Insight into the genetic basis of winter hardiness and the potential this has to alter US malting-quality barley

Transcription

1 Eric J Stockinger AMBA Barley Improvement Conference San Diego, CA January 14, 2013 Insight into the genetic basis of winter hardiness and the potential this has to alter US malting-quality barley

2 Winter hardiness The plant breeder who developed Maris Otter and Director of the Plant Breeding Institute in Cambridge England, Dr. George Douglas Hutton Bell (G.D.H. Bell), 1944: there is no absolute difference between spring and winter types, and between hardy and non-hardy types, but the differences are essentially quantitative showing a gradation from extreme early spring types to extreme winter types, and from lack of hardiness to extreme winter hardiness. Bell GDH, The breeding of two-row winter-hardy barley. Journal of Agricultural Science 34,

3 Winter hardiness Complete winter kill (2002/2003) Partial winter kill (2006/2007)

4 Winter vs. spring types Winter types Plants in vegetative phase Vernalization induces reproductive phase Spring types Inherent reproductive competence

5 Barley types cultivated in different regions in 1952 Poehlman JM (1952) Breeding winter barley for hardiness and disease resistance. Econ. Bot. 6,

6 Screening winter barley germplasm (Columbia MO, winter ) Number of varieties with survival of Variety source 0% 1 20% 21 40% 41 60% 61 80% % Tennessee Winter Korea China-Manchuria Caucasus Western Europe Mediterranean Ethiopia Japan India Poehlman JM (1952) Breeding winter barley for hardiness and disease resistance. Econ. Bot. 6,

7 The top 10 winter-hardy barleys (of 204 assayed ) 1. Kearney 2. Dicktoo 3. OAC 4GH1 4. Dayton 5. Nebraska Nebraska Admire 8. Kansas 9. OAC 30GH PU B466A7772 Wiebe GA and Reid DA (1958) Comparative winter hardiness of barley varieties. Technical Bulletin. United States Department of Agriculture 1176, 20 pp.

8 Pedigree of the most successful winter malting barley Fischbeck G (1992) Barley cultivar development in Europe - success in the past and possible changes in the future. In Barley Genetics VI; Proceedings of the Sixth International Barley Genetics Symposium. Munck L (ed.), (ed.), Vol. II, pp , Munksgaard International Publishers, Ltd, Copenhagen, Denmark, Helsingborg, Sweden.

9 Potential for winter malting barley in the US Variety Row On 6/64" Winter survival (%) *, (%) * Tschermak 2 46% 12% Carstens 2 52% 23% MO B % 57% * Based on three Missouri locations Minimum acceptable: 6-row 70%; 2-row 80% Duelos LA, Poehlman JM, and Hoskins PH (1970) Breeding 2-row winter-type malting barley. In Barley genetics II. Proceedings of Second International Barley Genetics Symposium. (ed.), Vol. pp , Washington State University, Pullman.

10 Winter survival of lines selected (Columbia Missouri 1969) Cross or accession Percentage of F 6 lines in percent survival class 1 19% 20 39% 40 59% 60 79% 80 99% MO B475 Carstens Ludwig Carstens MO B1300 Carstens MO B1400 Carstens MO B475 Tschermak Ludwig Tschermak MO B1337 Tschermak MO B1400 Tschermak MO B Carstens Duelos LA, Poehlman JM, and Hoskins PH (1970) Breeding 2-row winter-type malting barley. In Barley genetics II. Proceedings of Second International Barley Genetics Symposium. (ed.), Vol. pp , Washington State University, Pullman.

11 Winter survival (Columbia Missouri, ) Selection Pedigree Survival (%) MO B2126 MO B475 Tschermak 77 MO B2185 MO B1400 Tschermak 76 MO B2186 MO B1400 Tschermak 77 MO B2190 MO B1400 Tschermak 69 MO B2191 MO B1400 Tschermak 66 MO B2192 MO B1400 Tschermak 72 MO B475 Admire Missouri Early Beardless 81 Carstens 58 Poehlman JM, Duclos L, Kruse C. (1973) Progress in development of two-row winter malting barley. Proceedings of the Barley Improvement Conference, Minneapolis, Minnesota,

12 Genetic analyses of winter hardiness (the Dicktoo Morex population) Dicktoo LT 50 = -8 o C (Six-row winter) Morex LT 50 = -4 o C (Six-row spring) F 1 Generate 100 doubled haploid lines from F 1 creating an immortalized F 2 population Winter hardiness: Score field survival in Oregon and Montana Freezing tolerance: Score regrowth after controlled freezes at 0, -4, -8, -12 C Associated biochemical traits: Score field material for crown fructan content Quantitative trait loci analysis (QTL): Test for association between traits and regions of the barley genome Hayes PM, Blake T, Chen THH, et al., Quantitative trait loci on barley (Hordeum vulgare L.) chromosome 7 associated with components of winterhardiness. Genome 36,

13 Genetic analyses of winter hardiness (the Nure Tremois population) Nure LT 50 = -11 o C (Two-row winter) Tremois LT 50 = -6 o C (Two-row spring) F 1 Generate 136 doubled haploid lines from F 1 creating an immortalized F 2 population Winter hardiness: Score field survival in Northern Italy Freezing tolerance: Score regrowth after controlled freezes at -11 and -13 o C Associated biochemical traits: Score field material COR14B protein accumulation Quantitative trait loci analysis (QTL): Test for association between traits and regions of the genome Francia E, Rizza F, Cattivelli L, et al., Two loci on chromosome 5H determine low-temperature tolerance in a 'Nure' (winter) 'Tremois' (spring) barley map. Theor. Appl. Genet. 108,

14 Two loci have major effects on winter hardiness CBF VRN-H1 Francia E, Rizza F, Cattivelli L, et al., Two loci on chromosome 5H determine low-temperature tolerance in a 'Nure' (winter) 'Tremois' (spring) barley map. Theor. Appl. Genet. 108, Skinner JS, Szucs P, Von Zitzewitz J, et al., Mapping of barley homologs to genes that regulate low temperature tolerance in Arabidopsis. Theor. Appl. Genet. 112,

15 CBFs binds CRT/DREs and activate genetic pathways increasing freezing tolerance Stockinger et. al., 1997; Gilmour et. al., 1998; Jaglo-Ottosen et. al., 1998

16 FROST RESISTANCE-H2 FR-H2 has >13 CBFs In Dicktoo and Nure, CBF2A and CBF4B are present in multiple copies in tandem array Nure Tremois In Morex and Tremois, CBF2 and CBF4 are present in single copy Knox AK, Dhillon T, Cheng H, Tondelli A, Pecchioni N, Stockinger EJ, CBF gene copy number variation at Frost Resistance-2 is associated with levels of freezing tolerance in temperate-climate cereals. Theor. Appl. Genet. 121, Francia E, Barabaschi D, Tondelli A, et al., Fine mapping of a HvCBF gene cluster at the frost resistance locus Fr-H2 in barley. Theor. Appl. Genet. 115,

17 CBF14 copy numbers vary across wheats (and between classes) Hard Red Wheats (Crimean landrace origins) Soft Red Wheats (W. N. European landrace origins) B A D Knox AK, Dhillon T, Cheng H, Tondelli A, Pecchioni N, Stockinger EJ, CBF gene copy number variation at Frost Resistance-2 is associated with levels of freezing tolerance in temperate-climate cereals. Theor. Appl. Genet. 121,

18 Rhg1 copy number variation in soybean Cook DE, Lee TG, Guo X, et al., Copy number variation of multiple genes at Rhg1 mediates nematode resistance in soybean. Science 338,

19 2012 Malting quality data Accession On 6/64 (%) Barley Protein (%) DP ( o ASBC) Alphaamylase (20 o DU) Beta-glucan (ppm) Quality Score Overall Rank 'Maja' 'Endeavor' * 'Charles * 'Wintmalt' 'Maris Otter' 'Halcyon' 'Pipkin' 'Puffin' 'Signal' 'Trigger' MO B MO B MO B MO B MO B MO B * Blue bold text = values below ideal minimum AMBA quality standard Green bold text = values above ideal maximum AMBA quantity standard

20 Winter survival and freezing tolerance Accession Winter survival (Elora Research Station) Fv/Fm Control -6 o C -8 o C -10 o C -12 o C Cassata Flagon Maris Otter Pearl Macgregor McKellar H H H H H H Kawartha (SB) n.a Maxine (WW) n.a Larson, R. J. A. (2012) Winter hardy spring wheat breeding: analysis of winter spring wheat germplasm and the development of selection tools. Ph.D. Thesis, University of Guelph.

21 What is the underlying molecular basis of FR-H1?

22 Insight into the relationship between VRN-1 and freezing tolerance (FR-1) Vrn-1 spring allele: Plants in reproductive phase VRN-1 transcripts accumulate Vernalization not required vrn-1 winter allele: Plants in vegetative phase VRN-1 transcript accumulation restricted Vernalization induces VRN-1 expression

23 CBFs and VRN-1 are inversely expressed and are differentially affected by vernalization Nonvernalized: CBFs on VRN-1 off Vernalized: CBFs off VRN-1 on Stockinger EJ, Skinner JS, Gardner KG, Francia E, Pecchioni N, Expression levels of barley Cbf genes at the Frost resistance- H2 locus are dependent upon alleles at Fr-H1 and Fr-H2. Plant J. 51,

24 CBFs and VRN-1 are inversely expressed and are differentially affected by vernalization Nonvernalized: CBFs on VRN-1 off Vernalized: CBFs off VRN-1 on Stockinger EJ, Skinner JS, Gardner KG, Francia E, Pecchioni N, Expression levels of barley Cbf genes at the Frost resistance- H2 locus are dependent upon alleles at Fr-H1 and Fr-H2. Plant J. 51,

25 'Dicktoo' Short days: CBFs on VRN-1 off Long days: CBFs off VRN-1 on CBF2 CBF4 CBF9 PHYC ADA2 VRN-H1 Stockinger EJ, Skinner JS, Gardner KG, Francia E, Pecchioni N, Expression levels of barley Cbf genes at the Frost resistance- H2 locus are dependent upon alleles at Fr-H1 and Fr-H2. Plant J. 51,

26 Key findings and prospects Regulatory genes increasing freezing tolerance, the CBFs, are higher in copy number in winter barley lines than spring barley lines. FR-H2 in part may be a numbers game in which more is better High copy number Low copy number

27 Key findings and prospects Activity (expression) of CBF genes at FR-2 shows inverse relationship with VRN-H1 VRN-H1 may be a negative regulator of the CBFs

28 Regional Barley Gene Mapping Project