Comprehensive RNAseq analysis for identification of genes expressed
under terminal heat stress in lentil
Abstract
Lentil crop is highly sensitive to abrupt increase in temperature during
mid to late reproductive stage leading to severe reduction of biomass
and seed yield. Therefore, in the present investigation, RNAseq analysis
was carried out between IG4258 (heat tolerant) and IG3973 (heat
sensitive) lentil genotypes at reproductive stage under both normal and
heat stress conditions in the field. It resulted in a total of 2,09,549
assembled transcripts and among these 1,61,809 transcripts had coding
regions of which 94,437 transcripts were annotated. The differential
gene expression analysis showed up-regulation of 678 transcripts and
down regulation of 680 transcripts between tolerant and susceptible
genotypes at early reproductive stage. While 76 transcripts were
up-regulated and 47 transcripts were down-regulated at late reproductive
stage under heat stress conditions. The validation of 12 up- or down
transcripts through RT-PCR corresponded well with expression analysis
data of RNAseq with a correlation of R2=0.89. KEGG pathways enrichment
analysis identified transcripts associated with genes encoding
proteins/regulating factors related to different metabolic pathways
including signal transduction, fatty acid biosynthesis, rRNA processing,
ribosome biogenesis, gibberellin (GA) biosynthesis and riboflavin
biosynthesis. This analysis also identified 21,487 genic-SSRs that can
be used in molecular mapping of heat tolerant genes in lentil.