Improving the nitrogen responses of UK wheat varieties
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Dr. Yongfang Wan, Dr. Malcolm Hawkesford, Dr. Rowan Mitchell, Dr. Peter Shewry Rothamsted Research, UK
Improving the nitrogen responses of UK wheat varieties
- 1. Improving the nitrogen responses of UK wheat varieties Yongfang Wan, Malcolm Hawkesford, Rowan Mitchell and Peter Shewry Rothamsted Research, UK
- 2. Background • Most varieties exhibit a negative relationship between grain protein and yield. • Few varieties show reproducible deviations, such as Cordiale and Marksman with high yield and grain protein.
- 3. • These varieties can make most efficient use of nitrogen inputs, have better economic costs and minimise environmental pollution. • Little is known about the molecular basis for the protein deviation resulting from: • nitrogen uptake • source metabolism • sink activity
- 4. This project will: • identify some candidate transcripts responding to nitrogen application and /or related to the protein deviation; • determine the effect of nitrogen levels on protein composition; • examine interaction between nitrogen, protein deviation and bread-making quality.
- 5. Strategy Field trials Three N levels Six cultivars (N100, N200, (He, Co, Mk, Is, N350kg/ha) Ma, Xi) AA 14 DAA AA 21DAA AA AA AA 28 DAA 35DAA 42DAA mature Protein Baking Transcriptome Agronomic traits Affymetrix and qPCR composition quality Grain yield and protein SE-HPLC;SDS-PAGE integrated Bioinformatics Hypothesis, Genes, markers
- 6. Grain protein responses to nitrogen in 2009 and 2010 at Rothamsted. 3 2.5 2 Grain %N N100 1.5 N200 N350 1 0.5 0 Co He Is Ma Mk Xi Co He Is Ma Mk Xi 2009 I 2010
- 7. Grain yield responses to nitrogen in 2009 and 2010 at Rothamsted14 12 10 Grain yield (t/ha@85% DM) 8 N100 N200 6 N350 4 2 0 Co He Is Ma Mk Xi Co He Is Ma Mk Xi 2009 I 2010
- 8. Transcriptome analysis at 21D in 2009 and 2010 by Affymetrix . Storage 2009 2010 proteins(13) Defence genes (6) photosynthesis genes(5) Unknown N100 N200 N350 N100 N200 N350 genes (81) 105 transcripts are significantly up-regulated by nitrogen.
- 9. Response of storage protein transcripts to nitrogen Gliadin transcripts 15000 14000 Raw expression 13000 12000 11000 2009 10000 2010 N100 N200 N350 N100 N200 N350 N100 N200 N350 N100 N200 N350 N100 N200 N350 N100 N200 Co N350 He Is LMW-glutenin subunit transcripts Ma Mk Xi 30000 Raw expression HMW-glutenin subunit transcripts 25000 20000 22000 15000 2009 20000 10000 raw expression 2010 N100 N350 N200 N100 18000 2009 N350 N200 N100 N350 N200 16000 Co He 14000 Is Ma Mk Xi 2009 12000 2010 10000 N100 N200 N350 N100 N200 N350 N100 N200 N350 N100 N200 2009 N350 N100 N200 N350 N100 N200 N350 Co He Is Ma Mk Xi
- 10. A novel γ-gliadin gene is up-regulated by nitrogen. Expression on Affymetrix 14000 2009 14000 2010 12000 12000 Expression intensity 10000 10000 8000 8000 6000 6000 N100 N100 4000 4000 N200 N200 2000 2000 N350 N350 0 0 CO HE IS MA MK XI CO HE IS MA MK XI 10 9 8 RELATIVE EXPRESSION 7 N100 6 5 N200 4 N350 3 2 1 0 14 21 28 35 days after anthesis Expression by realtime RT-PCR
- 11. Comparison of novel γ-gliadin genes CS-typical γ-gliadin C γ-3 hordein (Hordeum chilense) S CS-3 - T. monococcum 1AS 3 Ae. tauchii CS-1 1DS He-1 CS-2 1BS He-2
- 12. Effect of nitrogen on protein composition Xi IS Co Ma MK He N100 N200 N350 Ax Dx Bx HMW-glutenins By Dy ω- gliadins LMW-glutenins & α,β,γ gliadins Globulin and Albumin
- 13. ω- 5 ω- (1+2) By Dx Dy Bx HMW glutenins ω - gliadins LMW-glutenin & gliadins Dx2%=Dx2 volume/total HMW glutenins
- 14. Cordiale 32.5 2009 2010 32.5 27.5 27.5 22.5 N100 22.5 N100 HMW-glutenin 17.5 N200 17.5 N200 N350 N350 12.5 12.5 7.5 7.5 1Ax1 1Dx5 1Bx17 1By18 1Dy10 1Ax1 1Dx5 1Bx17 1By18 1Dy10 70.0 70.0 60.0 60.0 50.0 50.0 N100 40.0 N100 40.0 N200 Omega-gliadin 30.0 N350 30.0 N200 20.0 20.0 N350 10.0 10.0 0.0 0.0 ω-5a ω-(1+2)a ω-(1+2)b ω-5a ω-(1+2)a ω-(1+2)b 80.0 90.00 70.0 80.00 70.00 60.0 60.00 50.0 N100 50.00 N100 40.0 40.00 N200 Group protein 30.0 N200 N350 30.00 N350 20.00 20.0 10.00 10.0 0.00 HMW-GS% ω-GLIADIN% LMW-GS+GLIADINS (%) 0.0 HMW-GS% ω-GLIADIN% LMW-GS+GLIADINS (%)
- 16. Relationship of polymeric and monomeric proteins by SE-HPLC in 2009 mature seeds 40.0 HE 39.0 HE HE 38.0 Polymeric (F1+F2) 37.0 Ma Linear (N100) 36.0 CO MK MK Linear (N200) 35.0 Linear (N350) 34.0 XI IS IS IS IS 33.0 IS 32.0 N100 N200 N350 40.0 42.0 44.0 46.0 48.0 50.0 52.0 Monomeric (F3+F4)
- 17. 2010 baking quality from Rothamsted site
- 18. Summary 1: Nitrogen application can increase both grain protein and yield. 2: The storage protein transcripts are significantly up-regulated by nitrogen. 3: The protein compositions respond to nitrogen disproportionally. 4: The baking quality is improved by higher nitrogen levels, but is affected by different sites, and years .
- 19. Future Nitrogen environments Protein Grain protein Gene expression composition Grain yield Protein deviation Baking quality
- 20. Acknowledgment Funding: Partners: Campden BRI, Gemma Chope HGCA, Dhan Bhandari Nickerson Seeds RAGT seeds Syngenta seeds KWS-UK ADM milling Ltd Heygates ltd Warbourtons Rank Hovis McDougal