Theoretical and Applied Genetics

, Volume 114, Issue 5, pp 803–814 | Cite as

Generation and characterization of low phytic acid germplasm in rice (Oryza sativa L.)

  • Qing-Long Liu
  • Xiu-Hong Xu
  • Xue-Liang Ren
  • Hao-Wei Fu
  • Dian-Xing Wu
  • Qing-Yao Shu
Original Paper


Phytic acid (PA, myo-inositol 1,2,3,4,5,6-hexakisphosphate), or its salt form, phytate, is commonly regarded as the major anti-nutritional component in cereal and legume grains. Breeding of low phytic acid (lpa) crops has recently been considered as a potential way to increase nutritional quality of crop products. In this study, eight independent lpa rice mutant lines from both indica and japonica subspecies were developed through physical and chemical mutagenesis. Among them, five are non-lethal while the other three are homozygous lethal. None of the lethal lines could produce homozygous lpa plants through seed germination and growth under field conditions, but two of them could be rescued through in vitro culture of mature embryos. The non-lethal lpa mutants had lower PA content ranging from 34 to 64% that of their corresponding parent and four of them had an unchanged total P level. All the lpa mutations were inherited in a single recessive gene model and at least four lpa mutations were identified mutually non-allelic, while the other two remain to be verified. One mutation was mapped on chromosome 2 between microsatellite locus RM3542 and RM482, falling in the same region as the previously mapped lpa1-1 locus did; another lpa mutation was mapped on chromosome 3, tightly linked to RM3199 with a genetic distance of 1.198 cM. The latter mutation was very likely to have happened to the LOC_Os03g52760, a homolog of the maize myo-inositol kinase (EC gene. The present work greatly expands the number of loci that could influence the biosynthesis of PA in rice, making rice an excellent model system for research in this area.


Oryza sativa L.  Low phytic acid (lpaMutation Gene mapping Myo-inositol kinase 


  1. Adams C, Raboy V, Krebs N, Westcott J, Lei S, Hambidge M (2001) The effect of low-phytic acid corn mutants on Zn absorption. FASEB J 73:80–85Google Scholar
  2. Andaya CB, Tai TH (2005) Fine mapping of the rice low phytic acid (lpa1) locus. Theor Appl Genet 111:489–495PubMedCrossRefGoogle Scholar
  3. Bregitzer P, Raboy V (2006) Effects of four independent low-phytate mutations on barley agronomic performance. Crop Sci 46:1318–1322CrossRefGoogle Scholar
  4. Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphorous. Anal Chem 28:1756–1758CrossRefGoogle Scholar
  5. Dorsch JA, Cook A, Young KA, Anderson JM, Bauman AT, Volkmann CJ, Murthy PPN, Raboy V (2003) Seed phosphorus and inositol phosphate phenotype of barley low phytic acid genotypes. Phytochem 62:691–706CrossRefGoogle Scholar
  6. Douglas MW, Peter CM, Boling SD, Parsons CM, Baker DH (2000) Nutritional evaluation of low phytate and high protein corns. Poult Sci 79:1586–1591PubMedGoogle Scholar
  7. Guttieri M, Bowen D, Dorsch JA, Raboy V, Souza E (2004) Identification and characterization of a low phytic acid wheat. Crop Sci 44:418–424CrossRefGoogle Scholar
  8. van Harten AM (1998) Mutation breeding—theory and practical application. Cambridge University Press, CambridgeGoogle Scholar
  9. Larson SR, Young KA, Cook A, Blake TK, Raboy V (1998) Linkage mapping of two mutations that reduce phytic acid content of barley grain. Theor Appl Genet 97:141–146CrossRefGoogle Scholar
  10. Larson SR, Rutger JN, Young KA, Raboy V (2000) Isolation and genetic mapping of a non-lethal rice (Oryza sativa L.) low phytic acid 1 mutation. Crop Sci 40:1397–1405CrossRefGoogle Scholar
  11. Li YC, Ledoux DR, Veum TL, Raboy V, Ertl DS (2000) Effects of low phytic acid corn on phosphorus utilization, performance, and bone mineralization in broiler chicks. Poult Sci 79:1444–1450PubMedGoogle Scholar
  12. Liu RH, Meng JL (2003) Mapdraw: a Microsoft Excel macro for drawing genetic linkage maps based on given genetic linkage data. Hereditas (Beijing) 25:317–321Google Scholar
  13. Lolle SJ, Victor JL, Young JM, Pruitt RE (2005) Genome-wide non-mendelian inheritance of extra-genomic information in Arabidopsis. Nature 434: 505–509PubMedCrossRefGoogle Scholar
  14. Lott JNA, Ockenden I, Raboy V, Batten GD (2000) Phytic acid and phosphorus in crops seeds and fruits: a global estimate. Seed Sci Res 10:11–33Google Scholar
  15. Loza PL, Stanton TL, Engle TE, Schutz D, Rhoads AR (2002) Effects of feeding low phytate corn varieties on growth performance, feed efficiency, serum phosphorus level, Longissimus Dorsi fatty acid composition and carcass characteristics of finishing beef cattle. J Anim Sci 80:1999–2005Google Scholar
  16. Lu YJ, Zheng KL (1992) A simple method for isolation of rice DNA. Chin J Rice Sci 6:47–48Google Scholar
  17. Meis SJ, Fehr WR, Schnebly SR (2003) Seed source effect on field emergence of soybean lines with reduced phytate and raffinose saccharides. Crop Sci 43:1336–1339CrossRefGoogle Scholar
  18. 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
  19. Mendoza C, Viteri FE, Lonnerdal B, Raboy V, Young KA, Brown KH (2001) Absorption of iron from unmodified maize and genetically altered, low-phytate maize fortified with ferrous sulfate or sodium iron EDTA. Am J Clin Nutr 73:80–85PubMedGoogle Scholar
  20. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  21. van Oijen JW, Voorrips RE (2001). JoinMapR Version 3.0 software for the calculation of genetic linkage map. Plant Res Int, WageningenGoogle Scholar
  22. Oltmans SE, Fehr WR, Welke GA, Raboy V, Peterson KL (2005) Agronomic and seed traits of soybean lines with low-phytate phosphorus. Crop Sci 45:593–598CrossRefGoogle Scholar
  23. Panaud O, Chen X, McCouch SR (1996) Development of microsatellite markers and characterization of simple sequence length polymorphism (SSLP) in rice (Oryza sativa L.). Mol Gen Genet 252:597–607PubMedGoogle Scholar
  24. Philliphy BQ, Bland JM, Evens TJ (2003) Ion chromatography of phytate in roots and tubers. J Agric Food Chem 51:350–353CrossRefGoogle Scholar
  25. Pilu R, Panzeri D, Gavazzi G, Rasmussen SK, Consonni G, Nielsen E (2003) Phenotypic, genetic and molecular characterization of a maize low phytic acid mutant (lpa 241). Theor Appl Genet 107:980–987PubMedCrossRefGoogle Scholar
  26. Pilu R, Landoni M, Cassani E, Doria E, Nielsen E (2005) The maize lpa241 mutation causes a remarkable variability of expression and some pleiotropic effects. Crop Sci 45:2096–2105CrossRefGoogle Scholar
  27. 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, Dordrecht, pp 441–477Google Scholar
  28. Raboy V (2001) Seeds for a better future: ‘low phytate’ grains help to overcome malnutrition and reduce pollution. Trends Plant Sci 6:458–462PubMedCrossRefGoogle Scholar
  29. Raboy V, Dickison DB, Below FE (1984) Variation in seed total phosphorous, phytic acid, zinc, calcium, magnesium, and protein among lines of Glycine max and G. soja. Crop Sci 24:431–434CrossRefGoogle Scholar
  30. 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 124:355–368PubMedCrossRefGoogle Scholar
  31. Rasmussen SK, Hatzak F (1998) Identification of two low-phytate barley (Hordeum vulgare L.) grain mutants by TLC and genetic analysis. Hereditas 129:107–112CrossRefGoogle Scholar
  32. Rassoulzadegan M, Grandjean V, Gounon P, Vicent S, Gillot I, Cuzin F (2006) RNA-mediated non-mendelian inheritance of an epigenetic change in the mouse. Nature 441: 469–474PubMedCrossRefGoogle Scholar
  33. Rutger JN, Raboy V, Moldenhauer KAK, Bryant RJ, Lee FN, Gibbons JW (2004) Registration of KBNT lpa1–1 low phytic acid germplasm of rice. Crop Sci 44:363CrossRefGoogle Scholar
  34. Shen SQ, Wu DX, Xia YW, Shu QY (2003) Obtaining pure insect resistant Bt lines through anther culture in rice. J Agric Biotechnol 11:561–565Google Scholar
  35. Shi JR, Wang HY, Wu YS, 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 131:507–515PubMedCrossRefGoogle Scholar
  36. Shi JR, Wang HY, Hazebroek J, Ertl DS, Harp T (2005) The maize low-phytic acid 3 encodes a myo-inositol kinase that plays a role in phytic acid biosynthesis in developing seeds. Plant J 42:408–419CrossRefGoogle Scholar
  37. Shi JR, Ertl DS, Wang HY, Li BL, Faller M, Schellin K (2006) Maize multidrug resistance-associated protein polynucleotides and methods of use. US Patent Application 20060143728Google Scholar
  38. Shukla S, Vantoai TT, Pratt RC (2004) Expression and nucleotide sequence of an INS(3)P1 synthase gene associated with low-phytate kernels in maize (Zea mays L.). J Agric Food Chem 52:465–4570Google Scholar
  39. Spencer JD, Allee GL, Sauber TE (2000a) Phosphorus bioavailability and digestibility of normal and genetically modified low-phytate corn for pigs. J Anim Sci 78:675–681Google Scholar
  40. Spencer JD, Allee GL, Sauber TE (2000b) Growing-finishing performance and carcass characteristics of pigs fed normal and genetically modified low-phytate corn. J Anim Sci 78:1529–1536Google Scholar
  41. Stevenson-Paulik J, Bastidas RJ, Chiou AT, Frye RA, York JD (2005) Generation of phytate-free seeds in Arabidopsis through disruption of inositol polyphosphate kinases. PNAS USA 102:12612–12617PubMedCrossRefGoogle Scholar
  42. Temnykh S, Park WD, Ayers N, Cartinhour S, Hauck N, Lipovich L, Cho YG, Shii T, McCouch SR (2000) Mapping and genome organization of microsatellite sequences in rice (Oryza sativa L.). Theor Appl Genet 100:697–712CrossRefGoogle Scholar
  43. Temnykh S, DeClerck G, Lukasshova A, Lipovich L, Cartinhour S, McCouch SR (2001) Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): frequency, length variation, transposon associations, and genetic marker potential. Genome Res 11:1441–1452PubMedCrossRefGoogle Scholar
  44. Veum TL, Ledoux DR, Raboy V, Ertl DS (2001) Low-phytic acid corn improves nutrient utilization of growing pigs. J Anim Sci 79:2873–2880PubMedGoogle Scholar
  45. Walker DR, Scaboo AM, Pantalone VR, Wilcox JR, Boerma HR (2006) Genetic mapping of loci associated with seed phytic acid content in CX1834-1-2 soybean. Crop Sci 46:390–397CrossRefGoogle Scholar
  46. Wilcox JR, Premachandra GS, Young KA, Raboy V (2000) Isolation of high inorganic P, low-phytate soybean mutants. Crop Sci 40:1601–1605CrossRefGoogle Scholar
  47. Xia HJ, Yang G (2005) Inositol 1,4,5-trisphosphate 3 kinases: functions and regulation. Cell Res 15:83–91PubMedCrossRefGoogle Scholar
  48. Yan F, Kersey JH, Fritts CA, Waldroup PW, Stilborn HL, Crum RC, Rice DW, Raboy V (2000) Evaluation of normal yellow dent corn and high available phosphorus corn in combination with reduced dietary phosphorus and phytase supplementation for broilers grown to market weights in litter pens. Poult Sci 79:1282–1289PubMedGoogle Scholar
  49. Yan F, Fritts CA, Waldroup PW, Stilborn HL, Rice D, Crum RC Jr, Raboy V (2003) Comparison of normal and high available phosphorus corn with and without phytase supplementation in diets for male large white turkeys grown to market weights. Int J Poult Sci 2:83–90CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Qing-Long Liu
    • 1
    • 2
  • Xiu-Hong Xu
    • 1
  • Xue-Liang Ren
    • 1
  • Hao-Wei Fu
    • 1
    • 3
  • Dian-Xing Wu
    • 1
  • Qing-Yao Shu
    • 1
    • 4
  1. 1.IAEA-Zhejiang University Collaborating Center, Institute of Nuclear Agricultural SciencesZhejiang UniversityHangzhouChina
  2. 2.Institute of Crop Science and Nuclear Technology UtilizationZhejiang Academy of Agricultural SciencesHangzhouChina
  3. 3.Jiaxing Academy of Agricultural SciencesJiaxingChina
  4. 4.Joint FAO/IAEA Division of Nuclear Techniques in Food and AgricultureInternational Atomic Energy AgencyViennaAustria

Personalised recommendations