the GM Crop as a barrier crop border to protect grapevine crop
• GM Grape Rootstocks: Develop GM-Grape rootstock with PD-Resistance => Non-GM Grape scions can still be grafted with their own wine and grape qualities
2 Development Strategies
• 1TDNA Plant Transformation with Gene of Interest and Plant Selection Gene:– Gene of Interest conferring PD resistance from:
baculovirus, bacteria, grape and heterologous plants
– Selectable Markers: kanamycin, hygromycin and visual marker GUS
PIPRA’s recombinase-based marker-free transformation platform
• Recombinase-based marker excision system => GM Vitis marker free
Figure 1. Negative selection construct to test efficacy Par-A mediated recombinase activity in tobacco and grape.
Genomic DNA
Selectable Marker Cassette
RB
Recombinase Cassette
RRS
RRS
Gene of Interest Cassette: GUS
LB
DsRed fluorescece protein Marker Cassette
A. Recombinase-based Marker Excision Module
B. Recombinase-mediated Excision
Genomic DNA
RB
RRS
Gene of Interest Cassette: GUS
LB
Selectable Marker Cassette
Recombinase Cassette
RRS
RRS
DsRed fluorescece protein Marker Cassette
Lost DNA
Diffusible Signaling Factor (DSF)• DSF = small fatty acid that accumulates when Xf
is high density in xylem
• Strategy = altering DSF levels in a grape rootstock to decrease Xf virulence by a process called pathogen confusion
• Gene of Interest = rpfF/B from Xylella or Xanthomonas
Poly-Galacturonase Inhibiting Protein (PGIP)
• Xf Polygalacturonase = Cell Wall Degrading Enzyme allowing movement and dispersion of Xf between xylem vessels in the plant
• Strategy = Expressing PG-Inhibiting Proteins directly in a grape rootstock to inhibit Xf PGs
• Gene of Interest = PGIP from heterologous plant
Anti-Programmed Cell Death (PCD)
• Symptoms = Activation of Programmed Cell Death Pathways (PCD)
• Strategy = Suppression of PD Symptoms by constitutively expressing anti-PCD genes
• Gene of Interest = anti-PCD genes from bacoluvirus, bacteria, grape or heterologous plants
Comparison of constructs
Discussion• Short-term objectives:
– First field trials planned for Spring 2010• What information needed for field permit and interstate
movement?
– Regulatory Road-Map• Evaluate the different GM strategies in terms of Cost/Time
– TDNA – Recombinase-based Marker-free System– GM Rootstock– GM Grapevines as Border Barrier
Discussion
• Long-term Objectives:– Data requirements for deregulation of a TDNA vs.
Marker-free developed strategy?
– Data requirements for deregulation of a GM Grapevine vs. GM Rootstock?