Molecular Interventions for Enhancing the Protein Quality of Maize

  • Yogesh Vikal
  • J. S. Chawla


Maize, one of the three most popular cereal crops of the world, globally contributes 15 % of the protein and 20 % of the calories derived from food crops in the world’s diet. However, cereals do not provide a nutritionally balanced source of protein. For nutritional security, it is necessary to adopt a genetic enhancement strategy in which essential amino acids are either incorporated or increased in grain protein to alleviate hunger, increase income, and improve livelihood. Quality protein maize (QPM) is having high nutritive value of endosperm protein with opaque2 (o2) mutation leading to 60–100 % increased content of lysine and tryptophan. The lysine value of o2 maize is 2.5–4.0 g/100 g of endosperm protein, which is more than twice that of the normal maize (1.3 g lysine/100 g protein). International Maize and Wheat Improvement Center (CIMMYT), Mexico, played a significant role in the development of QPM maize. The breeding of QPM involves three genetic systems: (i) the recessive mutant allele of the o2 gene, (ii) the endosperm hardness modifier genes, and (iii) the amino acid modifier genes influencing free amino acid content in the endosperm. Due to recessive nature of the o2 gene, complex action of modifier genes, and presence of amino acid enhancer genes, the use of DNA marker-assisted selection (MAS) accelerated the selection efficiency and expedited the development of new QPM cultivars. Using a combination of MAS and phenotypic selection techniques, a single cross, short duration Vivek QPM 9 hybrid was developed and released in 2008 by Vivekananda Parvatiya Krishi Anusandhan Sansthan, Almora, India. Alternatively, manipulating the plant lysine metabolic pathway provides possible enzyme targets for genetic engineering to increase free lysine content in corn grain. Furthermore, RNA interference (RNAi) has been used to specifically suppress α-zein production in transgenic corn, resulting in a doubling of the lysine content of corn grain. QPM is likely to gain wider acceptance if QTLs for kernel modification, and enhancers for amino acids are fine mapped to develop markers to follow MAS for vitreous kernels and high levels of lysine. However, the major constraints in adoption of QPM hybrids are contamination with normal maize pollen in field, resulting in erosion of the trait in farmer-saved seed system. It is essential to give training on good seed production practices to the local communities and development of linkage between the seed producers, farmers, and the industry for sustainable higher nutritional benefits of QPM in the long term.


Lysine Content Quality Protein Maize Nutritional Security Normal Maize Recurrent Parent Genome 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Agrawal PK, Gupta HS (2010) Enhancement of protein quality of maize using biotechnological options. Anim Nutr Feed Tech 10S:79–91Google Scholar
  2. Azevedo RA, Lancien M, Lea PJ (2006) The aspartic acid metabolic pathway, an exciting and essential pathway in plants. Amino Acids 30:143–162PubMedCrossRefGoogle Scholar
  3. Babu R, Nair SK, Kumar A, Venkatesh S, Sekhar JC, Singh NN, Srinivasan G, Gupta HS (2005) Two-generation marker-aided backcrossing for rapid conversion of normal maize lines to quality protein maize. Theor Appl Genet 111:888–897PubMedCrossRefGoogle Scholar
  4. Bass HW, Webster C, O’Brien GR, Roberts JKM, Bostou RS (1992) A maize ribosome inactivating protein is controlled by the transcriptional activator opaque 2. Plant cell 4:225–234PubMedGoogle Scholar
  5. Bhatia CR, Rabson R (1987) Relationship of grain yield and nutritional quality. In: Olson RA, Frey KJ (eds) Nutritional quality of cereal grains: genetic and agronomic improvement, vol 28, Agronomy monograph. ASA, CSSA and SSSA, Madison, pp 11–43Google Scholar
  6. Bressani R (1992) Nutritional value of high-lysine maize in humans. In: Mertz ET (ed) Quality protein maize. American Association of Cereal Chemists, St. PaulGoogle Scholar
  7. Bressani R (1995) Proceedings of the international symposium on quality protein maize EMBRAPA/CNPMS, Sete Lagaos, Brazil, pp 41–63Google Scholar
  8. Brochetto-Braga MR, Leite A, Arruda P (1992) Partial purification and characterization of lysine-ketoglutarate reductase in normal and opaque-2 maize endosperms. Plant Physiol 98:1139–1147PubMedCrossRefGoogle Scholar
  9. Carneiro NP, Hughes PA, Larkins BA (1999) The eEF1A gene family is differentially expressed in maize endosperm. Plant Mol Biol 41:801–813PubMedCrossRefGoogle Scholar
  10. Damerval C, Devienee D (1993) Quantification of dominance for proteins, pleiotropically affected by opaque-2 in maize. Heredity 70:38–51CrossRefGoogle Scholar
  11. Danson JW, Mercy M, Michael K, Martin L, Alex K, Alpha D (2006) Marker assisted introgression of opaque2 gene into herbicide resistant elite maize inbred lines. Afr J Biotech 5:2417–2422Google Scholar
  12. De Bosque C, Castellanos EJ, Bressani R (1988) Reporte annual. INCAP, GuatemalaGoogle Scholar
  13. Dhillon BS, Prasanna BM (2001) In: Chopra VL (ed) Breeding field crops, Oxford & IBH, New Delhi, pp 149–185Google Scholar
  14. Frisch M, Bohn M, Melchinger RAE (1999a) Comparison of selection strategies for marker assisted back crossing of a gene. Crop Sci 39:1295–1301CrossRefGoogle Scholar
  15. Frisch M, Bohn M, Melchinger RAE (1999b) Minimum sample size and optimum positioning of flanking markers in assisted back crossing for transfer of target gene. Crop Sci 39:967–975CrossRefGoogle Scholar
  16. Frizzi A, Huang S, Gilbertson LA, Armstrong TA, Luethy MH, Malvar TM (2008) Modifying lysine biosynthesis and catabolism in corn with a single bifunctional expression/silencing transgene cassette. Plant Biotech J 6:13–21Google Scholar
  17. Gaziola SA, Alessi ES, Guimaraes PEO, Damerval C, Azevedo RA (1999) Quality protein maize: a biochemical study of enzymes involved in metabolism. J Agri Food Chem 47:1268–1275CrossRefGoogle Scholar
  18. Gevers HO, Lake JK (1992) Development of modified opaque-2 maize in South Africa. In: Mertz ET (ed) Quality protein maize. American Association of Cereal Chemists, St. Paul, pp 111–121Google Scholar
  19. Gibbon BC, Larkins BA (2005) Molecular genetics approaches to developing quality protein maize. Trends Genet 21:227–233PubMedCrossRefGoogle Scholar
  20. Graham GG, Placko RP, Maclean WC (1980) Nutritional value of normal, opaque2 and, sugary 2 and opaque 2 maize hybrids for children and infants 2. Plasma free amino acids. J Nutr 110:1070–1074PubMedGoogle Scholar
  21. Gupta HS, Aggarwal PK, Mahajan V, Mani VP, Bisht GS, Kumar A, Verma P, Babu R (2009) Quality protein maize for nutritional security : rapid development of short duration hybrids through molecular marker assisted breeding. Curr Sci 96:230–237Google Scholar
  22. Gutierrez-Rojas A, Scott MP, Leyva OR, Menz M, Betran J (2008) Phenotypic characterization of quality protein maize endosperm modification and amino acid contents in a segregating recombinant inbred population. Crop Sci 48:1714–1722CrossRefGoogle Scholar
  23. Habben JE, Kirlies AW, Larkin BA (1993) The origin of lysine containing proteins in opaque-2 maize endosperm. Plant Mol Biol 23:825–838PubMedCrossRefGoogle Scholar
  24. Holding DR, Hunter BG, Chung T, Gibbon BC, Ford CF, Bharti AK, Messing J, Hamaker BR, Larkins BA (2008) Genetic analysis of opaque-2 modifier loci in quality protein maize. Theo App Genet 117:157–170CrossRefGoogle Scholar
  25. Hospital F, Charcosset A (1997) Marker-assisted introgression of quantitative trait loci. Genetics 147:1469–1485PubMedGoogle Scholar
  26. Houmard NM, Mainville JL, Bonin CP, Huang S, Luethy MH, Malvar TM (2007) High-lysine corn generated by endosperm-specific suppression of lysine catabolism using RNAi. Plant Biotech J 5:605–614CrossRefGoogle Scholar
  27. Huang SS, Adams WR, Zhou Q, Malloy KP, Voyles DA, Anthony J, Kriz AL, Luethy MH (2004) Improving nutritional quality of maize proteins by expressing sense and antisense zein genes. J Agri Food Chem 52:1958–1964CrossRefGoogle Scholar
  28. Huang S, Kruger DE, Frizzi A, D'Ordine RL, Florida CA, Adams WR, Brown WE, Luethy MH (2005) High-lysine corn produced by the combination of enhanced lysine biosynthesis and reduced zein accumulation. Plant Biotech J 3:555–569CrossRefGoogle Scholar
  29. Huang S, Frizzi A, Florida CA, Kruger DE, Luethy MH (2006) High lysine and high tryptophan transgenic maize resulting from the reduction of both 19- and 22-kDα-zeins. Plant Mol Biol 61:525–535PubMedCrossRefGoogle Scholar
  30. Huang S, Frizzi A, Malvar T (2008) Genetically engineered high lysine corn. ISB News Report. pp 1–3Google Scholar
  31. Ibitoye DO, Akin-Idowu PE (2010) Marker – assisted – selection (MAS): a fast track to increase genetic gain in horticultural crop breeding. Afr J of Biotech 52:8889–8895Google Scholar
  32. Jompuk P, Wongyai W, Jamptong C, Apisitvanich S (2006) Detection of quality protein maize (QPM) using simple sequence repeat (SSR) markers and analysis of tryptophan content in endosperm. Nat Sci 40:768–774Google Scholar
  33. Krivanek AF, Groote DE, Gunaratna H, Diallo NS, Friesen D (2007) Breeding and disseminating quality protein maize for Africa. Afr J Biotech 6:312–324Google Scholar
  34. Lambert RJ, Alexander DE, Dudley JW (1969) Relative performance of normal and modified protein (opaque-2) maize hybrid. Crop Sci 9:242–243CrossRefGoogle Scholar
  35. Lawton JW, Wilson CM (1987) Proteins of the kernel. In: White PJ, Johnson LA (eds) Corn chemistry and technology. American Association of Cereal Chemists, St. Paul, pp 313–354Google Scholar
  36. Lohmer S, Maddaloni M, Motto M, Dilonzo N, Hartings A, Salamini F, Thomson RD (1991) The maize regulatory locus opaque-2 encodes a DNA binding protein which activates the transcription of the B-32 gene. Embo J 10:617–624PubMedGoogle Scholar
  37. Lopes MA, Takasaki K, Bostwick DE, Helentjaris T, Larkins BA (1995) Identification of opaque-2 modifier loci in quality-protein-maize. Mol Gen Genet 247:603–613PubMedCrossRefGoogle Scholar
  38. Lutz W (2001) The end of world population growth. Nature 412:543–545PubMedCrossRefGoogle Scholar
  39. Manna R, Okello DK, Imanywoha J, Pixley K, Edema R (2005) Enhancing introgression of the opaque-2 trait into elite maize lines using simple sequence repeats. Afr Crop Sci J 13:215–226Google Scholar
  40. Mboogoi MN, Danson JW, Kimani M (2006) Using biotechnology to develop high lysine maize. Afr J of Biotech 5:693–696Google Scholar
  41. McWhirter KS (1971) A floury endosperm, high lysine locus on chromosome 10. Corn Genet Coop Newslett 45:184Google Scholar
  42. Mertz ET, Bates LS, Nelson OE (1964) Mutant gene that changes protein composition and increases lysine content of maize endosperm. Science 145:279–280PubMedCrossRefGoogle Scholar
  43. Micic-Ignjatovic D, Ristic D, Markovic K, Lazic-Jancic V, Denic M (2008) Quality protein maize – QPM. Genetika 40:205–214CrossRefGoogle Scholar
  44. Millward DJ, Rivers JP (1989) The need for indispensable amino acids: the concepts of the anabolic drive. Diabetes-Metab Rev 5:191–211PubMedCrossRefGoogle Scholar
  45. Nelson OE (1979) More precise linkage data on fl3. Maize Genet Coop Newslett 53:56Google Scholar
  46. Nelson OE (1981) The mutations opaque-9 through opaque-13. Corn Genet Coop Newslett 55:68Google Scholar
  47. Nelson OE, Mertz ET, Bates LS (1965) Second mutant gene affecting the amino acid pattern of maize endosperm proteins. Science 150:1469–1472PubMedCrossRefGoogle Scholar
  48. Paez AV, Helm JL, Zuber MS (1969) Lysine content of opaque-2 maize kernels having different phenotypes. Crop Sci 9:251–252CrossRefGoogle Scholar
  49. Prasanna B, Sarkar K (1991) Coordinate genetic regulation of maize endosperm. Maize Genetics Perspectives ICAR, pp 74–86Google Scholar
  50. Prasanna BM, Vasal SK, Kassahun B, Singh NN (2001) Quality protein maize. Curr Sci 81:1308–1319Google Scholar
  51. Salamini F, Borghi B, Lorenzon C (1970) Effect of opaque-2 gene on yield in maize. Euphytica 19:531–538CrossRefGoogle Scholar
  52. Schmidt RJ, Burr FA, Aukerman MJ, Burr B (1990) Maize regulatory gene opaque2 encodes protein with a “leucine zipper” motif that binds to zein DNA. Proc Natl Acad Sci U S A 87:46–50PubMedCrossRefGoogle Scholar
  53. Segal G, Song R, Messing J (2003) A new opaque variant of maize by a single dominant RNA interference-inducing transgene. Genetics 165:387–397PubMedGoogle Scholar
  54. Stepansky A, Less H, Angelovici R, Aharon R, Zhu X, Galili G (2006) Lysine catabolism, an effective versatile regulator of lysine level in plants. Amino Acids 30:121–125PubMedCrossRefGoogle Scholar
  55. Vasal SK (2000) The quality protein maize story. Food Nutr Bull 21:445–450Google Scholar
  56. Vasal SK, Villegas E, Bjarnason M, Gelow B, Goertz P (1980) Genetic modifiers and breeding strategies in developing hard endosperm opaque material. In: Pollmer WG, Phillips RH (eds) Improvement of quality traits of maize for grain and silage used. Mortinus Nijhoff Press, London, pp 37–73Google Scholar
  57. Visscher PM, Haley CS, Thompson R (1996) Marker assisted introgression in backcross breeding programs. Genetics 144:1923–1932PubMedGoogle Scholar
  58. Wang XL, Woo YN, Kim CS, Larkins BA (2001) Quantitative trait locus mapping of loci influencing elongation factor 1 alpha content in maize endosperm. Plant Physiol 125:1271–1282PubMedCrossRefGoogle Scholar
  59. Wilson CM, Shewry PR, Miflin BJ (1981) Maize endosperm proteins compared by sodium dodecyl gel electrophoresis and isoelectric focussing. Cereal Chem 58:275–281Google Scholar
  60. Wu RL, Lou XY, Ma CX, Wang XL, Larkins BA, Casella G (2002) An improved genetic model generates high-resolution mapping of QTL for protein quality in maize endosperm. Proc Natl Acad Sci U S A 99:11281–11286PubMedCrossRefGoogle Scholar
  61. Wu YR, Holding DR, Messing J (2010) γ-zein are essential for endosperm modification in quality protein maize. Proc Natl Acad Sci USA 29:12810–12815CrossRefGoogle Scholar
  62. Xu Y, Crouch JH (2008) Marker assisted selection in plant breeding: from publication to practice. Crop Sci 48:391–407CrossRefGoogle Scholar
  63. Yang W, Zheng Y, Zheng W, Feng R (2005) Molecular genetic mapping of high lysine mutant gene (Opaque-16) and the double recessive effect with opaque-2 in maize. Mol Breed 15:257–269CrossRefGoogle Scholar
  64. Zhang WL, Yang WP, Chen ZW, Wang MC, Yang LQ, Cai YL (2010) Molecular marker assisted selection for o2 introgression lines with o16 gene in corn. Acta Agronomica Sinica 36:1302–1309Google Scholar

Copyright information

© Springer India 2014

Authors and Affiliations

  1. 1.School of Agricultural BiotechnologyPunjab Agricultural UniversityLudhianaIndia
  2. 2.Department of Plant Breeding and GeneticsPunjab Agricultural UniversityLudhianaIndia

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