Journal of Crop Science and Biotechnology

, Volume 21, Issue 1, pp 83–88 | Cite as

Identification of PCR-based DNA Marker Linked to High Phytase Level of Wheat

  • Amit Vashishth
  • Sewa Ram
  • Vikas Beniwal
Research Article


The phytase is a key enzyme to hydrolyze phytic acid present in wheat grains and improves the bio-availability of micronutrients in monogastric animals. Phytase trait being contributed by specific regions of the genome requires identification of these regions, using suitable molecular markers. Hence, in the present investigation we attempted to develop a PCR-based marker that detects the phytase level in wheat. Six sets of PCR primers were designed on the basis of nucleotides sequence variation found in the sequence of both varieties. Out of six set of primers, one set amplified two different sized bands, i.e. 334 bp and 295 in two wheat cultivars C-306 (low phytase) and DBW 17 (high phytase), respectively. It exhibited a polymorphic banding pattern with length polymorphism and clearly separating low and high phytase genotypes. The primer set was also used for PCR of 46 synthetic hexaploids and 46 release varieties of wheat to validate the developed markers. Association among identified markers and phytase activity was found to be at 99.9% confidence level based on Fisher’s exact test (F-test). Therefore, this PCR primer set will be useful to select the wheat germplasm having high phytase levels and also in wheat breeding programs aimed at improving phytase levels in bread wheat cultivars.

Key words

Phytase phytic acid micronutrients polymorphism wheat 


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  1. Bohn L, Meyer AS, Rasmussen SK. 2008. Phytate: impact on environment and human nutrition-A challenge for molecular breeding. J. Zhejiang Univ. Sci. B 9: 165–191CrossRefPubMedPubMedCentralGoogle Scholar
  2. Brinch-Pedersen H, Sorensen LD, Holm PB. 2002. Engineering crop plants: getting a handle on phosphate. Trends Plant Sci. 7: 118–125CrossRefPubMedGoogle Scholar
  3. Dionisio G, Madsen CK, Holm PB, Welinder KG, Jorgensen M, Stoger E, Arcalis E, Brinch-Pedersen H. 2011. Cloning and characterization of purple acid phosphatase phytases from wheat, barley, maize, and rice. Plant Physiol. 156: 1087–1100CrossRefPubMedPubMedCentralGoogle Scholar
  4. Ertl DS, Young KA, Raboy V. 1998. Plant genetic approaches to phosphorus management in agricultural production. J. Environ. Qual. 27: 299–304CrossRefGoogle Scholar
  5. He XY, He ZH, Zhang LP, Sun DJ, Morris CF, Fuerst EP. 2007. Allelic variation of polyphenol oxidase (PPO) genes located on chromosomes 2A and 2D and development of functional markers for the PPO genes in common wheat. Theor. Appl. Genet. 115:47–58 doi:10.1007/s00122-007-0539-8CrossRefPubMedGoogle Scholar
  6. Kumar SS, Tamilkumar P, Senthil N, Nagarajan P, Thangavelu AU, Raveendran M, Vellaikumar S, Ganesan KN, Balagopal R, Vijayalakshmi G, Shobana V. 2014. Marker-assisted selection of low phytic acid trait in maize (Zeamays L.). Hereditas 151: 20–27CrossRefGoogle Scholar
  7. Mendoza C, Viteri FE, Lonnerdal B. 1998. Effect of genetically modifi ed, low-phytic acid maize on absorption of iron from tortillas Am. J. Clin. Nutr. 68: 1123–1128CrossRefGoogle Scholar
  8. Oliver RE, Yang C, Hu G, Raboy V, Zhang M. 2009. Identification of PCR-based DNA markers flanking three low phytic acid mutant loci in barley. J. Plant Breed.Crop Sci. 1: 087–093Google Scholar
  9. Orita M, Iwahana H, Kanazawat H, Hayashi K, Sekiya T. 1989. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc. Nat. Acad. Sci. 86: 2766–2770CrossRefPubMedPubMedCentralGoogle Scholar
  10. Naidoo R, Watson GMF, Tongoona P, Derera J, Laing MD. 2013. Development of a single nucleotide polymorphism (SNP) marker for detection of the low phytic acid (lpa1-1) gene used during maize breeding. Afr. J. Biotechnol. 12: 892–900Google Scholar
  11. Rozen S, Skaletsky HJ. 2000. Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: Meth. Mol. Biol., Humana Press, Totowa, NJ, 132: 365–386Google Scholar
  12. Tommasini L, Yahiaoui N, Srichumpa P, Keller B. 2006. Development of functional markers specific for seven Pm3 resistance alleles and their validation in the bread wheat gene pool. Theor. Appl. Genet. 114: 165–175, doi:10.1007/s00122-006-0420-1CrossRefPubMedGoogle Scholar
  13. Verma A, Ram S, Dalal S. 2013. Characterization of a phytase (TaPAPhy_a 1.1) gene in an Indian wheat cultivar. Cereal Res. Commun. 42: 1–9Google Scholar
  14. Zhao XL, Ma W, Gale KR, Lei ZS, He ZH, Sun QX. 2007. Identification of SNPs and development of functional markers for LMW-GS genes at Glu-D3 and Glu-B3 loci in bread wheat (Triticum aestivum L.). Mol. Breed. 20: 223–231, doi:10.1007/s11032-007-9085-yCrossRefGoogle Scholar

Copyright information

© Korean Society of Crop Science and Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Quality and Basic Sciences LaboratoryICAR-IIWBRKarnalIndia
  2. 2.Department of BiotechnologyMMUMullana, AmbalaIndia

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