Phenotypic, genetic and molecular characterization of a maize low phytic acid mutant (lpa241)
- 609 Downloads
- 62 Citations
Abstract
Phytic acid, myo-inositol 1,2,3,4,5,6-hexakisphosphate, is the major storage compound of phosphorous (P) in plants, predominantly accumulating in seeds (up to 4–5% of dry weight) and pollen. In cereals, phytic acid is deposited in embryo and aleurone grain tissues as a mixed "phytate" salt of potassium and magnesium, although phytates contain other mineral cations such as iron and zinc. During germination, phytates are broken down by the action of phytases, releasing their P, minerals and myo-inositol which become available to the growing seedling. Phytic acid represents an anti-nutritional factor for animals, and isolation of maize low phytic acid (lpa) mutants provides a novel approach to study its biochemical pathway and to tackle the nutritional problems associated with it. Following chemical mutagenesis of pollen, we have isolated a viable recessive mutant named lpa 241 showing about 90% reduction of phytic acid and about a tenfold increase in seed-free phosphate content. Although germination rate was decreased by about 30% compared to wild-type, developement of mutant plants was apparentely unaffected. The results of the genetic, biochemical and molecular characterization experiments carried out by SSR mapping, MDD-HPLC and RT-PCR are consistent with a mutation affecting the MIPS1S gene, coding for the first enzyme of the phytic acid biosynthetic pathway.
Keywords.
Maize Mutagenesis Phytic acid Myo-inositol(3) P1 synthase RACENotes
Acknowledgements
This work was supported by the Ministero dell'Universita e della Ricerca Scientifica e Tecnologica-Cofin 1998 (to G.G.). We wish to thank Mikael Blom Sorensen (Risoe National Laboratory, DK) for skilful assistance in MDD-HPLC experiments, and Dr. Anne Marie Lescure (Laboratoire de Biologie des Semences, INRA INA-PG, Versailles, F) for kindly providing phyS11 cDNA clones.
References
- Adams C, Raboy V, Krebs N, Westcott J, Lei S, Hambidge M (2000) The effect of low-phytic acid corn mutants on zinc absorption. FASEB J 14:A359.11Google Scholar
- Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleid Acids Res 25:3389–3402CrossRefGoogle Scholar
- Beckett JB (1978) B-A translocations in maize. I. Use in locating genes by the chromosome arm. J Hered 69:27–36Google Scholar
- Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756–1758Google Scholar
- Cromwell GL, Coffey RD (1991) Phosphorus – a key essential nutrient, yet a possible major pollutant – its central role in animal nutrition. In: Lyons TP (ed) Biotechnology in the feed industry. Alltech Tech Publishers, Nicholasville, Kentucky, pp 133–145Google Scholar
- Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21Google Scholar
- Ertl DS, Young KA, Raboy V (1998) Plant genetic approaches to phosphorus management in agricultural production. J Environ Qual 27:299–304Google Scholar
- Feinberg AP, Volgelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13PubMedGoogle Scholar
- Frohman MA (1990) RACE, rapid amplification of cDNAs ends, in PCR protocols: a guide to methods and applications. In: Innis MA, Gelfand DH, Sninsky J, White TJ (eds) Academic Press, San Diego, pp 28–38Google Scholar
- Hatzack F, Hubel F, Zhang W, Hansen PE, Rasmussen SK (2001) Inositol phosphates from barley low-phytate grain mutants analyzed by metal-dye detection HPLC and NMR. Biochem J 354:473–480CrossRefPubMedGoogle Scholar
- Labourè AM, Gagnon J, Leseure AM (1993) Purification and characterization of a phytase (myo-inositol-hexakisphosphate phosphohydrolase) accumulated in maize (Zea mays) seedling during germination. Biochem J 295:413–419PubMedGoogle Scholar
- Lander ES, Green P, Abrahamson P, Barlow J, Daly A, Lincoln SE, Newburg L (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181PubMedGoogle Scholar
- Larson SR, Raboy V (1999) Linkage mapping of maize and barley myo-inositol 1-phosphate synthase DNA sequences: correspondence with a low phytic acid mutation. Theor Appl Genet 99:27–36Google Scholar
- Larson SR, Young KA, Cook A, Blake TK, Raboy V (1998) Linkage mapping of two mutations that reduce phytic acid contents of barley grain. Theor Appl Genet 97:141–146CrossRefGoogle Scholar
- Larson SR, Rutger JN, Yung KA, Raboy V (2000) Isolation and genetic mapping of a non-lethal rice (Oryza sativa L.) low phytic acid mutation. Crop Sci 40:1397–1405Google Scholar
- Loewus FA, Murthy PPN (2000) myo-Inositol metabolism in plants. Plant Sci 150:1–19Google Scholar
- Lott JNA (1984) Accumulation of seed reserves of phosphorous and other minerals. In: Murray DR (ed) Seed physiology, vol 1. Academic Press, Sydney, Australia, pp 139–166Google Scholar
- Lott JNA, Ockenden I, Raboy V, Batten GD (2000) Phytic acid and phosphorus in crop seeds and fruits: a global estimate. Seed Sci Res 10:11–33Google Scholar
- Mayr GW (1988) A novel metal-dye detection system permits picomolar-range h.p.l.c. analysis of inositol polyphosphates from non-radioactively labeled cell or tissue specimens. Biochem J 254:585–591PubMedGoogle Scholar
- Mendoza C, Viteri FE, Lonnerdal B, Young KA, Raboy V, Brown KH (1998) Effect of genetically modified, low-phytic acid maize on absorption of iron from tortillas. Am J Clin Nutr 68:1123–1128PubMedGoogle Scholar
- Neuffer MG, Coe EH (1978) Paraffin oil technique for treating pollen with chemical mutagens. Maydica 23:21–28Google Scholar
- O'Dell BL, de Boland AR, Koirtyohann SR (1972) Distribution of phytate and nutritionally important elements among the morphological components of cereal grains. J Agric Food Chem 20:718–721Google Scholar
- Pilu R, Gavazzi G, Nielsen E (2002) Isolation and preliminary characterization of a maize low phytic acid mutant. Maize Genet Coop Newslett 76:46Google Scholar
- Raboy V (1990) The biochemistry and genetics of phytic acid synthesis. In: Morre DJ, Boss W, Loewus FA (eds) Inositol metabolism in plants. Alan R. Liss, New York, pp 52–73Google Scholar
- Raboy V (1997) Accumulation and storage of phosphate and minerals. In: Larkins BA, Vasil IK (eds) Cellular and molecular biology of plant seed development. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 441–447Google Scholar
- Raboy V, Gerbasi PF (1996) Genetics of myo-inositol phosphate synthesis and accumulation. Subcellular Biochemistry: myoinositol phosphates, phosphoinositides, and signal transduction. In: Biswas BB, Biswas S (eds) Plenum Press, New York, pp 257–258Google Scholar
- Raboy V, Dickinson DB (1987) The timing and rate of phytic acid accumulation in developing soybean seeds. Plant Physiol 85:841–844Google Scholar
- Raboy V, Gerbasi PF, Young KA, Stoneberg SD, Pickett SG, Bauman AT, Murthy PPN, Sheridan WF, Ertl DS (2000) Origin and seed phenotype of maize low phytic acid 1-1 and low phytic acid 2-1. Plant Physiol 124:355–368CrossRefPubMedGoogle Scholar
- Raboy V, Young KA, Dorsch JA, Cook A (2001) Genetics and breeding of seed phosphorus and phytic acid. J Plant Physiol 158:489–497Google Scholar
- Rasmussen SK, Hatzack F (1998) Identification of two low-phytate barley (Hordeum vulgare L.) grain mutants by TLC analysis. Hereditas 129:107–112Google Scholar
- Sebastian SA, Kerr PS, Pearlstein RW, Hitz WD (2000) Soybean germplasm with novel genes for improved digestibility. In: Drackley JK (ed) Soy in animal nutrition. Federation of Animal Science Societies, Savoy, Illinois, pp 56–74Google Scholar
- Shi J, Wang H, Wu Y, Hazebroek J, Meeley RB, Ertl DS (2003) The maize low phytic acid mutant lpa2 is caused by mutation in an Inositol phosphate kinase gene. Plant Physiol 131:507–515CrossRefPubMedGoogle Scholar
- Spencer JD, Allee GL, Sauber TE (2000) Phosphorus bioavailability and digestibility of normal and genetically modified low-phytate corn for pigs. J Anim Sci 78:675–681PubMedGoogle Scholar
- Sugiura SH, Raboy V, Young KA, Dong FM, Hardy RW (1998) Availability of phosphorus and trace elements in low phytate varieties of barley and corn for rainbow trout (Oncorhynchus mykiss). Aquaculture 170:285–286CrossRefGoogle Scholar
- Tunen van AJ, Koes RE, Spelt CE, van der Kroll AR, Stuitje AR, Mol JNM (1988) Cloning of two chalcone flavanone isomerase genes from Petunia hybrida: coordinate, light-regulated and differential expression of flavonoid genes. EMBO J 14:2350–2363Google Scholar
- Wright AD, Moehlenkamp CA, Perrot GH, Neuffer MG, Cone KC (1992) The maize auxtrophic mutant orange pericarp is defective in duplicate genes for tryptophane synthase. Plant Cell 4:711–719CrossRefPubMedGoogle Scholar
- Yoshida KT, Wada Y, Koyama H, Mizobuki-Fukuoka R, Naito S (1999) Temporal and spatial patterns of accumulation of the transcript of myo-inositol-1-phosphate synthase and phytic acid-containing particles during seed development in rice. Plant Physiol 119:65–72CrossRefPubMedGoogle Scholar