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Grain dormancy loss is associated with changes in ABA and GA sensitivity and hormone accumulation in bread wheat, Triticum aestivum (L.)

Published online by Cambridge University Press:  09 March 2015

Keiko M. Tuttle ,
Shantel A. Martinez ,
Elizabeth C. Schramm ,
Yumiko Takebayashi ,
Mitsunori Seo  and
Camille M. Steber
Keiko M. Tuttle
Affiliation:
Molecular Plant Sciences, Department of Crop and Soil Sciences, Washington State University, Pullman, WA99164-6420, USA
Shantel A. Martinez
Affiliation:
Molecular Plant Sciences, Department of Crop and Soil Sciences, Washington State University, Pullman, WA99164-6420, USA
Elizabeth C. Schramm
Affiliation:
Molecular Plant Sciences, Department of Crop and Soil Sciences, Washington State University, Pullman, WA99164-6420, USA
Yumiko Takebayashi
Affiliation:
RIKEN Centre for Sustainable Resource Science, Yokohama, Kanagawa230-0045, Japan
Mitsunori Seo
Affiliation:
RIKEN Centre for Sustainable Resource Science, Yokohama, Kanagawa230-0045, Japan
Camille M. Steber*
Affiliation:
Molecular Plant Sciences, Department of Crop and Soil Sciences, Washington State University, Pullman, WA99164-6420, USA USDA-ARS Wheat Genetics, Quality, Physiology and Disease Research Unit, Washington State University, Pullman, WA99164-6420, USA
*
*Correspondence E-mail: [email protected]
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Abstract

Knowledge about the hormonal control of grain dormancy and dormancy loss is essential in wheat, because low grain dormancy at maturity is associated with the problem of pre-harvest sprouting (PHS) when cool and rainy conditions occur before harvest. Low GA (gibberellin A) hormone sensitivity and high ABA (abscisic acid) sensitivity were associated with higher wheat grain dormancy and PHS tolerance. Grains of two PHS-tolerant cultivars were very dormant at maturity, and insensitive to GA stimulation of germination. More PHS-susceptible cultivars were less sensitive to ABA inhibition of germination, and were either more GA sensitive or germinated efficiently without GA at maturity. As grain dormancy was lost through dry afterripening or cold imbibition, grains first gained GA sensitivity and then lost ABA sensitivity. These changes in GA and ABA sensitivity can serve as landmarks defining stages of dormancy loss that cannot be discerned without hormone treatment. These dormancy stages can be used to compare different cultivars, seed lots and studies. Previous work showed that wheat afterripening is associated with decreasing ABA levels in imbibing seeds. Wheat grain dormancy loss through cold imbibition also led to decreased endogenous ABA levels, suggesting that reduced ABA signalling is a general mechanism triggering dormancy loss.

Keywords

abscisic acidafterripeningcold stratificationdormancygibberellinpre-harvest sproutingTriticum aestivum
Type
Research Papers
Information
Seed Science Research , Volume 25 , Special Issue 2: Seed Dormancy , June 2015 , pp. 179 - 193
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Copyright
Copyright © Cambridge University Press 2015. This is a work of the U.S. Government and is not subject to copyright protection in the United States. 

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References

Anderson, J.A., Sorrells, M.E. and Tanksley, S.D. (1993) RFLP analysis of genomic regions associated with resistance to preharvest sprouting in wheat. Crop Science 33, 453–459.CrossRefGoogle Scholar
Appleford, N.E.J., Evans, D.J., Lenton, J.R., Gaskin, P., Croker, S.J., Devos, K.M., Phillips, L.A. and Hedden, P. (2006) Function and transcript analysis of gibberellin-biosynthetic enzymes in wheat. Planta 223, 568–582.CrossRefGoogle ScholarPubMed
Appleford, N.E.J., Wilkinson, M.D., Ma, Q., Evans, D.J., Stone, M.C., Pearce, S.P., Powers, S.J., Thomas, S.G., Jones, H.D., Phillips, A.L., Hedden, P. and Lenton, J.R. (2007) Decreased shoot stature and grain α-amylase activity following ectopic expression of a gibberellin 2-oxidase gene in transgenic wheat. Journal of Experimental Botany 58, 3213–3226.CrossRefGoogle ScholarPubMed
Ariizumi, T., Hauvermale, A.L., Nelson, S.K., Hanada, A., Yamaguchi, S. and Steber, C.M. (2013) Lifting DELLA repression of Arabidopsis seed germination by nonproteolytic gibberellin signalling. Plant Physiology 162, 2125–2139.CrossRefGoogle Scholar
Barrero, J.M., Talbot, M.J., White, R.G., Jacobsen, J.V. and Gubler, F. (2009) Anatomical and transcriptomic studies of the coleorhiza reveal the importance of this tissue in regulating dormancy in barley. Plant Physiology 150, 1006–1021.CrossRefGoogle ScholarPubMed
Barrero, J.M., Millar, A.A., Griffiths, J., Czechowski, T., Scheible, W.R., Udvardi, M., Reid, J.B., Ross, J.J., Jacobsen, J.V. and Gubler, F. (2010) Gene expression profiling identifies two regulatory genes controlling dormancy and ABA sensitivity in Arabidopsis seeds. The Plant Journal 61, 611–622.CrossRefGoogle ScholarPubMed
Baskin, J.M. and Baskin, C.C. (2004) A classification system for seed dormancy. Seed Science Research 14, 1–16.CrossRefGoogle Scholar
Benech-Arnold, R.L., Giallorenzi, C.M., Frank, J. and Rodriguez, V. (1999) Termination of hull-imposed dormancy in developing barley grains is correlated with changes in embryonic ABA levels and sensitivity. Seed Science Research 9, 39–47.CrossRefGoogle Scholar
Bewley, D.J. and Black, M. (1994) Seeds: Physiology of development and germination. New York, Plenum Press.CrossRefGoogle Scholar
Bewley, J.D., Bradford, K.J., Hilhorst, H.W.M. and Nonogaki, H. (2013) Seeds: Physiology of development, germination and dormancy (3rd edition). New York, Springer.CrossRefGoogle Scholar
Campbell, K.G., Allan, R.E., Anderson, J., Burke, A., Blake, N., Hoagland, C., Walker, C., Chatelain, J., Little, L.M., Pritchett, J., Chen, X., Morris, C., See, D., Guy, S., Murray, T., Engle, D., Wetzel, H. and Wood, D. (2013) Registration of ‘Cara’ soft white winter club wheat. Journal of Plant Registrations 7, 81–88.CrossRefGoogle Scholar
Carter, A.H., Jones, S.S., Cai, X., Lyon, S.R., Balow, K.A., Shelton, G.B., Higginbotham, R.W., Chen, X.W., Engle, D.A., Baik, B., Guy, S.O., Murray, T.D. and Morris, C.F. (2014) Registration of ‘Puma’ soft white winter wheat. Journal of Plant Registrations 8, 273–278.CrossRefGoogle Scholar
Cereal Variety Testing . (2013) Western Washington variety trials. Available from http://variety.wsu.edu/ and http://plantbreeding.wsu.edu/varietyTrialsWesternWA.html (accessed 20 November 2014).Google Scholar
Debeaujon, I., Lepiniec, L., Pourcel, L. and Routaboul, J.M. (2007) Seed coat development and dormancy. pp. 25–49 in Bradford, K.J.; Nonogaki, H. (Eds) Seed development, dormancy and germination. Annual Plant Reviews Vol. 27. Sheffield, UK, Blackwell Publishing.CrossRefGoogle Scholar
DePauw, R.M., Knox, R.E., Singh, A.K., Fox, S.L., Humphreys, D.G. and Hucl, P. (2012) Developing standardized methods for breeding preharvest sprouting resistant wheat, challenges and successes in Canadian wheat. Euphytica 188, 7–14.CrossRefGoogle Scholar
Derkx, M.P.M. and Karssen, C.M. (1993) Effects of light and temperature on seed dormancy and gibberellin-stimulated germination in Arabidopsis thaliana: studies with gibberellin-deficient and -insensitive mutants. Physiologia Plantarum 89, 360–368.CrossRefGoogle Scholar
Fang, J., Chai, C., Qian, Q., Li, C., Tang, J., Sun, L., Huang, Z., Guo, X., Sun, C., Liu, M., Zhang, Y., Lu, Q., Wang, Y., Lu, C., Han, B., Chen, F., Cheng, Z. and Chu, C. (2008) Mutations of genes in synthesis of the carotenoid precursors of ABA lead to pre-harvest sprouting and photo-oxidation in rice. Plant Journal 54, 177–189.CrossRefGoogle ScholarPubMed
Finkelstein, R.R., Reeves, W., Ariizumi, T. and Steber, C.M. (2008) Molecular aspects of seed dormancy. Annual Review of Plant Biology 59, 387–415.CrossRefGoogle ScholarPubMed
Flintham, J.E. (2000) Different genetic components control coat-imposed and embryo-imposed dormancy in wheat. Seed Science Research 10, 43–50.CrossRefGoogle Scholar
Gao, F., Jordan, M.C. and Ayele, B.T. (2012) Transcriptional programs regulating seed dormancy and its release by after-ripening in common wheat (Triticum aestivum L.). Plant Biotechnology Journal 10, 465–476.CrossRefGoogle ScholarPubMed
Gerjets, T., Scholefield, D., Foulkes, J.M., Lenton, J.R. and Holdsworth, M.J. (2010) An analysis of dormancy, ABA responsiveness, after-ripening and pre-harvest sprouting in hexaploid wheat (Triticum aestivum L.) caryopses. Journal of Experimental Botany 61, 597–607.CrossRefGoogle ScholarPubMed
Hedden, P. and Kamiya, Y. (1997) Gibberellin biosynthesis: enzymes, genes and their regulation. Annual Review of Plant Physiology and Plant Molecular Biology 48, 431–460.CrossRefGoogle ScholarPubMed
Heyne, E.G. (1959) Registration of improved wheat varieties, XXIII. Agronomy Journal 51, 689–692.CrossRefGoogle Scholar
Himi, E., Mares, D.J., Yanagisawa, A. and Noda, K. (2002) Effect of grain colour gene (R) on grain dormancy and sensitivity of the embryo to abscisic acid (ABA) in wheat. Journal of Experimental Botany 53, 1569–1574.CrossRefGoogle ScholarPubMed
Himi, E., Maekawa, M., Miura, H. and Noda, K. (2011) Development of PCR markers for Tamyb10 related to R-1, red grain color gene in wheat. Theoretical and Applied Genetics 122, 1561–1576.CrossRefGoogle ScholarPubMed
Hori, K., Sugimoto, K., Nonoue, Y., Ono, N., Matsubara, K., Yamanouchi, U., Abe, A., Takeuchi, Y. and Yano, M. (2010) Detection of quantitative trait loci controlling pre-harvest sprouting resistance by using backcrossed populations of japonica rice cultivars. Theoretical and Applied Genetics 120, 1547–1557.CrossRefGoogle ScholarPubMed
Jacobsen, J.V. and Chandler, P.M. (1987) Gibberellin and abscisic acid in germinating cereals. pp. 164–193 in Davies, P.J. (Ed.) Plant hormones and their role in plant growth and development. New York, Springer Publishing.CrossRefGoogle Scholar
Jacobsen, J.V., Pearce, D.W., Poole, A.T., Pharis, R.P. and Mander, L.N. (2002) Abscisic acid, phaseic acid and gibberellin contents associated with dormancy and germination in barley. Physiologia Plantarum 115, 428–441.CrossRefGoogle ScholarPubMed
Jacobsen, J.V., Barrero, J.M., Hughes, T., Julkowska, M., Taylor, J.M., Xu, Q. and Gubler, F. (2013) Roles for blue light, jasmonate and nitric oxide in the regulation of dormancy and germination in wheat grain (Triticum aestivum L.). Planta 238, 121–138.CrossRefGoogle ScholarPubMed
Jones, S.S., Murray, T.D., Lyon, S.R., Morris, C.F. and Line, R.F. (2001) Registration of ‘Bruehl’ wheat. Crop Science 41, 2006–2007.CrossRefGoogle Scholar
Jones, S.S., Lyon, S.R., Balow, K.A., Gollnick, M.A., Murphy, K.M., Kuehner, J.S., Murray, T.D., Chen, X.M., Engle, D.A. and Campbell, K.G. (2010) Registration of ‘Xerpha’ wheat. Journal of Plant Registrations 4, 137–140.CrossRefGoogle Scholar
Karssen, C. and Lacka, E. (1986) A revision of the hormone balance theory of seed dormancy: studies on gibberellin and/or abscisic acid-deficient mutants of Arabidopsis thaliana . pp. 315–323 in Bopp, M. (Ed.) Plant growth substances 1985: Proceedings of the 12th International Conference on Plant Growth Substances. New York, Springer-Verlag.CrossRefGoogle Scholar
Karssen, C.M., der Brinkhorst-van Swan, D.L.C., Breekland, A.E. and Koornneef, M. (1983) Induction of dormancy during seed development by endogenous abscisic acid: studies in abscisic acid deficient genotypes of Arabidopsis thaliana (L.) Heynh. Planta 157, 158–165.CrossRefGoogle ScholarPubMed
Kawakami, N., Miyake, Y. and Noda, K. (1997) ABA insensitivity and low ABA levels during seed development of non-dormant wheat mutants. Journal of Experimental Botany 48, 1415–1421.CrossRefGoogle Scholar
King, R.W. and Richards, R.A. (1984) Water uptake in relation to pre-harvest sprouting damage in wheat: ear characteristics. Australian Journal of Agricultural Research 35, 327–336.CrossRefGoogle Scholar
King, R.W. and von Wettstein-Knowles, P. (2000) Epicuticular waxes and regulation of ear wetting and pre-harvest sprouting in barley and wheat. Euphytica 112, 157–166.CrossRefGoogle Scholar
Kobayashi, F., Takumi, S. and Nakamura, C. (2008) Increased freezing tolerance in an ABA-hypersensitive mutant of common wheat. Journal of Plant Physiology 165, 224–232.CrossRefGoogle Scholar
Kucera, B., Cohn, M.A. and Luebner-Metzger, G. (2005) Plant hormone interactions during seed dormancy release and germination. Seed Science Research 15, 281–307.CrossRefGoogle Scholar
Kushiro, T., Okamoto, M., Nakabayashi, K., Yamagishi, K., Kitamura, S., Asami, T., Hirai, N., Koshiba, T., Kamiya, Y. and Nambara, E. (2004) The Arabidopsis cytochrome P450 CYP707A encodes ABA 8′-hydroxylases: key enzymes in ABA catabolism. EMBO Journal 23, 1647–1656.CrossRefGoogle Scholar
Li, C., Ni, P., Francki, M., Hunter, A., Zhang, Y., Schibeci, D., Li, H., Tarr, A., Wang, J., Cakir, M., Yu, J., Bellgard, M., Lance, R. and Appels, R. (2004) Genes controlling seed dormancy and pre-harvest sprouting in a rice–wheat–barley comparison. Functional and Integrative Genomics 4, 84–93.CrossRefGoogle Scholar
Liu, A., Gao, F., Kanno, Y., Jordan, M.C., Kamiya, Y., Seo, M. and Ayele, B.T. (2013) Regulation of wheat seed dormancy by after-ripening is mediated by specific transcriptional switches that induce changes in seed hormone metabolism and signalling. PloS ONE 8, e56570.CrossRefGoogle Scholar
McCaig, T. and DePauw, R. (1992) Breeding for preharvest sprouting tolerance in white-seed-coat spring wheat. Crop Science 32, 19–23.CrossRefGoogle Scholar
McMaster, G.J. and Derera, N.F. (1976) Methodology and sample preparation when screening for sprouting damage in cereals. Cereal Research Communications 4, 251–254.Google Scholar
Millar, A.A., Jacobsen, J.V., Ross, J.J., Helliwell, C.A., Poole, A.T., Scofield, G., Reid, J.B. and Gubler, F. (2006) Seed dormancy and ABA metabolism in Arabidopsis and barley: The role of ABA 8′-hydroxylase. Plant Journal 45, 942–954.CrossRefGoogle ScholarPubMed
Miyamoto, T. and Everson, E.H. (1958) Biochemical and physiological studies of wheat seed pigmentation. Agronomy Journal 50, 733–734.CrossRefGoogle Scholar
Morris, C.F. and Paulsen, G.M. (1989) Registration of five preharvest sprouting-resistant hard white winter wheat germplasm lines. Crop Science 29, 246–247.CrossRefGoogle Scholar
Morris, C., Moffatt, J.M., Sears, R.G. and Paulsen, G.M. (1989) Seed dormancy and responses of caryopses, embryos, and calli to abscisic acid in wheat. Plant Physiology 90, 643–647.CrossRefGoogle ScholarPubMed
Morris, C., Anderberg, R.J., Goldmark, P.J. and Walker-Simmons, M. (1991) Molecular cloning and expression of abscisic acid-responsive genes in embryos of dormant wheat seeds. Plant Physiology 95, 814–821.CrossRefGoogle ScholarPubMed
Nakamura, S., Abe, F., Kawahigashi, H., Nakazono, K., Tagiri, A., Matsumoto, T., Utsugi, S., Ogawa, T., Handa, H., Ishida, H., Mori, M., Kawaura, K., Ogihara, Y. and Miura, H. (2011) A wheat homolog of MOTHER OF FT and TFL1 acts in the regulation of germination. Plant Cell 23, 3215–3229.CrossRefGoogle ScholarPubMed
Noda, K., Kawabata, C. and Kawakami, N. (1994) Response of wheat grain to ABA and imbibition at low temperature. Plant Breeding 113, 53–57.CrossRefGoogle Scholar
Nyachiro, J.M., Clarke, F.R., DePauw, R.M., Knox, R.E. and Armstrong, K.C. (2002) Temperature effects of seed germination and expression of seed dormancy in wheat. Euphytica 126, 123–127.CrossRefGoogle Scholar
Paterson, A., Sorrells, M. and Obendorf, R. (1989) Methods of evaluation for preharvest sprouting resistance in wheat breeding programs. Canadian Journal of Plant Science 69, 681–689.CrossRefGoogle Scholar
Pope, W. (1978) Greer, CItr 17725, University of Idaho. Germplasm resources information network . Available at www.ars-grin.gov (accessed May 2014).Google Scholar
Radley, M. (1967) Site of production of gibberellin-like substances in germinating barley embryos. Planta 75, 164–171.CrossRefGoogle ScholarPubMed
Rikiishi, K. and Maekawa, M. (2010) Characterization of a novel wheat (Triticum aestivum L.) mutant with reduced seed dormancy. Journal of Cereal Science 51, 292–298.CrossRefGoogle Scholar
Ringlund, K. (1992) The importance of pre-harvest sprouting research. pp. 3–7 in Walker-Simmons, M.K.; Ried, J.L. (Eds) Pre-harvest sprouting in cereals. St. Paul, Minnesota, USA, American Association of Cereal Chemists.Google Scholar
Rodríguez, V.M., Margineda, M., González-Martín, J.F., Insausti, P. and Benech-Arnold, R.L. (2001) Predicting preharvest sprouting susceptibility in barley. Agronomy Journal 93, 1071–1079.CrossRefGoogle Scholar
Rodriguez, M.V., Barrero, J.M., Corbineau, F., Gubler, F. and Benech-Arnold, R.-L. (2015) Dormancy in cereals: not too much, not so little. Seed Science Research, in press .CrossRefGoogle Scholar
Schramm, E.C., Abellera, J.C., Strader, L.C., Campbell, K.G. and Steber, C.M. (2010) Isolation of ABA-responsive mutants in allohexaploid bread wheat (Triticum aestivum L.): drawing connections to grain dormancy, preharvest sprouting, and drought tolerance. Plant Science 179, 620–629.CrossRefGoogle Scholar
Schramm, E.C., Nelson, S.K. and Steber, C.M. (2012) Wheat ABA-insensitive mutants result in reduced grain dormancy. Euphytica 188, 35–49.CrossRefGoogle ScholarPubMed
Schramm, E.C., Nelson, S.K., Kidwell, K.K. and Steber, C.M. (2013) Increased ABA sensitivity results in higher seed dormancy in soft white spring wheat cultivar ‘Zak’. Theoretical and Applied Genetics 126, 791–803.CrossRefGoogle ScholarPubMed
Sears, R.G., Moffatt, J.M., Martin, T.J., Cox, T.S., Bequette, R.K., Curran, S.P., Chung, O.K., Heer, W.F., Long, J.H. and Witt, M.D. (1997) Registration of ‘Jagger’ wheat. Crop Science 37, 1010.CrossRefGoogle Scholar
Spielmeyer, W., Ellis, M., Robertson, M., Ali, S., Lenton, J.R. and Chandler, P.M. (2004) Isolation of gibberellin metabolic pathway genes from barley and comparative mapping in barley, wheat and rice. Theoretical and Applied Genetics 109, 847–855.CrossRefGoogle ScholarPubMed
Steber, C.M., Cooney, S.E. and McCourt, P. (1998) Isolation of the GA-response mutant sly1 as a suppressor of ABI1-1 in Arabidopsis thaliana . Genetics 149, 509–521.CrossRefGoogle ScholarPubMed
Syngenta . (2011) SY Ovation: New soft white winter wheat, AgriPro Syngenta. Available from http://washingtoncrop.com/wp-content/uploads/2012/12/SY-Ovation.pdf. https://agriprowheat.com/sites/default/files/file-attachments/sy_ovation_web.pdf. https://agriprowheat.com/wheat-variety/pacific-northwest/sy-ovation-0 (accessed 20 November 2014).Google Scholar
Wahl, T.I. and O'Rourke, D.A. (1993) The economics of sprout damage in wheat. pp. 10–17 in Walker-Simmons, M.K.; Reid, J.L. (Eds) Pre-harvest sprouting in cereals 1992. St. Paul, Minnesota, USA, American Association of Cereal Chemists.Google Scholar
Walker-Simmons, M. (1987) ABA levels and sensitivity in developing wheat embryos of sprouting resistant and susceptible cultivars. Plant Physiology 84, 61–66.CrossRefGoogle ScholarPubMed
Walker-Simmons, M. (1988) Enhancement of ABA responsiveness in wheat embryos by high temperature. Plant, Cell and Environment 11, 769–775.CrossRefGoogle Scholar
Warner, R.L., Kudma, D.A., Spaeth, S.C. and Jones, S.S. (2000) Dormancy in white-grain mutants of Chinese spring wheat (Triticum aestivum L.). Seed Science Research 10, 51–60.CrossRefGoogle Scholar
WestBred . (2014) WB-Arrowhead hard red winter, WestBred Northwest Region. Available from http://www.westbred.com/Products/Northwest/pages/showpdf.aspx?product =WB-ARROWHEAD (accessed 20 November 2014).Google Scholar
Wilkinson, M.D., McKibbin, R.S., Bailey, P.C., Flintham, J.E., Gale, M.D., Lenton, J.R. and Holdsworth, M.J. (2002) Use of comparative molecular genetics to study pre-harvest sprouting in wheat. Euphytica 126, 27–33.CrossRefGoogle Scholar
Yamauchi, Y., Ogawa, M., Kuwahara, A., Hanada, A., Kamiya, Y. and Yamaguchi, S. (2004) Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell 16, 367–378.CrossRefGoogle ScholarPubMed
Yoshimoto, K., Jikumaru, Y., Kamiya, Y., Kusano, M., Consonni, C., Panstruga, R., Ohsumi, Y. and Shirasu, K. (2009) Autophagy negatively regulates cell death by controlling NPR1-dependent salicylic acid signalling during senescence and the innate immune response in Arabidopsis. Plant Cell 21, 2914–2927.CrossRefGoogle ScholarPubMed
Zemetra, R.S., Lauver, M.L., O'Brien, K., Koehler, T., Souza, E.J., Guy, S.O., Robertson, L. and Brown, B. (2003) Registration of ‘Brundage 96’ wheat. Crop Science 43, 1884.CrossRefGoogle Scholar
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