Genome specificity of primers to puroindoline genes

  • O. I. Sozinova
  • N. A. Kozub
  • I. A. Sozinov
  • Ya. B. Blume
DOI: https://doi.org/10.7124/FEEO.v22.947

Abstract

Aim. Genome specificity of primers for amplification of complete coding sequences of puroindoline genes was studied. Methods. PCR with gene-specific primers was performed for amplification of puroindoline genes of wheat and related species. Results. The primer pair designated PinaDH is not gene-specific as it yields two products of amplification of the genes Pina-1 and Gsp-1. This primers pair is not specific for the Pina-1 gene of the genome V of D. villosum. The primer pairs designated PinaCM and Pinb are gene-specific as they produce one amplification product of respective length. We have demonstrated that the PinaCM primer pair is also specific for the genomes D, E and V, and the Pinb primer pair is not specific for the genome V. Conclusions. Gene and genomic specificity of primers for puroindoline genes was refined. Primers suitable for further direct sequencing of coding parts of puroindoline genes of wheat and related species were chosen.

Keywords: puroindoline, Triticum aestivum, Aegilops, Dasypyrum, genome specificity.

References

Douliez J.P., Michon T., Elmorjani K., Marion D. Minireview: Structure, biological and technological functions of lipid transfer proteins and indolines, the major lipid binding proteins from cereal kernels. J. Cereal Sci. 2000. Vol. 32, No. 1. P. 1–20. doi: 10.1006/jcrs.2000.0315.

Dubreil L., Gaborit T., Bouchet B., Gallant D.J., Broekaert W.F., Quillien L., Marion D. Spatial and temporal distribution of the major isoforms of puroindolines (puroindoline-a and puroindoline-b) and nonspecific lipid transfer protein (ns-LTPle1 ) of Triticum aestivum seeds. Relationships with the in vitro antifungal properties. Plant Sci. 1998. Vol. 138. P. 121–135. doi: 10.1016/S0168-9452(98)00121-6.

Pauly A., Pareyt B., Fierens E., Delcour J.A. Wheat (Triticum aestivum L. and T. turgidum L. ssp. durum) kernel hardness: I. Current view on the role of puroindolines and polar lipids. Comprehensive Reviews in Food Science and Food Safety. 2013. Vol. 12. P. 413–426. doi: 10.1111/1541-4337.12018.

Morris C.F. Puroindolines: the molecular genetic basis of wheat grain hardness. Plant Mol. Biol. 2002. Vol. 48. P. 633–647. doi: 10.1023/A:1014837431178.

Gautier M.F., Aleman M.E., Guirao A., Marion D., Joudrier P. Triticum aestivum puroindolines, two basic cystine-rich seed proteins: cDNA sequence analysis and developmental gene expression. Plant Mol. Biol. 1994. Vol. 25. Р. 43–57. doi: 10.1007/BF00024197.

Elmorjani K., Geneix N., Dalgalarrondo M., Branlard G., Marion D. Wheat grain softness protein (Gsp1) is a puroindoline-like protein that displays a specific post-translational maturation and does not interact with lipids. J. Cereal Sci. 2013. Vol. 58. P. 117–122. doi: 10.1016/j.jcs.2013.03.012.

Giroux J., Morris C.F. Wheat grain hardness results from highly conserved mutations in the friabilin components puroindoline a and b. PNAS USA. 1998. Vol. 95. P. 6262–6266. doi: 10.1073/pnas.95.11.6262

Gautier M.F., Cosson P., Guirao A., Alary R., Joudrier P.Puroindoline genes are highly conserved in diploid ancestor wheats and related species but absent in tetraploid Triticum species. Plant Sci. 2000. Vol. 153, No. 1. P. 81–91. doi: 10.1016/S0168-9452(99)00258-7.

Charles M., Tang H., Belcram H., Paterson A., Gornicki P, Chalhoub B. Sixty million years of evolution of soft grain trait in grasses: emergence of the softness locus from the common ancestor of Pooideae and Ehrhartoideae, after their divergence from Panicoideae. Mol. Biol. Evol. 2009. Vol. 26, No. 7. P. 1651–1661. doi: 10.1093/molbev/msp076.

Massa A., Morris C.F. Molecular evolution of the puroindoline-a, puroindoline-b, and grain softness protein-1 genes in the tribe Triticeae. J. Mol. Evol. 2006. Vol. 63, No. 4. P. 526–536. doi: 10.1093/molbev/msp076.

Massa A.N., Morris C.F., Gill B.S. Sequence diversity of puroindoline-a, puroindoline-b, and the grain softness protein genes in Aegilops tauschii Coss. Crop. Sci. 2004. Vol. 44. P. 1808–1816. doi: 10.2135/cropsci2004.1808

Maniatis T., Fritsch E.F., Sambrook J. Molecular cloning: a laboratory manual. Moscow: Mir, 1984. 480 p.