Differential expression analysis of defense-related genes responsive to Tilletia indica infection in wheat

Authors: ARADHIKA TRIPATHI, RASHMI AGGARWAL, ANITA YADAV

Abstract: Karnal bunt caused by Tilletia indica is a quarantine disease responsible for qualitative and quantitative losses in wheat. The present study aimed to determine the expression of defense-related genes involved in early stages of infection in response to T. indica by using the differential display reverse transcriptase-PCR method (DDRT-PCR) in susceptible (WL 711) and resistant (HD 29) cultivars of wheat. The DDRT-PCR banding profiles generated by the random primers in combination with oligo-dT primers of resistant and susceptible wheat plants were evaluated at 0 h, 12 h, 24 h, and 48 h after inoculation. Out of 9 sets of primers, 7 sets of primers produced remarkable induced transcripts. The highest up-regulated transcripts were observed in the resistant cultivar (HD 29), whereas the down-regulated transcripts were found in the susceptible (WL 711) cultivar. Several sets of primers were designed from the defense-related gene sequences available in the GenBank database, and of these, 4 primer sets amplified the desired specific bands. Based on the putative functions, 3 defense-related genes, namely puroindoline protein PINB (AT_-_ KB2), \Beta-1,4-glucanase (AT_-_ KB3), and chitinase (AT_-_ KB7), and 1 housekeeping gene, actin (AT_-_ KB1), were characterized.

Keywords: Defense-related genes, expression, DDRT-PCR, Tilletia indica, wheat

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