Agricultural Biotechnology: prospects and problems.

1. Agricultural Biotechnology: prospects and problems. Piero Morandini Dept. of Biology University of Milan (Italy)

2. Outline • Present transgenic crops (compared to what?) • Problems (risk vs. benefits) • Future prospects • Lies, dammed lies and statistics

3. Compared with what? • It is impossible to understand and judge ag-biotechnology without knowing the tools of breeding (how crops are created) • Thousands of new crop varieties have been created using less precise techniques • Conventional breeding (mutation breeding, embryo rescue, wide area crosses, double haploid...) does manipulate genes

4. Triticale – a new crop plant species durum wheat Triticum turgidum rye Secale cereale triticale X Triticosecale Hybridize X Embryo culture & chromosome duplication to restore fertility Breeders have combined durum wheat and rye to produce triticale, which has a larger gene pool than is present in either of the parent crops. photos compliments of Calvin Qualset, UC Davis

5. Is triticale natural? 1876A. S. Wilson a schottish botanist crosses what and rye. The result is a new sterile plant. 1937using colchicine, researchers double the chromosome number (and of genes as well) 1970-80First commercial triticale varieties are released 2010More than 4 Mha are being cultivated

6. How many... • ... silent genes coding for allergens or toxins have been activated in triticale? • ... endogenous genes have been disrupted in their action? • ...safety tests on target and non target organisms have been conducted in Europe before releasing triticale? Any fear or objection raised for a transgenic plants MUST BE RAISED also for varieties like triticale (& thousands of others)

7. Herbicide tolerant Sunflower Turkish Sunflower production in 2009 was about 1 Million t Total crop surface 580.000 ha  HT hybrid 30% • Cross-breeding to wild relatives – YES • Contamination of sensitive varieties – YES • Uses more herbicide – YES; • Imazamox is more persistent in soil. Better avoid planting sensitive varieties 21 moths after first use. • Spontaneous occurrence of resistance is common after 4 o 5 treatments. Increases herbicide use. • Invasive – YES , Sunflower has still some weedy characters (persistence in fields and outside fields). Stable populations are known in the wild. main oil crop in Turkey Obtained by conventional or mutation breeding. It does NOT undergo the same extensive testing for environmental / health safety as transgenic varieties Also: maize, rice, wheat, oilseed rape, lentil, soybean… Sala et al (2008) Theor Appl Genet 118:105-112 http://www.ars.usda.gov/is/AR/archive/jan05/sunflower0105.htm

8. mutagenesis Herbicide tolerant sunflower Herbicide tolerant sunflower spont. mutation transgenesis Any loss of function… Two methods, one result Sunflower Same result using different methods:  is one evil and the others benign? Focus on product risks, not on method used * Allergen inactivation (soybean) * Toxin inactivation (cotton ) *... Morandini (2010) New Biotechnol.

9. Allergies caused by plants Eight foods account for 90% of all food-allergic reactions. milk, egg, fish, shellfish, peanut, tree nut, soy, wheat Pollen is the major cause of respiratory allergy. At least 40% of type 1 allergic patients are sensitized against grass pollen allergens • Contrary to common perception, transgenic plants never caused allergic reactions to consumers. Many conventional crops do it regularly • If a gene used for transgenesis comes from a plant containing allergens, the transgene is checked for allergenicity

10. Work being done on plant allergens: Transgenesis, rather than a cause of allergy… … can be part of the solution • Apple • Peanut • Wheat (celiac disease) • Soybean • Ryegrass • Birch

11. Soybean allergen: P34 US/Europe: 5 - 8% of babies and 2% of adults allergic* to soybeans Dominant soybean allergens is P34: > 65% of soy-sensitive patients react only to P34 protein Transgenic soybean without P34 published in 2003 (Herman et al., 2003) Herman et al., (2003) Plant Physiol. 132:36-43 No difference in composition, development, structure, or ultrastructure when compared with control plants. No other significant changes in polypeptide pattern

12. “Regulatory difficulties and the lack of acceptance of GM soybeans by the baby food and formula industry makes using such an allergen-suppressed soybean difficult [read impossible] at the present time.” Alternative approach: identify soybeans with little/no allergen  screen the USDA soybean germplasm collection Joseph et al. (2006) 46:1755-63 Out of > 16 266 accessions soybean germplasm screened, 12 lines (2 in the cultivated soybean) have no P34 allergen Why these two soybean plants lack the antigen?

13. Environmental risks  a nice character (for us) Wild oat: at maturity seeds are no longer present in wild species Cultivated oat: at maturity seeds are retained; a strong treatment is needed for release (all cereals and in general for crop plants)

14. Wild wheat – cultivated wheat

15. Wild plants withstand difficult conditions

16. Crop plants are weak Potato is devoured by the Colorado potato beetle Crop plants have difficulties at surviving in natural environments

17. This requires a lot of labour and some technology Spontaneous transition If crop plants were natural... ...agriculture would not be a problem!

18. Problems (risk vs. benefits) Are there risks in transgenics? Of course there are, but no different from naturally occurring or conventionally bred plants. * Insisting on ‘environmental risks’ without considering the intrinsic weakness of crops * Insisting on ‘potential’ healts dangers of transgenic food without considering real (similar or worse) from conventional or organic * Non considering benefits IS INSANE!

19. Risk: red rice and HT rice(overlooked dangers) Cecilia • Weedy rice (aka ‘red rice’) is closely related to cultivated rice. • At seedling stage, weedy plants are difficult to distinguish from the crop. • At maturity, red rice is taller. • Infestations are reported for 40-75 % of the rice area in Europe. Red rice Cultivated rice

20. Seeds fall before harvest - may stay dormant 2-5 yr • 11 plants m-2 means a yield loss of about 40% Cultivated rice Red rice • In short: dangers for the environment are essentially of agronomic type (damage in fields, not in the wild) • FOOD SECURITY !!! Example (II): Bt corn Transgenic –herbicide resistant rice: excellent control of red rice for 2-3 yrs Resistant gene introgressed into weedy rice  then technology is lost * A problem in rice growing areas with direct seeding * Herbicides that kill red rice also kill cultivated rice

21. An overwhelming consensus • Many major national and international Academy of sciences (United States, India, Brazil, France, Germany, United Kingdom, Italy, India, China, Mexico, Pontifical Academy of Science and the Third World Academy) have issued positive statements on this technology. • Many scientific societies international organizations (WHO, FAO…)* concluded that GE crops present no new or different risks, and can (and in fact, do) reduce or ameliorate some of the negative impacts of conventional agriculture. * for a long, but incomplete list, see: http://users.unimi.it/morandin/Sources-Academies-societies.doc

22. Future prospects What can Ag-biotech do for Turkey (Italy, Europe or the Third word)? One example from western countries  "pest resistant" celery (many, many more available: e.g. apple scab, increase productivity in rapeseed, drought tolerance… One example for the third world:  Edible cotton seed (improved Cassava, Golden rice and other golden crops...).

23. 1 2 3 4 sepal petal stamen carpel Refreshing the vocabulary

24. Identity crisis Mutant: Apetala Petal  flower Sepal  leaf

25. More problems Pistillata Apetala3 Agamous petal  sepal petal  sepal stamen  petal stamen  carpel stami  carpelli carpel  flower Images courtesy of L. Colombo (Unimi)

26. and Cauliflower… …as well as Arabidopsis

27. Even cucumbers have problems One of the most aggressive insects damaging cucumbers in greenhouses in Canada & USA Western flower thrips Frankliniella occidentalis Female cucumber flower Images courtesy of M. Kater (Unimi) http://bugguide.net/node/view/252440/bgimage

28. Female flower • meeting place • mating place Life cycle of Frankliniella occidentalis egg Adult larva Pupa Images courtesy of M. Kater (Unimi)

29. Mutante Green petals: B (CUM26) B A C S S --- C 1 2 3 4 cum26 Petal  sepal Images courtesy of M. Kater (Unimi)

30. 200 200 100 100 Apical leaves Basal leaves 8000 8000 total damage mm2 total damage mm2 thrips larvae thrips larvae 0 0 4000 4000 0 0 C D wt gp mutant wt gp mutant Green petals mutant Take home message: genetic engineering is a more predictable, more precise, faster and less expensive method to introduce a character Wild-type Images courtesy of M. Kater (Unimi)

31. Gossypol, a problem • Cotton produces 1.65 kg of seed for 1 kg of fiber • Due to gossypol, a cardio- and hepatotoxic terpenoid  seed unfit for consumption by humans and monogastric animals) • Seed contains 21% oil and 23% high-quality protein Sunilkumar et al. (2006) P.N.A.S. 103:18054–18059

32. Cottonseed may help feed the world • Used as feed for ruminant animals (whole seeds or meal after oil extraction) • 44 million metric tons (Mt) of cottonseed (9.4 Mt of protein) • Could fulfill protein requirements of half a billion people each year (50 g/day rate) Sunilkumar et al. (2006) PNAS 103:18054-9

33. RNAi inactivating δ-cadinene synthase in the seed Chlorophyll Carotenoids … Proposed biosynthetic pathway of gossypol Sunilkumar et al. (2006) PNAS 103:18054-9

34.  a strong reduction of gossypol in seed Levels of gossypol (mg/mg seed) for each individual seed Sunilkumar et al. (2006) PNAS 103:18054-9

35. Transgenic vs. conventional • A glandless mutant was obtained with conventional strategies. Varieties with this trait were a failure under field conditions (extraordinarily susceptible to a host of insect pests) • Terpenoids protect the plant from both insects and pathogens • The transgenic approach achieved a goal classical breeding was unable to obtain (specific reduction in seed) Targeted gene silencing can be used to modulate biosynthetic pathways in a specific tissue to obtain a desired phenotype. Impossible by traditional breeding  Texas A&M University and U.S. Department of Agriculture Sunilkumar et al. (2006) PNAS 103:18054–18059

36. Overegulation is killing “[this] approach…not only improves food safety but also provides an additional and potentially extraordinary mean to meet the nutritional requirements of the growing world population without having to increase either crop yields or acreage planted” (Sunilkumar et al., 2006) “Our hope is to get through regulatory approval process in the U.S. first. However, it takes $50-100 million to go through the process. At this point, we don't know where the money is going to come from, but we are exploring various possibilities. Getting U.S. approval will make it easier to then get permit in other countries. We will be especially interested in some African countries and some Asian countries. “ (personal communication by Keerti S. Rathore) Gene technology could improve food safety, food security and reduce environmental impact. Regulation is a major obstacle

37. Before cultivation is allowed, a scientific dossier must be produced Transgenic variety Conventional variety Images cortesy of A. McHughen (Canada)

38. Collecting cases with PRRI  a book (please, help!) http://www.flickr.com/photos/26138378@N03/2630688669/ www.sardi.sa.gov.au/__data/assets/image/0008/45458/peaweevil_damage1.jpg

39. Turkey must decide: • use biotech to improve agriculture or • - sit and wait for collapse http://callmeshree.files.wordpress.com/2010/07/damned-if-you-do-dont.jpg

40. Lies, dammed lies and statistics • Several myths about transgenic plants • sterile • unnatural • inherently (more) dangerous • will destroy biodiversity and/or local landraces • will make poor farmers even poorer “Europe doen’t want / need transgenics” “Introduction of transgenics destroys biodiversity” • Italy imports 4 Mt of soybean and derivatives to make high value products (and Europe imports 50 Mt)

41. Question:How many transgenic corn varieties are cultivated in Europe now? * 100 varieties of Bt maize in cultivation in Spain (starting from one)* More than 600 varieties of Bt cotton in India" The number of Bt hybrids released for commercial cultivation till date has crossed 600 with more than 35 seed companies and public sector institutions currently engaged in their development"* More than 400 varieties of RR soybean in Argentina* Probably more than 1000 varieties of RR soybean in the US • Conclusion: there are many lies around  find reliable sources Why is Europe not cultivating transgenics?

42. EU spent 200 M€ for 130 research projects (mainly biosafety)  No negative results reported Truth is: disconnection between science and politics The technology has great benefits and a much greater potential, if... - accept some (minute) level of risk - regulation becomes more sensible

43. http://www.poppyseedtea.com/ Poppy seed tea can kill you

45. The P38 and the apple A true desire, when does not come to terms with reality, but follows the path of irrational utopia, turns into lie and can only end up in murderous madness and self destruction. S. Allevato e P. Cerocchi (2009) “La P38 e la mela”, publ. ITACA, p.173 Un desiderio vero, quando non fa i conti con la realtà, ma imbocca la strada dell’utopia irrazionale, diventa menzogna, e non può che condurre alla follia omicida e all’autodistruzione. S. Allevato e P. Cerocchi (2009) “La P38 e la mela”, ed. ITACA, p.173

47. Are transgenic plants not natural? The most common method to make them exploits bacteria (natural transgenics) Transgenic callus on a oleander leaf (contains bacterial genes)

48. Transgenic seed exhibits a large reduction in Gossypol level. A monogenic trait: the reduced gossypol trait cosegregates with the transgene  Much more predictable, stable, specific Sunilkumar et al. (2006) PNAS 103:18054–18059