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

PIPRA and CDFA PD/GWSS Board

Grape Industry

Grape Biology

The Problem: Pierce’s Disease

The Solution: Strategies with High Potential

Who are the agencies? USDA/APHIS = United States Department of Agriculture / Animal and Plant Health Inspection Service EPA = Environmental Protection Agency FDA = Food and Drug Administration

USDA/APHIS = United States Department of Agriculture / Animal and Plant Health Inspection Service

EPA = Environmental Protection Agency

FDA = Food and Drug Administration

Role of USDA/APHIS Protecting Agriculture from Pest and Disease Regulates the import, handling, interstate movement, and release into the environment of regulated organisms that are products of biotechnology, including organisms undergoing confined experimental use or field trials.

Protecting Agriculture from Pest and Disease

Regulates the import, handling, interstate movement, and release into the environment of regulated organisms that are products of biotechnology, including organisms undergoing confined experimental use or field trials.

Role of EPA regulates the sale, distribution and use of pesticides in order to protect health, and the environment, regardless of how the pesticide was made or its mode of action: Pesticides Biopesticides Plant-Incorporated Protectants (PIPs)

regulates the sale, distribution and use of pesticides in order to protect health, and the environment, regardless of how the pesticide was made or its mode of action:

Pesticides

Biopesticides

Plant-Incorporated Protectants (PIPs)

Role of FDA responsible for ensuring the safety and proper labeling of all plant-derived foods and feeds, including those developed through bioengineering

responsible for ensuring the safety and proper labeling of all plant-derived foods and feeds, including those developed through bioengineering

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

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 = 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

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

Grape Industry Wine, grape and grape products contribute to $162 billion in US Economy (Wine Institute). Employment: 1.1 million full-time equivalent jobs Agriculture: 23,856 grape growers, 934,750 grape bearing acres, $3.5 billion in farm gate grape sales Wine Industry Direct Impact: 4,929 wineries in 2005, up from 2,904 in 2000, a 70% increase in five years; wineries now in all 50 states; $11.4 billion in winery sales revenues

Wine, grape and grape products contribute to $162 billion in US Economy (Wine Institute).

Employment: 1.1 million full-time equivalent jobs

Agriculture: 23,856 grape growers, 934,750 grape bearing acres, $3.5 billion in farm gate grape sales

Wine Industry Direct Impact: 4,929 wineries in 2005, up from 2,904 in 2000, a 70% increase in five years; wineries now in all 50 states; $11.4 billion in winery sales revenues

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 Total Wine Industry Direct Impact Wineries 4,929 (2005) in all 50 States Wineries Sales Revenue $11.4 Billion Added 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 Million Wine-Related Tourism Expenditure $3.0 Billion

Grape Industry Wine Industry Value Added: $2.7 billion in distributor share of American wine revenue; $9.8 billion in retail and restaurant share of American wine revenue; 27.3 million wine-related tourist visits; $3 billion estimated wine-related tourism expenditures. Other Grape Products: $1.669 billion retail value of grape juice and grape product sales; $3 billion retail value of table grape sales; $560 million retail value of raisin sales Total Taxes Paid: $17.1 billion, including $9.1 billion federal and $8 billion state and local

Wine Industry Value Added: $2.7 billion in distributor share of American wine revenue; $9.8 billion in retail and restaurant share of American wine revenue; 27.3 million wine-related tourist visits; $3 billion estimated wine-related tourism expenditures.

Other Grape Products: $1.669 billion retail value of grape juice and grape product sales; $3 billion retail value of table grape sales; $560 million retail value of raisin sales

Total Taxes Paid: $17.1 billion, including $9.1 billion federal and $8 billion state and local

Grape Industry Wine + table + raisin grape revenues = $3 Billion => Highest Value Fruit Crop in US = Largest Fruit Industry in California Grape + Citrus + Nurseries Industries revenues = $20.8 Billion = Second largest revenues of agricultural sector behind corn ($26.8B) and before soybean ($18.3B)

Wine + table + raisin grape revenues

= $3 Billion => Highest Value Fruit Crop in US

= Largest Fruit Industry in California

Grape + Citrus + Nurseries Industries revenues

= $20.8 Billion

= Second largest revenues of agricultural sector behind corn ($26.8B) and before soybean ($18.3B)

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.

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

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)

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 6.97 Papaya 1.08 1.08 N/A Plum 2.03 1.01 1.02 0.02 0.55 0.45 Source: USDA/Economic Research Service. Data last updated February 27, 2009.

Wine consumption is 2.47 gallons per year (2007)

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

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

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.

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 sharpshooter Homalodisca coagulata Blue-green sharpshooter Graphocephala atropunctata Redheaded sharpshooter Carneocephala fulgida Green Sharpshooter Draeculacephala minerva

Pierce’s Disease Gram negative rod fastidious Insect vectored Xylem limited PD symptoms similar to water stress

Pierce’s Disease Oleander Leaf Scorch Coffee Leaf Scorch Almond Leaf Scorch

Maple Leaf Scorch Maple Leaf Scorch Elm Leaf Scorch Oak Leaf Scorch

Phony Peach Fruit No leaf scorching CVC fruit and stunted growth No leaf Scorching

Pierce’s Disease Symptoms Leaf scorch symptom 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

PD Threatens Other Crop Industries 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

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

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

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

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 .

Recombinase-based marker excision system => GM Vitis marker free

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

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

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

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

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?

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?

Potential Interrogations of a GM Grapevines or Rootstocks What are the data requirements?

Potential impact from gene introgression

Potential Weediness

Potential impact on non-target organisms

Transmission of molecules across graft unions