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Identification of opaque-2 genotypes in segregating populations of Quality Protein Maize by analysis of restriction fragment length polymorphisms

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Abstract

Quality Protein Maize (QPM) is a name given to genetically modified opaque-2 maize with hard endosperm. The opaque-2 mutation conditions a reduction in the amount of zein seed storage protein; zeins are deficient in the essential amino acids lysine and tryptophan, and mutant seed have a higher nutritional value. To utilize the potential of opaque-2 maize, elite inbreds can be converted to o2/o2 forms and subsequently to hard endosperm opaque-2. Since opaque-2 is recessive and endosperm specific, conventional backcross procedures to convert elite inbreds to opaque-2 forms are inefficient. To alleviate this problem, a marker-assisted selection procedure was developed for the Texas A&M University Quality Protein Maize breeding program. Hybridization of an O2 cDNA probe to blots of DNA from plants carrying O2 and o2 alleles showed that restriction fragment length polymorphisms (RFLPs) exist between the W64A o2 allele and O2 alleles of Mo17 and TX5855 inbred lines. To identify the opaque2 genotypes in segregating populations, an RFLP marker assay combining the O2 cDNA probe and HindIII-digestion of genomic DNA was developed. The effectiveness of the O2 RFLP marker assay was tested under field conditions using F2 and backcross populations of several hard endosperm opaque-2 lines. A comparison of the genotypes identified by RFLP analysis with the seed phenotypes of the next generation indicated that this procedure is accurate and can be used for identifying O2/O2, O2/o2, and o2/o2 genotypes of individual juvenile plants in breeding populations.

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References

  • Botstein D, White R, Skolnick M, Davis RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32:314–331

    CAS  PubMed  Google Scholar 

  • Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1996

    CAS  PubMed  Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    CAS  PubMed  Google Scholar 

  • 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, Minn., pp 79–97

    Google Scholar 

  • Glover DV (1988) Corn protein and starch-genetics and breeding and value in foods and feeds. In: Wilkinson D (ed) 43rd Annu Corn Sorghum Res Conf. American Seed Trade Assoc. Washington, DC., pp 106–130

    Google Scholar 

  • Glover DV, Mertz ET (1987) Corn. In: Olson RA, Frey KJ (eds) Nutritional quality of cereal grains: genetic and agronomic improvement. Am Soc Agron, Madison, Wis., pp 183–336

    Google Scholar 

  • Helentjaris T, King G, Slocum M, Siedenstrang C, Wegman S (1985) Restriction fragment length polymorphisms as probes for plant diversity and their development as tools for applied plant breeding. Plant Mol Biol 5:109–118

    Google Scholar 

  • Kodrzycki R, Boston RS, Larkins BA (1989) The opaque-2 mutation of maize differentially reduces zein gene transcription. Plant Cell 1:105–114

    Google Scholar 

  • Larkins BA, Lopes MA, Lending CR (1992) A genetic, biochemical and ultra-structural analysis of modified opaque-2 maize. In: Mertz ET (ed) Quality protein maize. American Association of Cereal Chemists, St. Paul, Minn., pp 174–204

    Google Scholar 

  • Maddaloni M, Di Fonzo D, Hartings H, Lazzaroni N, Salamini F, Thompson R, Motto M (1989) The sequence of the zein regulatory gene opaque-2 (O2) of Zea mays. Nucleic Acids Res 17:7532

    Google Scholar 

  • Mertz ET, Bates LS, Nelson OE (1964) Mutant gene that changes protein composition and increases lysine content of maize endosperm. Science 145:279–180

    Google Scholar 

  • Mullis KB, Faloona FA (1987) Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Method Enzymol 155:335–350

    Google Scholar 

  • Mullis KB, Faloona FA, Scharf S, Saiki R, Horn G, Erlich H (1986) Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harbor Symp Quant Biol 51:263–273

    Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight DNA. Nucleic Acids Res 8:4321

    CAS  PubMed  Google Scholar 

  • Murray MG, Yu Ma, Severson JR, West DP, Cramer JH (1988) Restriction fragment length polymorphisms: what are they and how can breeders use them. In: Wilkinson D (ed) 43rd Annu Corn Sorghum Res Conf. American Seed Trade Assoc, Washington, D.C., pp 73–87

    Google Scholar 

  • Paulis J W, Bietz JA, Felker FC, Nelson TC (1992) Evaluating quality protein maize genotypes by reversed-phase high-performance liquid chromatography. In: Mertz ET (ed) Quality protein maize. American Association of Cereal Chemists, St.Paul, Minn., pp 122–140

    Google Scholar 

  • Reed KC, Mann DA (1985) Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res 13:7202–7221

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y.

    Google Scholar 

  • Schmidt RJ, Burr FA, Aukerman MJ, Burr B (1990) Maize regulatory gene opaque-2 encodes a protein with a “leucine-zipper” motif that binds to zein DNA. In: Proc Natl Acad Sci USA 87:46–50

    Google Scholar 

  • Soave C, Di Fonzo N, Manzocchi L, Salamini F (1986) Purification and properties of an endospermic protein of maize associated with the opaque-2 and opaque-6 genes. Planta 167:587–594

    Google Scholar 

  • Stuber CW (1992) Biochemical/molecular markers in plant breeding. In: Janick J (ed) Plant breeding reviews, vol 9. John Wiley & Son, New York, pp 37–57

    Google Scholar 

  • Tanksley SD, Young ND, Patterson AH, Bonierbale MW (1989) RFLP mapping in plant breeding: new tools for an old science. Biotechnology 7:257–264

    CAS  Google Scholar 

  • Taylor B H, Manhart JR, Amasino RM (1993) Isolation and characterization of Plant DNA. In: Glick BR, Thompson JE (eds) Methods in plant molecular biology and biotechnology, CRC Press, Baton Rouge

    Google Scholar 

  • Tsai CY (1983) Genetics of storage protein in maize. In: Janick J (ed) Plant breeding reviews, vol1. AVI Publi Co, Westport, Conn., pp 103–128

    Google Scholar 

  • Villegas E, Vasal SK, Bjarnason M (1992) Quality protein maize — what is it and how it was developed? In: Mertz ET (ed) Quality protein maize. American Association of Cereal Chemists, St. Paul, Minn., pp 49–78

    Google Scholar 

  • Walton M, Helentjaris J (1987) Application of restriction fragment length polymorphism (RFLP) technology to maize breeding. In: Wilkinson D (ed) 42nd Annu Corn Sorghum Res Conf. American Seed Trade Assoc, Washington, D.C., pp 48–75

    Google Scholar 

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Communicated by A. L. Kahler

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Kata, S.R., Taylor, B.H., Bockholt, A.J. et al. Identification of opaque-2 genotypes in segregating populations of Quality Protein Maize by analysis of restriction fragment length polymorphisms. Theoret. Appl. Genetics 89, 407–412 (1994). https://doi.org/10.1007/BF00225374

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