PJB-2019-397
DEPLOYMENT OF SSR MARKERS FOR TESTING UNIFORMITY/PURITY OF A LARGE WHEAT (TRITICUM AESTIVUM L.) TILLING POPULATION
MOMINA HUSSAIN
Abstract
Aim of the present investigation was to conduct DNA-based genotyping assay, specifically exploring microsatellite regions, on wheat M2 population for identifying the genuine mutant plants (other than the natural variants) before conducting TILLING experiments. Seed of a wheat variety ‘NN-Gandum-1’ was exposed to two parameters including exposure time and temperatures. It was established that 0.8% EMS when exposed for 2 hour at 35°C was found appropriate for developing a suitable TILLING population. For determining the quality of the mutant population, a purity analysis was carried out using SSR primer pairs with high PIC values. Initially, 77 SSRs were surveyed on wheat genotypes. In total, 20 SSRs showing high PIC values were selected for surveying 3634 M2 plants. Out of these, three SSR primer pairs WMS-46, WMS-249 and WMS-311 detected natural variants in M2 TILLING population with 5.09, 7.23 and 8.29%, respectively. The While, WMS-249 and WMS-311 were located in densely mapped regions (near the centromere) than that of the WMS-46. It was concluded that 20 SSRs with high PIC value are sufficient to identify true-to-type mutants (other than natural variants) in a wheat TILLING population. This study has implications for identifying genes and their function as well as in genetic improvement of wheat. This information can also be translated on other crops.
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