The RNA-seq transcriptomic analysis reveals genes mediating salt tolerance through rapid triggering of ion transporters in a mutant barley
Abstract Regarding the complexity of the mechanisms of salinity tolerance, the use of isogenic lines or mutants that have the same genetic background but show different tolerance to salinity is a suitable method to reduce the analytical complexity to study these mechanisms. In the current study, whole transcriptome analysis was evaluated using RNA-seq method between a salt-tolerant mutant line “73-M4-30” and its wild-type “Zarjou” cultivar at a seedling stage after six hours of exposure to salt stress (300 mM NaCl). Transcriptome sequencing yielded 20 million reads for each genotype. A total number of 7116 transcripts with differential expression were identified, 1586 and 1479 of which were obtained with significantly increased expression in the mutant and the wild-type, respectively. In addition, the families of WRKY, ERF, AP2/EREBP, NAC, CTR/DRE, AP2/ERF, MAD, MIKC, HSF, and bZIP were identified as the important transcription factors with specific expression in the mutant genotype. The RNA-seq results were confirmed in several time points using qRT-PCR of some important salt-responsive genes. In general, the results revealed that the mutant compared to its wild-type via fast stomach closure and consequently transpiration reduction under the salt stress, saved more sodium ion in the root and decreased its transfer to the shoot, and increased the amount of potassium ion leading to the maintenance a high ratio [K+]/[Na+] in the shoot. Moreover, it caused a reduction in photosynthesis and respiration, resulting in the use of the stored energy and the carbon for maintaining the plant tissues, which is a mechanism of salt tolerance in plants. Up-regulation of catalase, peroxidase, and ascorbate peroxidase genes, which was probably due to the more accumulation of H2O2 in the wild-type compared to the mutant. Therefore, the wild-type initiated rapid ROS signals lead to less oxidative scavenging than the mutant. The mutant increased expression in the ion transporters and the channels related to the salinity to retain the ion homeostasis. Totally, the results demonstrated that the mutant responded better to the salt stress under both the osmotic and the ionic stress phases. Less damage was observed in the mutant compared to its wild-type under the salt stress.