Do Russia and Eastern Europe need GM plants

Do Russia and Eastern Europe need GM plants? Konstantin Skryabin Centre “Bioengineering”, RAS

Acreage planted, 2008: USA > 100 000 ha Russia > 80 000 ha Kazakhstan > 15 000 ha Ukraine > 15 000 ha

Adverse climatic conditions LOW TEMPERATURE Russian Federation DRYNESS Kazakhstan

Crop losses in Russian Federation, 2008 Crops Gross yield, weeds pests viruses Total losses, thous. tons % % 63800 11484 18 8294 13 4466 7 31 Sunflower 7400 1406 19 - - 38 Sugar beet 29000 7830 27 3190 11 2900 10 48 Potatoes 28800 4608 16 3456 12 4320 15 43 6700 1340 20 1005 15 - - 35 752 128 17 150. 4 20 - - 37 Wheat Corn (for grain) Canola thousand tons

Crop losses in Ukraine, 2008 Crops Gross yield, thousand tons Total from pests and viruses % losses, thousand tons 25882. 6 30 7765 Sunflower (for seed) 6520. 3 38 2478 Sugar beet 13724. 7 48 6588 Potatoes 19545. 4 43 8405 Corn (for grain) 11422. 3 35 3998 2872. 9 37 1063 Wheat Canola

Crop losses in Kazakhstan Republic, 2007 Crops Gross yield, thousand tons Total from pests and viruses % losses, thousand tons 20 099 20 -30 4020 – 6030 Sunflower 208 4 -39 8 – 81 Sugar beet 311. 3 10 -20 31 – 62 2 402. 4 20 -70 480 – 1682 442 10 -20 44 – 88 Wheat Potatoes Raw cotton

Total crop losses in three countries, 2008 Gross yield, thousand tons % losses, thousand tons 109 781. 6 29 31 837 Sunflower 14 128. 3 33 4 662 Sugar beet 43 036 37 15 923 Potatoes 50 747. 8 44 22 329 Corn 18 122. 3 35 6 343 3624. 9 37 1 341 442 15 66 Crops Wheat Canola Raw cotton Total

Development of Russian potato varieties, resistant to Colorado beetle

Field performance of transgenic potato, resistant to Colorado beetle Nevsky, GM modified Lugovskoy, GM modified Control plants Plot, certified by IAC GEA, South Russia, Krasnodar, 2002

Transgenic potato, resistant to Colorado beetle variety Lugovskoy, cry 3 A GM modified Plot, certified by IAC GEA, Russia, Moscow region - 2002

UPOV ESTIMATION OF CHARACTERISTICS OF GM POTATO LINES Plot, certified by IAC GEA, Moscow region - 2002

Transgenic potato obtaining Southern-blot analysis Border identifiers for Bt-potato X+A sequences for Lugovskoy+ Elizaveta+ and Nevskiy+ were used in patent as unique border identifiers

PRODUCTION OF HIGH QUALITY POTATO MINI-TUBERS

GM potato IPR protection

Food certificates # 77. 99. 26. 11. У. 6088. 7. 06 (07 July 2006) Lugovskoy 1210 amk event # 77. 99. 11. У. 14145. 12. 05 (14 December 2005) Elizaveta 2904/1 kgs event Patents # AND DATE OF PRIORITY THE REGISTRATION DATE ISSUE # 2231550 24 April 2002 # 2231549 24 April 2002 27 June 2004 obtain GM plants of Lugovskoy variety 27 June 2004 obtain GM plants of Nevskiy variety # 2231251 27 June 2004 # 2286385 23 November 2004 27 June 2004 obtain GM plants of Elizaveta variety 27 October 2006 Recombinant DNA sequence including Elizaveta variety genome plant DNA and transgenic insert # 2286386 14 February 2005 27 October 2006 Recombinant DNA sequence including Nevskiy variety genome plant DNA and transgenic insert # 2007110779 26 March 2007 Recombinant DNA sequence including Lugovskoy variety genome plant DNA and transgenic insert

Development of Russian sugar beet varieties, resistant to viral infection ü viral resistance ü herbicide resistance

Beet yellows virus (BYV) Total crop weight loss caused by BYV is up to 40%, with sugar reduction of 8 -10%

Beet necrotic yellow vein virus (BNYVV) BNYVV infections may cause reduction of crop weight and sugar content of up to 50 -70% Rhizomania distributed over most sugar beet growing areas in the world: Asia, China, Japan, the USA, Europe countries and Russia

Development of strategy for pathogen-derived resistance in Nicotiana benthamiana to Beet yellows virus and Beet necrotic yellow vein virus BNYVV infections may cause reduction of crop weight and sugar content of up to 50 -70% and 5 -10%, respectively Total crop weight loss caused by BYV is up to 40%, with sugar reduction of 8 -10% Validation of the approach target for silencing + PTGS inducer target for silencing Co-expression of the target and the inducer of PTGS 10 times reduced GFP in comparison with the target only expression

Validation of the approach B А p. NR_AMVGFP_BNYVVsil (target for silencing) + p. BI_BAR_BNYVVsil (PTGS inducer) Leaf 1 GFP Leaf 2 Leaf 3 А B А B Western-blot analysis of GFP expression Co-expression of the target and the inducer of PTGS 10 times reduced GFP in comparison with the target only expression

Transgenic sugar beet obtaining

MF 3 - peptidyl-prolyl cis-trans isomerase, FKBP-type, from Pseudomonas fluorescence strain 197 MW 17. 6 k. D mliaankavs idytltndag evidssagga plvylqgagn iipglekale gkqvgdelkv svepedayge yaaelvstls rsmfegvdel evgmqfhasa pdgqmqivti rdldgddvtv dgnhplagqr lnfqvkivai rdasqeeiah ghvhgegghh h

Elicitor activity of MF 3 -protein tested on wheat leaves of variety Mironovskaya 808 against fungus Septoria nodorum MF 3 -protein 1. 25 mg/ml Reference (water)

Elicitor activity of MF 3 -protein tested on wheat leaves of variety Mironovskaya 808 against fungus Puccinia graminis MF 3 -protein 1. 25 mg/ml Reference (water)

Resistance of transgenic potato plants’ tubers to Erwinia carotovora Potato tuber pieces of non-transgenic variety Nevsky Potato tuber pieces of transgenic variety Nevsky line 56 Potato tuber pieces, place in a Petri dish on moist sterile filter paper, and inoculate with 0. 1 - 1 of the bacterial suspen-sion (ca. 106 CFU/ml). Incubate at 20 -27°C for 48 h and probe the tissue surrounding the inoculation site with a spatula to determine whether decay and tissue maceration has occurred.

Resistance of transgenic potato tubers to bacterial phytopathogen Erwinia carotovora Lines of Nevsky potato variety Diameter of necrotic lesions, mm Non-transgenic N 53 transgenic N 56 transgenic N 71 21. 9 НСР 0, 05 3. 5 14. 1 6. 9 9. 4

Sensitivity of transgenic potato plants to the late blight disease under natural infections conditions in the field Results of testing transgenic potato plants on PVX resistance under natural infection conditions

Resistance of transgenic Canola to Plasmodiophora brassicae. 1 а/7, Т 25/8, Т 25/12, 1 а/3, Т 25/9 - transgenic lines canola variety Westar. Т 32/7 - transgenic line of Cauliflower Fraction of diseased plants

Development of salt–tolerant plants

Possible H+ -pyrophosphatase localization in plant cell Antiporter Н+ATPase Н+ Н+ATPase Na+ antiporter PP Н+ vacuole tonoplast PP Н+ Н+ cytoplasmalemma PP - pyrophoshpatase from Rhodospirillum rubrum

Salt tolerance of transgenic tobacco expressing H+ pyrophosphatase gene from Rhodospirillum rubrum Control plants Na. Cl 25 m. M 175 m. M Seed germination efficiency at different Na. Cl concentrations Transgenic plants Growth of control and transgenic plants at different Na. Cl concentrations Control Plants, 150 m. M Na. Cl Transgenic Plants, m. M Na. Cl 150

Evolution of the regulatory framework in Russia (genetic engineering) 1996 Federal act "On State regulation of genetic engineering activity" 2000 Amendments in respect of gene therapy and genetic diagnostics to Federal act "On State regulation of genetic engineering activity" 2000 Instruction by the RF Chief State Sanitary Physician : "On the order of hygienic expertise of food products, derived from GMOs” 2001 Government Act “On state registration of GMOs”, Non upgraded till now 2002 Government Act “On state registration of GM feed”, upgraded 2006 2002 “On protection of natural environment” (2002 ) 2003 The order by the RF Chief State Sanitary Physician : “On the order of microbiology molecular-genetic expertise of GM microorganisms, used in food production” 2005 Federal act “On protection of consumer’s rights”, amended in 2007, 2008 2004 - Ongoing technical regulation reform. Ongoing administrative reform. 2009

Major points of GMOs Regulation Systems in Russian Federation Food safety assessment GMOs safety assessment Biosafety trials + ? Feed safety assessment Federal Service for Supervision of Consumer Rights Protection and Human Welfare Federal Service for Veterinary Surveillance GM Plant Food Registration Variety Trials N 0 events 15 events Feed Registration 11 events (168 GM products) Placing on the market Ecological Expertise Pesticide Registration GM CROP Commercial growing NO events

Russian Federation: Biotech food labeling “Labeling law” (2002, 2007) • First step: September 2002 - Sanitary and Epidemiol. Rules 2. 3. 2 1078 -01 2002 – Mandatory labeling. 5% threshold. • Second step: Sept 2007 - Sanitary and Epidemiol. Rules • Mandatory labeling for product consists of, contains or is produced from GMOs. 0, 9% threshold (ingredients considered individually) • Mandatory labeling for product is produced from GMOs irrespective of the detectability of DNA • Adventitious presence if transformation events are no more than 0, 9% weight

Dynamics of GM foods and feeds registration Number of registered transformation events/year

Russian Federation: GM food certification (15) PLANT TRAIT COMPANY YEAR Potato, variety “Elizaveta” colorado potato beetle resistance Centre “Bioengineering”, RAS, Russia 2005, for unlimited period Potato, variety “Lugovskoy” colorado potato beetle resistance Centre “Bioengineering”, RAS, Russia 2005, for unlimited period Soybean А 2704 -12 “Liberty Link” glufosinate ammonium tolerance Bayer Crop Sciences 2002 -2007 extended for 2012 Soybean А 5547 -127 “Liberty Link” glufosinate ammonium tolerance Bayer Crop Sciences 2002 -2007 extended for unlimited period Soybean GTS 40 -3 -2 glyphosate tolerance Monsanto Company, USA 1999 -2002, extended for unlimited period Maize MON 88017 resistance to CRW (Coleopteran, Diabrotica sp. ) and glufosinate ammonium tolerance Monsanto Company, USA 2007, for unlimited period

Russian Federation: GM food certification PLANT TRAIT COMPANY YEAR Maize Roundup Ready® GA-21 glyphosate herbicide tolerance Monsanto Company, USA; from 2007 Syngenta Seeds, Inc. 2000 -2003, extended; from 2007 for unlimited period Maize Yieldgard® MON-810 resistance to European corn borer Monsanto Company, USA 2000 -2003, extended for 2009 Maize Roundup Ready® NK 603 glyphosate herbicide tolerance Monsanto Company, USA 2002 -2007 extended for unlimited period Maize MON 863 resistance to corn root worm (Coleopteran, Diabrotica sp. ) Monsanto Company, USA 2003 -2008 extended for unlimited period Maize Liberty-Link™ Т-25 glufosinate ammonium tolerance Bayer Crop. Science (Aventis Crop. Science) 2001 -2006, extended for unlimited period Maize Bt-11 resistance to European corn borer and glufosinate ammonium tolerance Syngenta Seeds, Inc. 2003 -2008 extended for unlimited period

Russian Federation: GM food certification PLANT TRAIT COMPANY YEAR Maize MIR 604 resistance to corn root worm (Coleopteran, Diabrotica sp. ) Syngenta Seeds, Inc 2007, for unlimited period Rice LL 62 glufosinate ammonium tolerance Bayer Crop Sciences 2003 -2008, submitted for extending Roundup Ready® sugar beet H 7 -1 glyphosate herbicide tolerance Monsanto Company/ KWS 2006, for unlimited period T O T A L: 15 transformation events

Russian Federation: GM feed certification Plant/f eed company GMO year Soybean Monsanto Co. soybean GTS 40 -3 -2 2003 – 2008 extended for 2013 Soybean Bayer Crop. Sciences soybean А 2704 -12 2007 - 2012 Soybean Bayer Crop. Sciences soybean A 5547 -127 2007 -2012 Maize Monsanto Co. maize GA 21 2003 – 2008 extended for 2013 Maize Monsanto Co. maize NK 603 2003 - 2008 extended for 2013 Maize Monsanto Co. maize MON 810 2003 - 2008 extended for 2013 Maize Monsanto Co. maize MON 863 2003 - 2008 extended for 2013 Maize Monsanto Co. maize MON 88017 2008 -2013 Maize Syngenta Crop Protection AG maize MIR 604 2008 -2013 Maize Syngenta Crop Protection AG maize Bt 11 2006 - 2011 Maize Bayer Crop. Science Gmb. H maize T 25 2006 - 2011 T O T A L: 11 transformation events and 168 Feeds derived from these plants

Russian Federation-Ukraine-Kazakhstan The Cartagena Protocol on Biosafety to the Convention on Biological Diversity Country Ratification status Russia No Ukraine Accession, 2002 -12 -06 Kazakhstan Accession, 2008 -09 -08 Legislation Ukraine Kazakhstan Law “On State system of biosafety in Creating, testing, transporting and using of GMOs” № 1103 -V, 05/31/2007 (approved) Government Act “Rules for circulation of genetic modified objects”, 06/27/2008 (approved)

“Caution! Scorpion’s Gene” Doc movie 01’ 11” April, 9 2009 TV-Channel “Russia” Mikhail Bobrovnik Oleg Vetviсky All-Russian State TV and Broadcasting Company 2009

01’ 20” Svetlana Ryzhakova (Moscow) is raising a daughter, Christina. A year ago, when they were coming home from a Middle East country (their dad works under a contract there), the fearful acute attack of a disease has began with the little girl directly in the plane: the temperature rose, the girl had become short of breath, and she began convulsing.

01’ 44” 02’ 02” 01’ 59” Doctors in Moscow hospital detected the very heavy allergic reaction in a little patients, but for what? Baby held several days in an intensive care before doctors were able to understand what has caused the attack.

02’ 43” 02’ 05” Purchased overseas babies formula has included an additive which was banned in the Russia - a genetic modified soya protein. However the same genetics achievements have hardly killed Kristina. A rare allergic reaction specifically for GM soyabean protein was found in the little girl.

10’ 17” 10’ 20” After transgenic tomatoes, wheat and potatoes with scorpion gene introduced into crops, have been marketed. Manufacturers advertise this products as 21 st Centure food. 10’ 22”

21’ 31” In 2003 two babies have died in Israel. The results of investigations have shocked as researchers, as parents: it was proved that the infants died from the lack of vitamin B 1, which they had to get out of the babies formula.

21’ 12” “It was suggested that so-called «supersoya» (which, of course, has been genetically modified!) would be a source of vitamin B 1. BUT, it wasn’t vitamin B 1 in this soybean! During a designing process «something» has happened, and the normal pathway of vitamin B 1 biosynthesis has been blocked, that resulted in the absence of the product. Plant synthesizes a few tens of thousands of different compounds, and absolutely impossible to predict what will not be? or what will be? or would it work strongly or slowly? »

30’ 45” 30’ 49” 30’ 41” Once more argument of anti-GMO activists in favor of complete GMO banning is the EU position on this issue. Transgenes are the rarities on supermarket shelves of European cities, and in 1998 EU countries introduced de facto moratorium on the importation of GM products from the United States at all.

41’ 50” So what is the transgenes? Product of the XXI Century or a delay action «food bomb» ? Wheat with scorpion gene. Tomato with shark gene. Maize with monkey gene.