Identification of differentially expressed genes in salt-tolerant oilseed sunflower (Helianthus annuus L.) genotype by RNA sequencing.

Abstract

Sunflower (Helianthus annuus L.) is widely planted as an oilseed crop worldwide. Salt stress is one of the major abiotic stresses that negatively affect crop growth and productivity. To counter the negative impact of salt stress, plants have developed avoidance and tolerance mechanisms. Developing salt-tolerant genotypes requires understanding the molecular basis of adaptive mechanisms in depth. Although using model plants i.e., Arabidopsis has improved our understanding of salt tolerant mechanisms, the relative impotence and regulation mechanisms vary among plant species due to differences in genetic and metabolic backgrounds. On the other hand, sunflower is a highly polymorphic plant due to its cross-pollinated behavior which provides different salt-tolerant genotypes available for comparative analyses. In order to gain a better view of molecular mechanisms involved in salt tolerance in sunflower, RNA sequencing analysis was realized by evaluating a tolerant genotype (AS5305) with two biological replicates under control and salt stress conditions in a controlled environment. Salinity stress was applied from NaCl resource at the 8-leaf stage and samplings were done at 24h post salt stress application. Sequencing data were analyzed using tuxedo software suite. Blast2GO software and the KEGG database were used to identify the functional tasks of each of the assembled transcripts. Analysis of genes with robust expression (i.e., with FPKM > 1 in at least one sample) revealed a total of 121 significantly expressed genes between the saline-stressed and control samples. The differential expression of 11 genes was confirmed by real-time PCR. In the following, the cDNA of MYB44 as one of the selected candidate genes involved in salt tolerance was isolated, cloned, and sequenced for comparison. Overall, the results of the current study may pave the way for the accurate selection of genes involved in salinity to be used in molecular-genetics-assisted breeding programs. In addition, making use of the identified genes may help relieve the damages arising from the salt stress in sunflowers.