L’insostenibile leggerezza della regolamentazione delle piante transgeniche

1. Il costo della non scienza in agricoltura Il rigetto delle colture transgeniche in Italia e in Europa The cost of fiction vs. science in agriculture The rejection of transgenic crops in Italy and Europe 12 Giugno 2014 L’insostenibile leggerezza della regolamentazione delle piante transgeniche The unbearable lightness of the regulation of transgenic plants Piero Morandini Dipartimento di Bioscienze Università di Milano

2. Sommario • GMO or non-GMO: what do you mean, sir? • Process-based regulation: is it reasonable? • Risks: compared to what? • The unbearable lightness: the Italian version • Exit strategies

3. GMO o non GMO? Mature spikelets in wildoat: seedsfall to the ground in a fewdays Mature spikelets in cultivatedoat: seeds stay attached to the spikelets. A harsh treatment (threshing) isnecessary to detachthem.

4. coltivato: ...TCATGAAATGT... selvatico:...TCATGCAATGT... Promotore Regione trascritta Difference? 1 gene over 30.000 Difference in the gene? 1 letter over 15.000 Ideallyone base change over the entiregenome(300 millionbases)isenoughmakereproductionnearlyimpossible in naturalconditions

5. For thousandsyears man selectedfavourablevariantsamongthoseoffered by nature. The results of the selectionwas the accumulation of severalmutations and the creation of cropplants out of wild ones Wild tomato A modern variety of cultivated tomato

6. Present EU regulation Direttiva 2001/18/CE del Parlamento europeo e del Consiglio del 12 marzo 2001 sull'emissione deliberata nell'ambiente di organismi geneticamente modificati Articolo 2 Definizioni Ai fini della presente direttiva si intende per: 1) … 2) "organismo geneticamente modificato (OGM)", un organismo, diverso da un essere umano, il cui materiale genetico è stato modificato in modo diverso da quanto avviene in natura con l'accoppiamento e/o la ricombinazione genetica naturale. Ai fini della presente definizione: a) una modificazione genetica è ottenuta almeno mediante l'impiego delle tecniche elencate nell'allegato I A, parte 1; b) le tecniche elencate nell'allegato I A, parte 2non sono considerate tecniche che hanno per effetto una modificazione genetica; For the English text of Directive 2001/18/CE see: http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32001L0018

7. TECNICHE DI CUI ALL'ARTICOLO 2, PARAGRAFO 2 - PARTE 1 Le tecniche di modificazione genetica di cui all'articolo 2, paragrafo 2, lettera a), comprendono tra l'altro: 1) tecniche di ricombinazione dell'acido nucleico che comportano la formazione di nuove combinazioni di materiale genetico mediante inserimento in un virus, un plasmide batterico o qualsiasi altro vettore, di molecole di acido nucleico prodotte con qualsiasi mezzo all'esterno di un organismo, nonché la loro incorporazione in un organismo ospite nel quale non compaiono per natura, ma nel quale possono replicarsi in maniera continua; 2) tecniche che comportano l'introduzione diretta in un organismo di materiale ereditabile preparato al suo esterno, tra cui la microiniezione, la macroiniezione e il microincapsulamento; 3) fusione cellulare (inclusa la fusione di protoplasti) o tecniche di ibridazione per la costruzione di cellule vive, che presentano nuove combinazioni di materiale genetico ereditabile, mediante la fusione di due o più cellule, utilizzando metodi non naturali. PARTE 2 Tecniche di cui all'articolo 2, paragrafo 2, lettera b), che non si ritiene producano modificazioni genetiche, a condizione che non comportino l'impiego di molecole di acido nucleico ricombinante o di organismi geneticamente modificati prodotti con tecniche o metodi diversi da quelli esclusi dall'allegato I B: 1) fecondazione in vitro; 2) processi naturali, quali la coniugazione, la trasduzione e la trasformazione; 3) induzione della poliploidia.

8. ALLEGATO I B TECNICHE DI CUI ALL'ARTICOLO 3 Le tecniche o i metodi di modificazione genetica che implicano l'esclusione degli organismi dal campo di applicazione della presente direttiva, a condizione che non comportino l'impiego di molecole di acido nucleico ricombinante o di organismi geneticamente modificati diversi da quelli prodotti mediante una o più tecniche oppure uno o più metodi elencati qui di seguito sono: 1. la mutagenesi; 2. la fusione cellulare (inclusa la fusione di protoplasti) di cellule vegetali di organismi che possono scambiare materiale genetico anche con metodi di riproduzione tradizionali. Present EU regulationisprocess, notproduct-based  ALLEGATO II - PRINCIPI PER LA VALUTAZIONE DEL RISCHIO AMBIENTALE  ALLEGATO III - INFORMAZIONI OBBLIGATORIE PER LA NOTIFICA

9. A scientific dossier is required to apply for a cultivation permit Transgenic variety Conventional variety Cortesy of A. McHughen (Univ. of California, Riverside)

10. Is a process-basedregulationscientifically sound? Test case 1: loss of function mutations Transcription Can be achieved by: -spontaneous mutation -chemical/physical mutagenesis -transposons/T-DNA -directed mutation -RNAi -Talen/ZFN/CRISPR -… Translation Protein toxin ‘good’ phenotype

11. A case of double standards Amflora potato: starch consists almost esclusively of amilopectin. Obtained in the early 90s through antisense against an enzyme involved in starch synthesis (GBSS). First field trials in 1993 (see specific dossier) First application to the EU in 1996. Approved in 2010  15 years to gain approval… Removed from market in 2012 http://www.basf.com/group/corporate/en_GB/news-and-media-relations/science-around-us/amflora/ Proof of concept: Visser RG, Somhorst I, Kuipers GJ, Ruys NJ, Feenstra WJ, Jacobsen E. (1991) Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs. Mol Gen Genet. 225:289-96. http://www.gmo-compass.org/eng/gmo/db/16.docu.html

12. Super-potato of Fraunhofer Inst. Muth J, Hartje S, Twyman RM, Hofferbert HR, Tacke E, Prüfer D. (2008) Precision breeding for novel starch variants in potato. Plant Biotechnol J. 6:576-84. http://www.fraunhofer.de/en/press/research-news/2009/12/super-potato.jsp “This fall (2009), 100 tonnes of the new super potato that exclusively produces amylopectin were harvested.” Are thesegeneticmodifications or not? Same phenotype, essentially same genotype (loss of function)…

13. Test case 2: herbicideresistance (gain of function) Achievable by different mechanisms/methods: -spontaneous mutation -random mutagenesis -additional gene copies -directed mutation -ketobutyrrate acetolactate ALS CO2 Le et al. (2010) piruvate isoleucine

14. Herbicide tolerant varieties Maize 4,9 Mha in USA - 2002 (15% degli ettari totali a mais) Rapeseed 20% of 4–5 Mha of rapeseed in Canada in 2000 and 2001 Rice 0,74 Mha in USA (2010) Sunflower 0,2 Mha in Turkey (2009) (30% of total surface) Imidazolinoni Sulfonilurea Soybean tolerant to sulfonylurea (STS) cultivated on 3 Mha in 1998 in USA Rice: tolerance achieved through mutagenesis (patent US 5,084,082); reseeding of harvest not allowed by contract HT conventionalvarietiesimply the samesort of risks (mainlyagronomicrisks) astransgenicones, butnobodybothers. Transgenesisallowsstrategies to reduce or mitigate gene flow

15. Varietà CLEARFIELD (frumento) approvata dall’agenzia canadese il 20-03-2003 I. Brief Identification of Plant with Novel Traits (PNT) Designation(s) of the PNT: CLEARFIELD™ wheat line AP602CL Applicant: BASF Canada PlantSpecies: Wheat (Triticumaestivum) Novel Traits: Tolerance to imazamox, an imidazolinoneherbicide Trait Introduction Method: Chemicallyinducedseedmutagenesis Proposed Use: Production of wheat for livestockfeed and human food. Herbicide tolerant variety obtained through mutagenesis! http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0344e.shtml

16. Herbicidetolerantsunflower 150.000 hectarescultivated in 2005 in USA • Cross-breeding towards wild/weedy relatives – YES • Gene flow towardssensitive varieties – YES • Uses more herbicide – YES; imazamox may persist longer in soil. It is often recommended not to grow sensitive varieties within 21 months after herbicide use. Spontaneous resistance is frequent after 4-5 treatments. • Invasive – YES; cultivated sunflower has still some weedy traits. It is often found on roadsides and field borders, but also far away from seed sources. Feral populations can persist in wild environments. Herbicide tolerant variety obtained by classical method and therefore NOT tested for environmental risks. http://www.ars.usda.gov/is/AR/archive/jan05/sunflower0105.htm

17. Conventional approaches often already done or feasible Same character has been achieved by conventional means for: For several other cases, the same character could be ‘easily’ achieved, at least in principle by some conventional method Risks: compared to what?

18. Experimental field: 200m diameter Source: 89 TBq Co-60 in the center Many ‘unnatural’ and coarsemethodswere and are used in creating new varieties -rays used to make novel varieties Institute of Radiation Breeding Ibaraki-ken, JAPAN http://www.irb.affrc.go.jp/

19. Triticale: is it natural? Forced hybridation, embryo culture and chromosome doubling 1874 Scottich botsnist A. S. Wilson crosses wheat and rye. The plant is however sterile 1937 Colchicine treatment achieves chromosome doubling and restores fertility 1970-80 First triticale varieties are released for cultivation 2000 >4M hectares are cultivated nowadays with triticale Isitpossible to foresee the effect of hybridizingwheat and rye and of chromosomedoubling?

20. Consequences of the domestication? What happens when ears of crop cereals fall onto the ground? (non shattering and reduced dormancy) Combined curse:  Germination at the wrong time  Crowding and competition Voluntary maize plantlets in October

21. Voluntary maize plants in mid-November Very high plant density, already damaged by cold  No chances to contribute to the next generation Consequence:  environmental risks mostly irrelevant Non germinating seeds … …rot or are eaten

22. March 27, 1925 340-341 http://depthofprocessing.blogspot.com/2009 /05/are-potato-peels-nutritious.html Poisoning and Toxicology Handbook by Leikin & Paloucek 4th edition, Informa Health Care, 2007 ISBN 1420044796, 9781420044799 Solanine abounds in green parts, sprouts and diseased potatos

23. Put into context http://www.poppyseedtea.com/ Poppy seed tea can kill you

25. Scientificallyspeakingitis a NONSENSE to discuss on the method The only relevant thing is the product: is it dangerous for man? Why? is it dangerous for the environment? Why? Are there benefits? All objections raised against transgenics are applicable also to all plant varieties developed by classical means!

26. Biotechnology and classical breeding work on genes. • Genetic modification of plants by humans takes place since 10.000 years. • No opposition between the methods, no crucial difference Most national and international academies made positive statements on the tecnology Most scientific societies and international organizations (WHO, FAO) concluded on the basis of the published scientific evidence and of accumulated experience after almost 20 years of commercial cultivation, these crops do not present novel or heigher risks compared with the conventional counterparts and provide tangible benefits. for a partial list, see: http://users.unimi.it/morandin/Sources-Academies-societies.doc Academies: USA, Brasil, China, India, Germany, UK, Mexico, France, Italy, Third World, Pontifical Academy of Sciences, Hungary...

27. Implementation? • Some draft laws or regulations remain too long as draft • Some provisions unclear or inconsistent with EU or CPB • Severe restrictions that lack scientific basis • Some fees inhibitory for public research • High burden of information requirements without differentiation • Requests for more information that have no basis in risk assessment • Conflicting views on the interpretations of the laws and regulations between ministries, inspectorates and states • The advice of the scientific bodies is sometimes ignored • Decisions not within legal time frame or not given at all • No adjustment of regulations on the basis of experience • Unworkable conditions, e.g. validated detection in early R&D

28. As of June 1st 2014, 21 safety tested GM crops are being delayed illegally in the EU for a total of 44 years. Additional unjustifiable delays have been increasing since mid-2013, threatening the security of the EU’s food and feed supply.

30. A pathetic case: Italy • Apparently subject to EU regulations • De facto moratorium for commercial cultivation of approved events (Mon 810) • Countless laws and acts in contrast with EU rules. Some legislation bits still missing since 10 years (coexistance) • De facto moratorium of field trials due to missing regulations and conflict between ministries (Agriculture vs. Environment) Short history: http://www.camera.it/leg17/465?area=4&tema=852&OGM+e+utilizzazioni+agrarie#paragrafo4466

31. Production & import (IT) Production / import = 0,47 / 4,07 = 11,5% Source: FAO (http://faostat.fao.org/)

32. Where are theycoming from? Argentina Cake of soybeans

33. Elaborazioni Nomisma (2004) su dati Fao e Eurostat Panelli estratti da soia 8,4% extraUE 15,3% Panelli da import extraUE Panelli estratti da soia italiana o UE Consumo filiere DOP 29,4% Totale consumo 76,3% apparente ~4 M tonn. Grafici di Riccardo Deserti Italy needs transgenic soybeans Italy imports every year (since many years) 4 Mt of soybean & derivatives. Since 2002 imports derive from transgenic soy for more than 50%. Today we are at 70-99%. Must be used even in many DOC and DOP

34. Ifthey are bad, whyimportingthem? Ifthey are good, whynotcultivatingthem? Cosa ne facciamo? Livestocks in Italy (2004-2009)* (Migliaia di capi) *Situation at december 1st of each year except for birds which refers to the whole year Fonte: Assalzoo Without these imports our agro-food sector would collapse

35. Health risks? European corn borer

36. Growth of Fusarium  Fumonisin accumulation

37. Bt maize, field trials

38. Organic maize is less than 1% of the total maize cultivation area but accounts for 31% of the alerts Bt maize is 21% of the total but accounts for 0 alerts

39. Defects ascribable to fumonisins: spina bifida, encefalocele e anencefalia. All in a single summer and a single hospital in Guatemala. Pictures courtesy of Dr. Julio Cabrera. The link between fumonisin, folic acid deficiency and spina bifida quite strong from the biological point of view

40. Quetzaltenango has a mostly indigenous population that consumes high amounts of maize as their staple food   Incidenza dei difetti del tubo neurale (NTD per 10,000 nati vivi) General U.S. popul.: <3 Quetzaltenango: 106 Marasas et al. (2004) U.S.A. Bt maize could have huge benefits in some countries Mean incidence and range in incidence of various locations within the regions or countries are shown; the bar for Limpopo represents one data point. The damages are not only for western consumers. The regulation causes extensive damage to developing countries.

41. Process based regulation – unreasonable Unbearable lightness Estremely costly and thus discriminatory It covers a more precise and predictable technology Obsessioned from risks, agnostic of benefits (72:0) Very damaging (here and abroad!)

42. Sunrise or sunset? • Dawn of green biotechnology passed, but no sunrise in sight for Europe • Excessive regulation, sloppyimplementation and illegal national rules did kill product development and public research in particular • Present regulation favours monopoly • Domino effect on other countries through cultural influence (partic. developing world). Human costs are huge (Golden rice docet)

43. Possibili vie (d’uscita?) • Don’t do anything and hope… • Extend it to classical methods and give breeding a hard time • Revisit regulations (v. difficult: a political decision) • Abide by the rule of law (dossier approval, communication of domestic laws, compliance)

45. Mortalitàglobale 2010 Sorgente: WHO e UN (2010)

46. La carenza di Vitamina A causa Xeroftalmia Principale causa di cecità infantile1 1 Bulletin WHO 2001 79 (3) 214 Vitamina A (Retinolo)

47. www.sightandlife.org Informazionesugliinterventi “traditionali“ per ridurre la VAD si trovano al sito ‘Sightand Life‘ • Distribuzione di pillole di vitamina A. L‘OMS investe 90-100 millioni di $ all‘anno in questoapproccio. • “Educazione“ per unadietadiversificata. • Creazioni di ortidomestici. • Promozione di piantericche in pro-vitA. Nonostanti questi approcci tradizionali, ci sono tuttora 6.000 morti al giorno e 500.000 ciechi ogni anno. Gliinterventitradizionalisonoutilimainsufficienti

48. Fitoene Sintasi dal narciso Fitoene Desaturasi da un batterio

49. Golden rice 2 Narciso Mais Paine, et al., (2005)

50. Quanta provitamina A contiene il Golden Rice e quanto riso dovrebbe mangiare la gente al giorno? Dose giornaliera raccomandata (OMS) in% 140% Riso con 2 g di provitamina A 120% 100% 80% Golden Rice 1 60% 40% Plant Sources 20% Animal source 0% donne bambini • (H Bouis, 2005, unpublished.) Anche con piante basso contenuto di provitamina A (2 g) )una dose normale è sufficiente.