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

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.