Modern Agricultural Biotechnology: Progress in genetic improvement of plum

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1 Modern Agricultural Biotechnology: Progress in genetic improvement of plum Ralph Scorza USDA-ARS Appalachian Fruit Research Station Kearneysville, West Virginia

2 World-Wide Adoption of Genetically Engineered Crops Soybean, Maize, Canola, Cotton

3 Sugarbeet Biotech herbicide tolerant sugarbeet was grown on 0.5 million hectares in the USA and Canada at 97% adoption rate. Alfalfa 425,000 hectares of biotech RR alfalfa was planted in the US in Poplar Biotech Bt poplar was grown on ~500 hectares in China. Papaya Biotech PRSV resistant papaya was planted in Hawaii (2,000 hectares with a 60% adoption) and in China at 6,275 hectares. Squash 2,000 hectares of biotech virus resistant squash was grown in the US.

4 Biotech crop Facts million - Farmers planting biotech crops - 29 countries - planting biotech crops - 18 Years - Farmers have planted biotech crops M hectares - Global biotech crop area - $44 billion - Increase in net income for farmers - 1 trillion - Estimated number of meals consumed with biotech ingredients Reliably documented human or animal safety issues ISAAA report 2011

5 We all can benefit from the use of biotech crops: Farmer benefits increasing yield improving protection from insects and disease increasing their crops tolerance to heat, drought, and other environmental stress Processor benefits improve the quality of animal feed, food and energy sources. Consumer benefits Increased protein Healthier oils Reduced exposure to agricultural chemicals Environmental benefits Conserve soil and energy Reduce greenhouse gases Minimize use of toxic chemicals Conserve soil fertility and natural resources Reduce need for new agricultural land

6 Biotech crop benefits -1.9 billion kilograms of reduced carbon dioxide emissions billion kilograms of carbon sequestered in the soil -473 million kilogram reduction in pesticides applied from 1996 to million hectares of land preserved between 1996 and million tons of additional food, feed and fiber produced from 1996 to 2011 (ISAAA report 2013)

7 Some potential environmental benefits for 50% adoption of the currently available biotech crops in the EU - pesticide use would decrease by 14.5 million kg /year - reduction of 7.5 million ha sprayed million litres of diesel saved - reduction of 73,000 tons of carbon dioxide being released into the atmosphere. Park and Phipps (2002) Journal of Animal and Feed Sciences, 11:1-18.

8 HoneySweet Plum Resistant to Sharka: A Potential Benefit to European agriculture - Co-developed by U.S. and European partners - Field tested in Europe for over 15 years - Solely the work of publicly-supported scientists, at public research institutions For the benefit of growers and consumers

9 The need for PPV resistance PPV Symptoms -Fruit deformation and reduced quality -Premature fruit drop -Leaf chlorosis -Tree decline in severe infections

10 PPV infection in selected European countries: Serbia - Croatia 42 million bearing plum trees; 58% are infected with PPV survey found 51% of sampled plum trees infected Bosnia-Herzegovina - up to 41% of plum trees infected. Bulgaria - infection in plums 62% Romania infection in plums 70% Moldova yield loses in plum 16-48% Czech Republic 30 years average fruit yield dropped by 80% and the number of plum trees has been reduced from 18 million to 4 million. Greece apricot production decreased from 35% to 13% of world production due to PPV Spain million PPV-infected trees removed between 1989 and 2006 at a cost of over 63 M Euros 2006 OEPP/EPPO 36 (2) Zagrai et al UASMV

11 Development of HoneySweet PPV resistant plum variety Gene discovery, Vector construction, Transformation Plant establishment, Propagation, Greenhouse testing Field testing U.S Field testing Europe and continues today Research and regulatory data accumulation (Over 40 publications from this work) 14 Years

12 Development of HoneySweet PPV resistant plum variety R&D 14 Years Regulatory approval for HoneySweet 7 Years Full Regulatory Approval in U.S. 2011

13 U.S. Regulatory Approval Three agencies: Jurisdiction Safety for Agriculture Safety for Food Safety for Environment

14 Examples of some of the information provided to U.S. regulators: Transformation system Donor genes and sequences Molecular characterization (DNA, RNA, protein) Mechanism of resistance Stability of resistance Inheritance of resistance Gene flow Fruit compositional analyses Allergenicity potential Environmental consequences

15 European colleagues have worked together with The U.S. team since In 1996 field tests were initiated In Poland, Romania, Spain, and in 2002 in the Czech Republic. These data were included in the U.S. regulatory dossier.

16 How effective is HoneySweet? In over 15 years of field testing no HoneySweet trees have been naturally Infected with PPV by aphids European HoneySweet field test harvest

17 % PPV Temporal spread of Plum pox virus - Romania conventional years C5

18 Graft inoculation of HoneySweet with PPV PPV infected plum tree HoneySweet tree PPV infected bud

19 Czech Republic Field Test - Graft inoculation HoneySweet graft inoculum (PPV-Rec) Year 2 Year 5 Polak et al., 2008

20 mg per 100 dry grams mg GAE/Kg dry weight % by dry weight HoneySweet safety studies FRUIT COMPOSITION STUDIES 70 Average Total Sugars Stanley JoJo HoneySweet Others Average Phenolics 7000 Stanley Jojo HoneySweet 3000 Others Average Vitamin C Stanley JoJo 30 HoneySweet 20 Others 10 0

21 Molecular evaluations: - the entire genome of HoneySweet was sequenced - no genes interrupted, no effect of insert on flanking genes - all RNAs were sequenced - no unexpected RNAs produced - HoneySweet produced less PPV-CP RNAs than produced in PPV infected plums - no unexpected proteins produced

22 Results of Environmental Risk Studies Zagrai et al Transgenic plums expressing Plum pox virus coat protein gene do not assist the development of virus recombinants under field conditions. Journal of Plant Pathology No virus recombinants Capote et al Risk assessment of the field release of transgenic European plums susceptible and resistant to Plum pox virus ITEA 2007 No effects on non-target insects Capote et al Assessment of the diversity and dynamics of Plum pox virus and aphid populations in transgenic European Plums under Mediterranean conditions Transgenic Research 2008 No effects on aphid populations No effects on virus diversity Zagrai et al Plum pox virus silencing of C5 transgenic plums is stable under challenge inoculation with heterologous viruses. Journal of Plant Pathology No breakdown of resistance in presence of other Prunus viruses Gene flow is low (Scorza et al PLOS ONE in press)

23 HoneySweet It is highly resistant to PPV. No trees were ever infected by aphids. Resistance to all strains tested Fruit quality is excellent The transgene and resistance could be transferred to progeny through traditional cross hybridization and the resistant seedlings could be readily identified. HoneySweet may be useful as a PPV resistant rootstock Approved for cultivation and consumption in the U.S.

24 This work is a U.S. - European collaboration M. Ravelonandro, France M. Cambra, Spain N. Capote, Spain T. Malinowski, Poland I. Zagrai, Romania J. Polak, Czech Republic J. Kundu, Czech Republic I. Kamenova, Bulgaria A. Atanassov, Bulgaria S. Paunovic, Serbia S. Dolgov, Russia Shouldn t European growers and consumers also benefit? Black Sea Biotechnology Association R. Scorza, USDA A.M. Callahan, USDA C. Dardick, USDA and others. HoneySweet International Working Group

25 HoneySweet is being made freely available in the U.S. Those outside the U.S. wishing to exploit HoneySweet will be responsible for obtaining the required regulatory clearances. Qualities- Sweet (21% sugar), large size (60 g), productivity very good. A good variety for the fresh market. Brandy, marmalade, drying not yet tested.

26 Scientists at the Crop Research Institute, Czech Republic, are developing a dossier to submit to EFSA Based on over 15 years of study in Europe and the U.S. Czech Republic present Poland Romania , Spain 1996-present

27 This technology can help solve the problems that will be facing Europe, the U.S. and the rest of the world including climate change, the spread of invasive species, and population pressures for increased food production. Wisely used, genetic engineering can be an important tool to help meet these challenges. HoneySweet International Working Group

28 Scientific Institutions expressing positive positions on GMO Crops Academy of Medicine, France American Medical Association American Society for Plant Biology Brazilian Academy of Science Chinese Academy of Science Food and Agriculture Organization Indian National Science Academy Mexican Academy of Science Pontifical Academy of Sciences Romanian Academy Romanian Academy of Agricultural and Forestry Sciences Royal Society of London Third World Academy of Science U.S. Department of Agriculture U.S. National Academy of Sciences U.S. National Research Council World Health Organization and others.