Cereal Research Communications

, Volume 45, Issue 3, pp 466–477 | Cite as

SSR Marker Aided Introgression for opaque2 Allele for Development of Quality Protein Maize Inbreds

  • R. SinghEmail author
  • L. Ram
  • R. K. Singh
  • D. Singh Jakhar


Maize protein quality is deficit in essential amino acids, lysine and tryptophan. These constraints of o2 (opaque2) are corrected in genetically improved, hard endosperm QPM (Quality Protein Maize). An integrated strategy of phenotypic selection for endosperm modifiers and molecular marker-assisted foreground and background selection has been used in present study. The QPM donors were, CML 161, DMRQPM 58, CML 176 and CML 141 whereas, normal maize inbreds were CM 212, V338, V361, V336, V341, V351, CM 141 and V335. The inbreds were subjected to parental polymorphism survey between non-QPM and QPM using CIMMYT based three SSR markers, viz. phi057, umc1066 and phi112. Two markers, viz. phi057 and umc1066 exhibited co-dominant reactions, while phi112 was dominant in nature. Finally, two combinations V335 × CML 141 and V351 × CML 141 were considered for conversion program. Foreground selection was exercised using o2 specific marker umc1066 in BC1 and BC2 generations, while background as well as foreground selection was exercised in BC2F3 generation to recover the genome of recurrent parent up to extent of 80 to 100% with the help of SSR markers distributed across the whole maize genome. The tryptophan concentration in endosperm protein was significantly enhanced and the converted maize lines had almost twice the amount of lysine and tryptophan than normal maize inbreds.


background and foreground selection lysine opaque2 QPM tryptophan 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Frisch, M., Bohn, M., Melchinger, A.E. 1999. Comparison of selection strategies for marker assisted backcrossing of a gene. Crop Sci. 39:1295–1301.CrossRefGoogle Scholar
  2. Gupta, H.S., Babu, R., Agarwal, P.K., Mahajan, V., Hossian, F., Nepolean, T. 2014. Accelerated development of Quality Protein Maize hybrids through marker assisted introgression of opaque-2 allele. Plant Breed. 132:77–82.CrossRefGoogle Scholar
  3. Hernandez, H.H., Bates, L.S. 1969. A modified method for rapid tryptophan analysis in maize. CIMMYT Research Bulletin No. 13. CIMMYT. Mexico, D.F., Mexico.Google Scholar
  4. Hospital, F., Chevalet, C., Mulsant, P. 1992. Using markers in gene introgression programs. Genetics 132:1199–1210.PubMedPubMedCentralGoogle Scholar
  5. Krivanek, A., Groote, H., Gunaratna, N., Diallo, A., Freisen, D. 2007. Breeding and disseminating quality protein maize for Africa. Afr. J. Biotechnol. 6:312–324.Google Scholar
  6. Lopes, M.A., Larkins, B.A. 1995. Genetic analysis of opaque2 modifier gene activity in maize endosperm. Theor. Appl. Genet. 119:274–281.CrossRefGoogle Scholar
  7. Mertz, E.T., Bates, L.S., Nelson, O.E. 1964. Mutant gene that changes protein composition and increases lysine content of maize endosperm. Science 145:279–280.CrossRefGoogle Scholar
  8. Moro, G.L., Habben, J.E., Hamaker, B.R., Larkins, B.A. 1996. Characterization of the variability in lysine content for normal and opaque2 maize endosperm. Crop Sci. 36:1651–1659.CrossRefGoogle Scholar
  9. Pixley, K.V., Bjarnasan, M.S. 1993. Combining ability for yield and protein quality among modified endosperm opaque 2 tropical maize inbreds. Crop Sci. 33:1229–1234.CrossRefGoogle Scholar
  10. Prasanna, B.M., Vasal, S.K., Kasahun. B., Singh, N.N. 2001. Quality protein maize. Curr. Sci. 81:1308–1319.Google Scholar
  11. Ram, L. 2014. Improvement of non QPM lines through introgression of QPM genes. PhD Thesis, BHU. Varanasi, India.Google Scholar
  12. Ram, L., Singh, R., Singh, S.K. 2015. Study of combining ability using QPM donors as testers for yield and its component traits in maize (Zea mays L.). Sabrao J. Breeding and Genetics. 47:99–112.Google Scholar
  13. Ribaut, J.M., Hoisington, D.A. 1998. Marker assisted selection: new tools and strategies. Trends Plant Sci. 3:236–239.CrossRefGoogle Scholar
  14. Senior, M.L., Murphy, J.P., Goodman, M.M., Stuber, C.W. 1998. Utility of SSRs for determining genetic similarities and relationships in maize using an agarose gel system. Crop Sci. 38:1088–1098.CrossRefGoogle Scholar
  15. Singh, R., Srivastava, R.P. 2015. New quantitative trait loci (QTLs) for turcicum leaf blight in maize. J. of Biotech. Crop Sci. 5:35–43.Google Scholar
  16. Sofi, P.A., Wani, S.A., Rather, A.G., Wani, S.H. 2009. Quality protein maize (QPM): genetic manipulation for the nutritional fortification of maize. J. Plant Breed. Crop Sci. 1:244–253.Google Scholar
  17. Tanksley, S.D. 1993. Mapping polygenes. Annu. Rev. Genet. 27:205–233.CrossRefGoogle Scholar
  18. Vasal, S.K. 2001. High quality protein corn. In: Hallauer, A.R. (ed.), Specialty Corns. Second Edition. CRC Press. Boca Raton, FL, USA. pp. 85–129.Google Scholar
  19. Villegas, E., Mertz, T.E. 1975. Simple chemical and biological methods used at Purdue University to evaluate cereals for protein quality. St. Bull 70:17–21.Google Scholar
  20. Villegas, E., Ortega, E., Bauer, R. 1984. Chemical methods used at CIMMYT for determining protein quality in corn. CIMMYT. Mexico.Google Scholar
  21. Visscher, P.M., Haley, C.S., Thompson, R. 1996. Marker assisted introgression in backcross breeding programs. Genetics 144:1923–1932.PubMedPubMedCentralGoogle Scholar
  22. Young, N.D., Tanksley, S.D. 1989. RFLP analysis of the size of chromosomal segments retained around “Tm-2 locus” of tomato during backcross breeding. Theor. Appl. Genet. 77:353–359.CrossRefGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2017

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • R. Singh
    • 1
    Email author
  • L. Ram
    • 1
  • R. K. Singh
    • 2
  • D. Singh Jakhar
    • 1
  1. 1.Department of Genetics and Plant Breeding, Institute of Agricultural SciencesBanaras Hindu UniversityVaranasi-Uttar PradeshIndia
  2. 2.Visiting Scientist, Plant Genome Mapping LaboratoryUniversity of GeorgiaAthens, GAUSA

Personalised recommendations