Tagging resistance genes in kenaf to drought stress

Abstract

The present work was conducted to identify genes responsible for drought tolerance among ten kenaf genotypes. These genotypes were exposed to drought stress and subsequently subjected to molecular analysis using RNA sequencing. Drought stress was induced by altering watering intervals as follows: WT0 (watered every two days or as required), WT4 (every four days), WT6 (every six days), WT8 (every eight days), and WT10 (every ten days). RNA analysis was performed on the younger leaves after stress induction. Ten reference genes (18s rRNA, ACT4, EF1α, MZA, PP2A, PTB, RAN, TUBα, UBC, and UBQ) were used to develop primers (forward and reverse). RNA was extracted from the plants using the Quick-RNA (Plant Mini-prep kit) manufactured by Zymo Research Corporation. The RNA was then converted to cDNA using a cDNA kit, followed by purity assessment via agarose gel electrophoresis. The purified DNA was amplified using PCR. The PCR products were sequenced, and the sequences were analyzed using BLAST in NCBI. Results showed the presence of the 18s rRNA gene in both Ifeken 400 and Ifeken 100. The study suggests that 18s rRNA is associated with drought-tolerant traits observed in these two genotypes, which could serve as a useful tool in future crop breeding programs.