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Creation and functional analysis of new Puroindoline alleles in Triticum aestivum

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Abstract

The Hardness (Ha) locus controls grain texture and affects many end-use properties of wheat (Triticum aestivum L.). The Ha locus is functionally comprised of the Puroindoline a and b genes, Pina and Pinb, respectively. The lack of Pin allelic diversity is a major factor limiting Ha functional analyses and wheat quality improvement. In order to create new Ha alleles, a 630 member M2 population was produced in the soft white spring cultivar Alpowa using ethylmethane sulfonate mutagenesis. The M2 population was screened to identify new alleles of Pina and Pinb. Eighteen new Pin alleles, including eight missense alleles, were identified. F2 populations for four of the new Pin alleles were developed after crossing each back to non-mutant Alpowa. Grain hardness was then measured on F2:3 seeds and the impact of each allele on grain hardness was quantified. The tested mutations were responsible for between 28 and 94% of the grain hardness variation and seed weight and vigor of all mutation lines was restored among the F2 populations. Selection of new Pin alleles following direct phenotyping or direct sequencing is a successful approach to identify new Ha alleles useful in improving wheat product quality and understanding Ha locus function.

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References

  • Anderson P (1995) Mutagenesis methods. Cell Biol 48:31–58

    Article  CAS  Google Scholar 

  • Bagge M, Xia X, Lübberstedt T (2007) Functional markers in wheat. Curr Opin Plant Biol 102:211–216

    Article  Google Scholar 

  • Beecher B, Bettge A, Smidansky E, Giroux MJ (2002) Expression of wild-type pinB sequence in transgenic wheat complements a hard phenotype. Theor Appl Genet 105:870–877

    Article  PubMed  CAS  Google Scholar 

  • Blochet JE, Chevalier C, Forest E, Pebay-Peyroula E, Gautier MF, Joudrier P, Pezolet M, Marion D (1993) Complete amino acid sequence of Puroindoline, a new basic and cystine-rich protein with a unique tryptophan-rich domain, isolated from wheat endosperm by Triton X-114 phase partitioning. FEBS Lett 329:336–340

    Article  PubMed  CAS  Google Scholar 

  • Caldwell DG, McCallum N, Shaw P, Muehlbauer GJ, Marshall DF, Waugh RA (2004) A structured mutant population for forward and reverse genetics in barley (Hordeum vulgare L). Plant J 40:143–150

    Article  PubMed  CAS  Google Scholar 

  • Campbell KG, Bergman CJ, Gualberto DG, Anderson JA, Giroux MJ, Hareland G, Fulcher G, Sorrels ME, Finney PL (1999) Quantitative trait loci associated with kernel traits in a soft hard wheat cross. Crop Sci 39:11841195

    Google Scholar 

  • Campbell JB, Martin JM, Crutcher F, Meyer FD, Clark DR, Giroux MJ (2007) Effects on soft wheat (Triticum aestivum L.) quality of increased puroindoline dosage. Cereal Chem 84(1):80–87

    Google Scholar 

  • Chang C, Zhang H, Xu J, Li W, Liu G, You M, Li B (2006) Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE. Euphytica 152:225–234

    Article  CAS  Google Scholar 

  • Chen F, He ZH, Xia XC, Lillemo M, Morris C (2005) A new puroindoline b mutation present in Chinese winter wheat cultivar Jingdong 11. J Cereal Sci 42:267–269

    Article  CAS  Google Scholar 

  • Chen F, He ZH, Xia XC, Xia LQ, Zhang XY, Lillemo M, Morris CF (2006) Molecular and biochemical characterization of Puroindoline a and b alleles in Chinese landraces and historical cultivars. Theor Appl Genet 112:400–409

    Article  PubMed  CAS  Google Scholar 

  • Colbert T, Till BJ, Tompa R, Reynolds S, Steine MN, Yeung AT, McCallum CM, Comai l, Henikoff S (2001) High-throughput screening for induced point mutations. Plant Physiol 126:480–484

    Article  PubMed  CAS  Google Scholar 

  • Cox TS (1998) Deepening the wheat gene pool. J Crop Prod 1:1–25

    Article  Google Scholar 

  • Feiz L, Martin JM, Giroux MJ (2008) The relationship between wheat (Triticum aestivum L.) grain hardness and wet-milling quality. Cereal Chem 85(1):44–50

    Google Scholar 

  • Gautier MF, Aleman ME, Guirao A, Marion D, Joudrier P (1994) Triticum aestivum Puroindolines, two basic cystine-rich seed proteins: cDNA sequence analysis and developmental gene expression. Plant Mol Biol 25:43–57

    Article  PubMed  CAS  Google Scholar 

  • Gedye KR, Morris CF, Bettge AD (2004) Determination and evaluation of the sequence and textural effects of the Puroindoline a and Puroindoline b genes in a population of synthetic hexaploid wheat. Theor Appl Genet 109:1597–1603

    Article  PubMed  CAS  Google Scholar 

  • Gill BS, Raupp WJ (1987) Direct genetic transfers from Aegilops squarrosa L to hexaploid wheat. Crop Sci 27:445–450

    Google Scholar 

  • Giroux MJ, Morris CF (1997) A glycine to serine change in Puroindoline b is associated with wheat grain hardness and low levels of starch-surface friabilin. Theor Appl Genet 95:857–864

    Article  CAS  Google Scholar 

  • Giroux MJ, Morris CF (1998) Wheat grain hardness results from highly conserved mutations in the friabilin components Puroindoline a and b. Proc Natl Acad Sci USA 95:6262–6266

    Article  PubMed  CAS  Google Scholar 

  • Giroux MJ, Talbert L, Habernicht DK, Lanning S, Hemphill A, Martin JM (2000) Association of puroindoline sequence type and grain hardness in hard red spring wheat. Crop Sci 40:370–374

    CAS  Google Scholar 

  • Greenblatt GA, Bettge AD, Morris CF (1995) The relationship among endosperm texture, friabilin occurrence, and bound polar lipids on wheat starch. Cereal Chem 72:172–176

    CAS  Google Scholar 

  • Greene EA, Codomo CA, Taylor NE, Henikoff JG, Till BJ, Reynolds SH, Enns LC, Burtner C, Johnson JE, Odden AR, Comai L, Henikoff S (2003) Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. Genetics 164:731–740

    PubMed  CAS  Google Scholar 

  • Greenwell P, Schofield JD (1986) A starch granule protein associated with endosperm softness in wheat. Cereal Chem 63:379–380

    CAS  Google Scholar 

  • Hogg AC, Sripo T, Beecher B, Martin JM, Giroux MJ (2004) Wheat Puroindolines interact to form friabilin and control wheat grain hardness. Theor Appl Genet 108:1089–1097

    Article  PubMed  CAS  Google Scholar 

  • Hogg AC, Beecher B, Martin JM, Meyer F, Talbert L, Lanning S, Giroux MJ (2005) Hard wheat milling and bread baking traits affected by the seed-specific overexpression of Puroindolines. Crop Sci 45(3):871–878

    Article  CAS  Google Scholar 

  • Jander G, Baerson SR, Hudak JA, Gonzalez KA, Gruys KJ, Last RL (2003) Ethylmethanesulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. Plant Physiol 131:139–146

    Article  PubMed  CAS  Google Scholar 

  • Kooijman M, Orsel R, Hessing M, Hamer RJ, Bekkers ACAPA (1997) Spectroscopic characterization of the lipid-binding properties of wheat puroindolines. J Cereal Sci 26:145–159

    Article  CAS  Google Scholar 

  • Krishnamurthy K, Giroux MJ (2001) Expression of wheat puroindoline genes in transgenic rice confers grain softness. Nature Biotech 19:162–166

    Article  CAS  Google Scholar 

  • Krishnamurthy K, Balconi C, Sherwood JE, Giroux MJ (2001) Increased tolerance to fungal diseases of rice plants transformed with puroindoline genes. Mol Plant Microbe Interact 14:1255–1260

    Article  PubMed  CAS  Google Scholar 

  • Law CN, Young CF, Brown JWS, Snape JW, Worland JW (1978) The study of grain protein control in wheat using whole chromosome substitution lines In: Seed protein improvement by nuclear techniques. International Atomic Energy Agency, Vienna, Austria, pp 483–502

    Google Scholar 

  • Lillemo M, Morris CF (2000) A leucine to proline mutation in Puroindoline b is frequently present in hard wheats from Northern Europe. Theor Appl Genet 100:1100–1107

    Article  CAS  Google Scholar 

  • Marion D, Gautier MF, Joudrier P, Ptak M, Pezolet M, Forest E, Clark DC Broekaert W (1994) Structure and function of wheat lipid binding proteins. In: Martino al Cimino S (ed) Wheat kernel proteins—molecular and functional aspects, pp 175–180, Universita degli studi della tuscia Viterbo, Italy

  • Martin CR, Rousser R, Brabec DL (1993) Development of a single-kernel wheat characterization system. Trans ASAE 36:1399–1404

    Google Scholar 

  • Martin JM, Frohberg RC, Morris CF, Talbert LE, Giroux MJ (2001) Milling and bread baking traits associated with puroindoline sequence type in hard red spring wheat. Crop Sci 41:228–234

    CAS  Google Scholar 

  • Martin JM, Meyer FD, Smidansky ED, Wanjugi H, Blechl AE, Giroux MJ (2006) Complementation of the pina (null) allele with the wild type Pina sequence restores a soft phenotype in transgenic wheat. Theor Appl Genet 113:1563–1570

    Article  PubMed  CAS  Google Scholar 

  • Martin JM, Meyer FD, Morris CF, Giroux MJ (2007) Pilot scale milling characteristics of transgenic isolines of a hard wheat over-expressing puroindolines. Crop Sci 47(2):497–504

    Google Scholar 

  • Martin JM, Sherman JD, Lanning SP, Talbert LE, Giroux MJ (2008) Effect of variation in amylose content and puroindoline composition on bread quality in a hard spring wheat population. Cereal Chem 85(2):266–269

    Article  CAS  Google Scholar 

  • Massa AN, Morris CF, Gill BS (2004) Sequence diversity of puroindoline-a, puroindoline-b, and the grain softness protein genes in Aegilops tauschii cross. Crop Sci 44:1808–1816

    CAS  Google Scholar 

  • Mattern PJ, Morris R, Schmidt JW, Johnson VA (1973) Location of genes for kernel properties in the wheat variety ‘Cheyenne’ using chromosome substitution lines In: Proceedings of the 4th international wheat genetics symposium. Columbia, MO, pp 703–707

    Google Scholar 

  • McIntosh RA, Devos KM, Dubcovsky J, Rogers WJ, Morris CF, Appels R, Somers DJ, Anderson OA (2007) Catalogue of gene symbols for wheat: 2007 supplement. http://www.grain.jouy.inra.fr/ggpages/awn/53Textfile/WGC.html

  • Morris CF, King GE (2008) Registration of hard kernel puroindoline allele near-isogenic line hexaploid wheat genetic stocks. J Plant Reg 2:67–68

    Article  Google Scholar 

  • Morris CF, Rose SP (1996) Wheat. In: Henry RJ, Kettlewell PS (eds) Cereal grain quality. Chapman & Hall, London, pp 3–54

    Google Scholar 

  • Morris CF, Greenblatt GA, Bettge AD, Malkawi HI (1994) Isolation and characterization of multiple forms of friabilin. J Cereal Sci 21:167–174

    Article  Google Scholar 

  • Morris CF, Lillemo M, Simeone MC, Giroux MJ, Babb SL, Kidwell KK (2001) Prevalence of Puroindoline grain hardness genotypes among historically significant North American spring and winter wheats. Crop Sci 41:218–228

    CAS  Google Scholar 

  • Ng PC, Henikoff S (2003) SIFT: predicting amino acid changes that affect protein function. Nuc Acid Res 31:3812–3814

    Article  CAS  Google Scholar 

  • Pan Z, Song W, Meng F, Xu L, Liu B, Zhu J (2004) Characterization of genes encoding wheat grain hardness from Chinese cultivar GaoCheng 8901. Cereal Chem 81:287–289

    Article  CAS  Google Scholar 

  • Ram S, Boyko E, Giroux MJ, Gill BS (2002) Null mutation in Puroindoline A is prevalent in Indian wheats: Puroindoline genes are located in the distal part of short arm of 5D chromosome. J Plant Biochem Biotech 11:1–5

    Google Scholar 

  • Riede CR, Anderson JA (1996) Linkage of RFLP markers to an aluminum tolerance gene in wheat. Crop Sci 36:905–909

    Google Scholar 

  • SAS Institute, Inc. (2004) SAS/STAT 91 User’s guide SAS Institute Inc., Cary

  • Simeone MC, Gedye KR, Mason-Gamer R, Gill BS, Morris CF (2006) Conserved regulatory elements identified from a comparative puroindoline gene sequence survey of Triticum and Aegilops diploid taxa. J Cereal Sci 44:21–33

    Article  CAS  Google Scholar 

  • Slade AJ, Fuerstenberg SI, Loeffler D, Steine MN, Facciotti D (2005) A reverse genetic, nontransgenic approach to wheat crop improvement by TILLING. Nat Biotechnol 23:75–81

    Article  PubMed  CAS  Google Scholar 

  • Swan CG, Meyer FD, Hogg AC, Martin JM, Giroux MJ (2006) Puroindoline B limits binding of Puroindoline A to starch and grain softness. Crop Sci 46(4):1656–1665

    Article  CAS  Google Scholar 

  • Till BJ, Cooper J, Tai TH, Colowit P, Greene EA, Henikoff S, Comai L (2007) Discovery of chemically induced mutations in rice by TILLING. BMC Plant Biol 11:7–19

    Google Scholar 

  • Thorgeirsson TE, Russel CJ, King DS, Shin YK (1996) Direct determination of the membrane affinities of individual amino acids. Biochemistry 35:1803–1809

    Article  PubMed  CAS  Google Scholar 

  • Wanjugi HW, Hogg AC, Martin JM, Giroux MJ (2007a) The role of puroindoline A and B individually and in combination on grain hardness and starch association. Crop Sci 47:67–76

    Article  CAS  Google Scholar 

  • Wanjugi HW, Martin JM, Giroux MJ (2007b) Influence of Puroindolines A and B individually and in combination on wheat milling and bread traits. Cereal Chem 54(6):540–547

    Article  Google Scholar 

  • Warburton ML, Crossa J, Franco J, Kazi M, Trethowan R, Rajaram S, Pfeiffer W, Zhang P, Dreisigacker S, van Ginkel M (2006) Bringing wild relatives back into the family: recovering genetic diversity in CIMMYT improved wheat germplasm. Euphytica 149:289–301

    Article  CAS  Google Scholar 

  • Zhu JK, Liu J, Xiong L (1998) Genetic analysis of salt tolerance in Arabidopsis: evidence for a critical role of potassium nutrition. Plant Cell 10:1181–1192

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Plant Sciences and Plant Pathology, Montana State University, 119 Plant Bioscience Building, Bozeman, MT, 59717-3150, USA

    L. Feiz, J. M. Martin & M. J. Giroux

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  1. L. Feiz
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  2. J. M. Martin
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  3. M. J. Giroux
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Corresponding author

Correspondence to M. J. Giroux.

Additional information

Communicated by A. Graner.

This research was supported by USDA-ARS National Research Initiative Competitive Grants Program, grants 2004-35301-14538, 2007-35301-18135 and by the Montana Agricultural Experiment Station.

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Feiz, L., Martin, J.M. & Giroux, M.J. Creation and functional analysis of new Puroindoline alleles in Triticum aestivum . Theor Appl Genet 118, 247–257 (2009). https://doi.org/10.1007/s00122-008-0893-1

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  • DOI: https://doi.org/10.1007/s00122-008-0893-1

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