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Molecular Marker Analysis of Protein Content Using PCR-Based Markers in Wheat

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Abstract

Grain protein concentration (GPC) of hexaploid wheat is one of the important factors that determines the end-product quality, as well as playing a pivotal role in human nutrition. In an attempt to identify PCR-based DNA markers linked to GPC, 106 recombinant inbred lines (RILs) were developed from a cross between two wheat cultivars PH132 and WL711, which differ significantly in GPC, by the single seed descent method. The RILs were phenotyped for GPC at two diverse agroclimatic locations, namely Pune and Ludhiana, to study the influence of genotype and environment interactions on this trait. The parents were screened with 85 inter simple sequence repeat (ISSR) primers and 350 random primers. The selective genotyping and whole population analysis revealed nine DNA markers associated with the trait. Three markers (UBC8441100, UBC8801100, and OPA4800) were observed to be associated with the trait in both locations, whereas two markers (OPH41400 and UBC873750) were found to be specific to Pune, and four markers (OPM5870, OPO10870, OPV141200, and UBC8251000) were specific to Ludhiana. Together five markers at the Pune location representing five QTLs and seven markers at Ludhiana representing four QTLs accounted for 13.4 and 13.5% of total phenotypic variation, respectively. This study clearly demonstrates that GPC is highly influenced by the environment, and the applicability of ISSR and RAPD markers in finding regions on chromosomes associated with quantitative characters in wheat such as GPC.

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REFERENCES

  • AACC (1983). Approved Methods, 8th edn. American Association of Cereal Chemistry, St. Paul, MN.

    Google Scholar 

  • Akagi, H., Yokozoki, Y., Inagaki, A., Nakamura, A., and Fugimura, T. (1996). A codominant DNA marker closely linked to the rice nuclear restorer gene, Rf-1, identified with inter simple sequence repeat. Genome 39:1205-1209.

    Google Scholar 

  • Ammiraju, J. S. S., Dholakia, B. B., Santra, D. K., Singh, H., Lagu, M. D., Tamhankar, S. A., Dhaliwal, H. S., Rao, V. S., Gupta, V. S., and Ranjekar, P. K. (2001). Identification of inter simple sequence repeat (ISSR) markers associated with seed size in wheat. Theor: Appl. Genet. 102:726-732.

    Google Scholar 

  • Austin, D. F., and Lee, M. (1998). Determination of quantitative trait loci for grain yield and yield components in maize across generations in stress and non stress environments. Crop Sci. 38:1296-1308.

    Google Scholar 

  • Barneix, A. J., Fatta, N., Kade, M., Pfluger, L., and Suarez, E. Y. (1998). Effect of wheat chromosome 7BS on grain protein concentration. Cereal Res. Commun. 26:101-106.

    Google Scholar 

  • Bhatt, G. M., and Derera, N. F. (1975). Genotype × environment interactions for heritabilities and correlations among quality traits in wheat. Euphytica 24:597-604.

    Google Scholar 

  • Blanco, A., Giovanni, de C., Laddomada, B., Sciancalepore, A., Simeone, R., Devos, K. M., and Gale, M. D. (1996). Quantitative trait loci influencing grain protein content in tetraploid wheat. Plant Breed. 115:310-316.

    Google Scholar 

  • Campbell, K. G., Bergman, C. J., Gaulberto, D. G., Anderson, J. A., Giroux, M. J., Hareland, G., Gulcher, R. G., Sorrells, M. E., and Finney, P. L. (1999). Quantitative trait associated with kernel traits in a soft × hard wheat cross. Crop Sci. 39:1184-1195.

    Google Scholar 

  • Cox, M. C., Qualset, C. O., and Rains, D. W. (1985). Genetic variation for nitrogen assimilation and translocation in wheat. II. Nitrogen assimilation in relation to grain yield and protein. Crop Sci. 25:430-435.

    Google Scholar 

  • Darvasi, M., and Soller, M. (1992). Selective genotyping for determination of linkage between a marker locus and a quantitative trait locus. Theor: Appl. Genet. 85:357-359.

    Google Scholar 

  • Day, G. E., Paulsen, G. N., and Sears, R. G. (1985). Nitrogen relations in winter wheat cultivars differing in grain protein percentage and structure. J. Plant Nutr. 8:555-566.

    Google Scholar 

  • Dhaliwal, L. S., Harjit, S., Nanda, G. S., and Dhaliwal, H. S. (1994). Inheritance of grain protein content in two high protein lines of wheat. Rachis 13:34-37.

    Google Scholar 

  • Diehl, A. L., Johnson, V. A., and Mattern, P. J. (1978). Inheritance of protein and lysine in three wheat crosses. Crop Sci. 17:391-395.

    Google Scholar 

  • Dong, H., Cox, T. S., Sears, R. G., and Lookhart, G. L. (1991). High molecular weight glutenin genes: Effect on quality in wheat. Crop Sci. 31:974-979.

    Google Scholar 

  • Goldman, I. L., Rocheford, T. R., and Dudley, J. W. (1993). Quantitative trait loci influencing protein and starch concentration in the Illinois term selection maize strains. Theor: Appl. Genet. 87:217-224.

    Google Scholar 

  • Gonzalez-Hernandez, J. L., Kianian, S., and Elias, E. M. (1998). Grain protein content and 1000-kernel weight QTL mapping on chromosome 5B of Triticum dicoccoides. In Slinkard AE (ed.), Proceedings of the Ninth International Wheat Genetic Symposium, Saskatoon, Saskatchewan, Canada, Aug. 2–7, 1998, pp. 47-49.

  • Halloran, G. M. (1975). Genetic analysis of grain protein percentage in wheat. Theor: Appl. Genet. 46:79-86.

    Google Scholar 

  • Johnson, V. A., and Mattern, P. J. (1987). Wheat, Rye and triticale. In Heine EG (ed.), Nutritional Quality of Cereal Grains: Genetic and Agronomic Improvement, ASA-CSSA-SSSA, Madison, WI. Agronomy Monograph, Vol. 28, pp. 133-182.

    Google Scholar 

  • Joppa, L. R., and Cantrell, R. G. (1990). Chromosomal location of genes for grain protein content of wild tetraploid wheat. Crop Sci. 30:1059-1064.

    Google Scholar 

  • Joppa, L. R., Changheng, D., Hart, G. E., and Hareland, G. A. (1997). Mapping gene(s) for grain protein in tetraploid wheat (Triticum turgidum L.) using a population of recombinant inbred chromosome lines. Crop. Sci. 37:1586-1589.

    Google Scholar 

  • Kojima, T., Nagaoka, T., Noda, K., and Ogihara, Y. (1998). Genetic linkage map of ISSR and RAPD markers in Einkorn wheat in relation to that of RELP markers. Theor: Appl. Genet. 96:37-45.

    Google Scholar 

  • Konzak, C. F. (1977). Genetic control of the content aminoacid composition and processing properties of proteins in wheat. Adv. Genet. 19:407-582.

    Google Scholar 

  • Kuspira, J., and Unrau, J. (1957). Genetic analysis of certain characters in common wheat using whole chromosome substitution lines. Can. J. Plant Sci. 37:300-326.

    Google Scholar 

  • Lande, R., and Thompson, R. (1990). Efficiency of marker assisted selection in the improvement of quantitative traits. Genetics 124:743-756.

    Google Scholar 

  • Lander, E. S., and Botstein, D. (1989). Mapping mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121:185-199.

    Google Scholar 

  • Lander, E. S., Green, P., Abrahamson, J., Barlow, A., Day, M. J., Lincoln, S. E., and Newburg, L. (1987). MAPMAKER: An interacive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174-181.

    Google Scholar 

  • Leavy, A. A., Zaccai, M., Millet, E., and Feldman, M. (1988). Utilization of wild emmer for the improvement of grain protein percentage of cultivated wheat. In Miller, T. E., and Koebner, R. M. (eds.), Proceedings of the Seventh Wheat Genetics Symposium, Cambridge July 13–19, 1988, Institute of Plant Science Research Cambridge Lab, Cambridge, England, pp. 969-974.

    Google Scholar 

  • Lee, S. H., Bailey, M. A., Mian, M. A. R., Carter, T. E. Jr., Shipe, E. R., Ashley, D. A., Parrott, W. A., Hussey, R. S., and Boerma, H. R. (1996). RFLP loci associated with soybean seed protein and oil content across populations and locations. Theor: Appl. Genet. 93:649-657.

    Google Scholar 

  • Lee, S. J., and Penner, G. A. (1997). The conversion of RFLP markers to allele specific amplicons linked to QTLs governing malting quality in barley. Mol. Breed. 3:457-462.

    Google Scholar 

  • Lu, C., Shen, L., Tan, Z., Xu, Y., He, P., Chen, Y., and Zhu, L. (1997). Comparative mapping of QTLs for agronomic traits of rice across environments by using a doubled-haploid population. Theor: Appl. Genet. 94:145-150.

    Google Scholar 

  • Martin, G. B., Williams, J. G. K., and Tanksley, S. D. (1991). Rapid identification of markers linked to a pseudomonas resistance gene in tomato by using random primers and near isogenic lines. Proc. Natl. Acad. Sci. (USA) 88:2336-2340.

    Google Scholar 

  • Mesfin, A., Frohberg, R. C., Khan, K., and Olson, T. C. (2000). Increased grain protein content and its association with agronomic and end-use quality in two hard red spring wheat populations derived from Triticum turgidum L. var. dicoccoides. Euphytica 116:237-242.

    Google Scholar 

  • Moretzson, M. C., Nunes, C. D. M., Ferreira, M. E., and Grattapaglia, D. (2000). RAPD linkage mapping of the shell thickness locus in oil palm (Elaeis guineensis Jacq.) Theor: Appl. Genet. 100:63-70.

    Google Scholar 

  • Morris, R., Mattern, P. J., Schmidt, J. W., and Johnson, V. A. (1978). Studies on protein, lysine and leaf rust resistance in the wheat cultivar “Atlas 66” using chromosome substitutions. In Ramanujan, S. (ed.), Proceedings of the Fifth International Wheat Genetic Symposium, Indian Agricultural Research Institute, New Delhi, pp. 447-454.

    Google Scholar 

  • Mulitze, D. K. (1998). Analysis of wheat breeding data using AGROBASE™ 98 for windows. In Slinkard, A. E. (ed.), Proceedings of the Ninth International Wheat Genetic Symposium, Saskatoon, Saskatchewan, Canada, Aug. 2–7, 1998, pp. 295-297.

  • Muller, M., Muth, J. R., Gallansci, P., Knudsen, S., Maddalow, M., Motto, M., Schmitz, D., Sorenson, M. B., Salamani, F., Wettstein, D., and Thompson, R. D. (1995). Regulation of storage protein synthesis in cereal seeds: Developmental and nutritional aspects. J. Plant Physiol. 145:606-613.

    Google Scholar 

  • Nagaoka, T., and Ogihara, Y. (1997). Applicability of inter-simple sequence repeat polymorphism in wheat for use as DNA markers in comparison to RFLP and RAPD markers. Theor: Appl. Genet. 94:597-602.

    Google Scholar 

  • Naik, S., Gill, K. S., Rao, V. S., Gupta, V. S., Tamhankar, S. A., Pujar, S., Gill, B. S., and Ranjekar, P. K. (1998). Identification of STS marker linked to the Aegilops speltoides-derived leaf rust resistance gene Lr 28 in wheat. Theor. Appl. Genet. 97:535-540.

    Google Scholar 

  • Nelson, J. C. (1997). QGENE: Software for marker based genomic analysis and breeding. Mol. Breed. 3:359-366.

    Google Scholar 

  • Paterson, A. H., Damon, S., Hewitt, J. D., Zamir, D., Robinowitch, H. D., Lincoln, S. E., Lander, E. S., and Tanksley, S. D. (1991). Mendelian factors underlying quantitative traits in tomato: Comparison across species generations and environments. Genetics 127:181-197.

    Google Scholar 

  • Penner, G. A., Clarke, J., Bezte, L. J., and Leisle, D. (1995). Identification of RAPD markers linked to a gene governing cadmium uptake in durum wheat. Genome 38:543-547.

    Google Scholar 

  • Prasad, M., Varshney, R. K., Kumar, A., Balyan, H. S., Sharma, P. C., Edvard, K. J., Singh, H., Dhaliwal, H. S., Roy, J. K., and Gupta, P. K. (1999). A microsetellite marker associated with a QTL for GPC on chromosome arm 2DL of bread wheat. Theor. Appl. Genet. 99:341-345.

    Google Scholar 

  • Pujar, S., Tamhankar, S. A., Rao, V. S., Gupta, V. S., Naik, S., and Ranjekar, P. K. (1999). Arbitrarily primed-PCR based diversity assessment reflects hierarchical groupings of Indian tetraploid wheat genotypes. Theor. Appl. Genet. 99:868-876.

    Google Scholar 

  • Rantanparkhe, M. B., Santra, D. K., Tullu, A., and Muehlbauer, F. J. (1998). Inherince of inter-simple-sequence-repeat polymorphism and linkage with a fusarium wilt resistance gene in chickpea. Theor. Appl. Genet. 96:348-353.

    Google Scholar 

  • Rogers, S. O., and Bendich, A. J. (1988). Extraction of DNA from plant tissue. In Gelvin S. B., Shilperoort, R. A. (eds.). Plant Molecular Biology Manual, Kluwer Academic, Dordrecht, pp. 1-10.

    Google Scholar 

  • Sears, E. R. (1996). Nullisomic-tetrasomic combinations in hexaploid wheat. In Riley, R., and Lewis, E. R. (ed.), Chromosome Manipulation and Plant Genetics, Oliver and Boyd, Edinburg, pp. 29-45.

    Google Scholar 

  • Sears, E. R., and Sears, L. M. S. (1978). The telocentric chromosomes of common wheat. In Ramanujan, S. (ed.), Fifth International Wheat Genetic Symposium, New Delhi, India Feb. 23–28, 1978, Indian Society of Genetics and Plant Breeding, Indian Agricultural Research Institute, New Delhi. pp. 389-4070.

    Google Scholar 

  • Sourdille, P., Perretarrt, M. R., Charmet, G., Leroy, P., Gantier, M. F., Joudier, P., Nelson, J. C., Sorrells, M. E., and Bernard, M. (1996). Linkage between RFLP markers and genes affecting kernel hardness in wheat. Theor. Appl. Genet. 93:580-586.

    Google Scholar 

  • Stein, I. S., Sears, R. G., Gill, B. S., Hoseney, R. C., and Cox T. S. (1992). Heterogeneity of the “Wichita” wheat monosomic set for grain quality and agronomic traits. Crop Sci. 32:581-584.

    Google Scholar 

  • Stuber, C. W., Edwards, M. D., and Wendel, J. F. (1987). Molecular marker facilitated investigation of quantitative trait loci in maize. II Factors influencing yield and its component traits. Crop Sci. 27:639-648.

    Google Scholar 

  • Tan, Y. F., Li, J. X., Yu, S. B., Xing, Y. Z., Xu, C. G., and Zhang, Q. (1999). The three important traits for cooking and eating quality of rice grains are controlled by a single locus in an elite rice hybrid, Shanyou 63. Theor. Appl. Genet. 99:642-648.

    Google Scholar 

  • Udall, J. A., Souza, E., Anderson, J., Sorrells, M. E., and Zemetra, R. S. (1999). Quantitative traits loci for flour viscosity in winter wheat. Crop Sci. 39:238-242.

    Google Scholar 

  • Zietkiewicz, E., Rafalski, A., and Labuda, D. (1994). Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176-183.

    Google Scholar 

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Authors and Affiliations

  1. Plant Molecular Biology Unit, Division of Biochemical Sciences, National Chemical Laboratory, Pune, Maharashtra, India

    B. B. Dholakia, J. S. S. Ammiraju, D. K. Santra, M. V. Katti, M. D. Lagu, V. S. Gupta & P. K. Ranjekar

  2. Biotechnology Center, Punjab Agricultural University, Ludhiana, Punjab, India

    H. Singh & H. S. Dhaliwal

  3. Genetics and Plant Breeding Unit, Division of Plant Sciences, Agharkar Research Institute, Maharashtra, India

    S. A. Tamhankar & V. S. Rao

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  1. B. B. Dholakia
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  2. J. S. S. Ammiraju
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  3. D. K. Santra
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  4. H. Singh
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  5. M. V. Katti
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  6. M. D. Lagu
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  7. S. A. Tamhankar
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  8. V. S. Rao
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  9. V. S. Gupta
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  10. H. S. Dhaliwal
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  11. P. K. Ranjekar
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Dholakia, B.B., Ammiraju, J.S.S., Santra, D.K. et al. Molecular Marker Analysis of Protein Content Using PCR-Based Markers in Wheat. Biochem Genet 39, 325–338 (2001). https://doi.org/10.1023/A:1012256813965

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