Update on GM wheat field Trials. Huw D Jones Plant Science Dept, Rothamsted Research. UK
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1 Update on GM wheat field Trials Huw D Jones Plant Science Dept, Rothamsted Research. UK
2 Aphids are significant pests of cereals in the UK. Damage grains and also act as vectors for the barley yellow dwarf virus Barley yellow dwarf virus transmitted by cereal aphids
3 When aphids are attacked, either by predators or parasitiods, they produce an alarm pheromone Volatile sesquiterpene: (E)-Beta-farnesene Lacewing larva (Chrysoperla carnea) attacking Aphis fabae
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6 H OPP Farnesyl pyrophosphate (E)-Beta-Farnesene synthase (E)-Beta-Farnesene Also made by some plants
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![Transgenic Arabidopsis expressing CaMV35S:: Eßf synthase cloned from peppermint [mentha x piperita].](/docs-images/90/104150413/images/8-1.jpg "Gas chromatography analysis of volatile chemicals collected from head space of transgenic Arabidopsis and controls wild type transgenic line (E)-β-farnesene Beale, MH, Birkett, MA, Bruce, TJA,")
8 Transgenic Arabidopsis expressing CaMV35S:: Eßf synthase cloned from peppermint [mentha x piperita]. Gas chromatography analysis of volatile chemicals collected from head space of transgenic Arabidopsis and controls wild type transgenic line (E)-β-farnesene Beale, MH, Birkett, MA, Bruce, TJA, Chamberlain, K, Field, LM, Huttly, AK, Martin, JL, Parker, R, Phillips, AL, Pickett, JA, Prosser, IM, Shewry, PR, Smart, LE, Wadhams, LJ, Woodcock, CM & Zhang, Y (2006) Aphid alarm pheromone produced by transgenic plants affects aphid and parasitoid behaviour. PNAS 103:
9 Crop protection tool? Pest Management repel pests attract their enemies
10 A New Generation of Insect resistant GM Crops: Transgenic Wheat synthesising the Aphid Alarm Signal. BBSRC AgriFood FEC 5 yrs PI: John Pickett Researchers: Johnathan Napier, Huw D Jones, Dr Toby Bruce, Lesley Smart, Janet Martin, Angela Doherty
11 Objectives of project: Express Eßf synthase in transgenic wheat and analyse EBF levels Express Eßf synthase + FPP synthase and analyse EBF levels Test whether chloroplast targeting of Eßf synthase + FPP synthase affects overall EBF levels Conduct lab expts. to compare behaviour of aphids / predators / parasitoids on transgenic and control wheat Conduct field trials of two best GM lines
12 Evidence that targeting transgenic terpene synthases to plastids results in enhanced yields of volatile terpenoids Genetic Engineering of Terpenoid Metabolism Attracts Bodyguards to Arabidopsis Kappers et. al. (2005) Science 309: GFP guard cell chloroplasts fluorescence + light image Terpenoid Metabolism in Wild-Type and Transgenic Arabidopsis Plants Aharoni et. al. (2003) Plant Cell 15: We have already demonstrated that it is possible to target GFP to wheat plastids using transit peptides: amyloplasts in wheat endosperm amyloplasts in wheat pollen LF Primavesi, H Wu, EA Mudd, A Day and HD Jones (2008) Visualisation of plastids in endosperm, pollen and roots of transgenic wheat expressing modified GFP fused to transit peptides from wheat SSU RubisCO, rice FtsZ and maize ferredoxin III proteins. Transgenic Research 17(4):
13 EBFS + T EBFS T EBFS + T & FPPS + T EBFS T & FPPS T FPP (Farnesyl pyrophosphate) FPP synthase (targeted or not) H OPP (E)-B-F synthase (targeted or not) EBF (E) B Farnesene
14 Collection of volatiles from transgenic wheat lines A headspace sample of air surrounding the plant is collected on a Porapak Q filter The sample is then analysed for (E) ßfarnesene by GC MS
15 GM Field Trial Application submitted to ACRE /Defra 20 th June 2011
16 4x4 Latin square 6m x 6m plots 2 GM lines plus control Proposed GM wheat field trial design (not draw to scale) 80m 6m 10 m separators (0.5m, 9m barley, 0.5m) 3m 10m 6m 1 GM1 2 Con 3 GM2 4 Con There will be two GM events in the field trial. 10m 5 GM2 6 Con 7 Con 8 GM1 Event 2803R6P1 contains one copy of each of the EBFS+T plasmid and the FPPS+T plasmid per haploid genome and emits pure EBF at high levels 80m 9 Con 10 GM1 11 Con 12 GM2 Event 2812R9P1 contains 4 copies per haploid genome of plasmid EBFS+T only and emits pure EBF at low levels Con GM2 GM1 Con 10 m separators (0.5m, 9m barley, 0.5m) 3m wheat pollen barrier
17 Trial sown on Rothamsted farm for 2 seasons: March - Sept 2012 March - Sept 2013
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19 Other Wheat Field Trials?
20 EC/2001/18 B/GB/11/R8/01 United Kingdom B/DE/10/209 Germany 20/04/2011 B/HU/08/3 Hungary 23/06/2008 B/DE/07/195 Germany 23/06/ /10/2011 Rothamsted Research University of Rostock Department of Biotechnology, Cereal Research University of Rostock Leibniz-Institut of B/DE/06/178 Germany 09/08/2006 Plant Genetics B/IT/04/02 Italy 27/05/2004 Metapontum Agrobios s.c.a r.l. B/ES/04/08 CON Spain 05/02/2004 Instituto de Agricol. Sostenibile Consejo Study of aphid, predator and parasitoid behaviour in wheat producing aphid alarm pheromone Evaluation of the resistance of KP4 bearing GM spring wheat plants against loose smut in the field with special consideration of effects on resistance against other fungal pathogens Field testing of genetically modified wheat lines for improved red rust resistance Field evaluation of loose smut resistance in GM spring wheat expressing KP4 protein with focus on resistance against other fungal pathogens. Increase of grain protein content in winter wheat Study of the stability of the transgene and his hereditability of genetically modified wheat under of open field conditions. Evaluation in field conditions of Fungal resistant wheat. B/DE/03/151 Germany 26/01/2004 Syngenta Seeds Fungal resistant wheat Germany 2004 (I) & (II) B/DE/02/143 Germany 19/05/2003 Syngenta Seeds Fungal resistant wheat Germany Regs B/ES/02/17 Spain 06/02/2003 B/02/R34/4 02/R36/1 01/R8/4 97/R8/3 United Kingdom United Kingdom United Kingdom United Kingdom Instituto Nacional de Investigaciones INIA 16/01/2003 Syngenta Seeds Ltd 16/04/2002 Long Ashton RS Rothamsted 16/04/2002 Research Rothamsted 06/01/1998 Research Safety assessment of the release of transgenic crops spread of herbicide resistance genes from wheat. To compare the pathogen infestation level and mycotoxin level of wheat expressing an enhanced resistance to Fusarium pathogens with existing non-modified varieties, grown under standard agronomic conditions. Small scale trial of spring wheat to assess control of sucrose synthase activity and starch accumulation in the endosperm. Improved bread making performance of commercial UK wheat Improved bread making performance of spring wheat
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23 Acknowledgements GM Wheat Angela Doherty Caroline Sparks Johnathan Napier Chemical Ecology Toby Bruce Lesley Smart Gudbjorg Aradottir Janet Martin John Pickett Rothamsted Research receives grant aided support from the BBSRC. This project was supported by BBSRC grant BBG Field Trial Stephen Goward
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