The role of PQL genes in response to salinity tolerance in Arabidopsis thaliana and barley

Increasing salinity is a worldwide problem, but the knowledge on how salt enters the roots of plants remains largely unknown. Non-selective cation channels (NSCCs) have been suggested to be the major pathway for the entry of sodium ions (Na+) in several species. The hypothesis tested in this research is that PQ loop (PQL) proteins could form NSCCs, mediate some of the Na+ influx into the root and contribute to ion accumulation and the inhibition of growth in saline conditions. This is based on previous preliminary evidence indicating similarities in the properties of NSCC currents and currents mediated by PQL proteins, such as the inhibition of an inward cation current mediated by PQL proteins by high external calcium and pH acidification. PQL family members belonging to clade one in Arabidopsis and barley were characterized using a reverse genetics approach, electrophysiology and high-throughput phenotyping. Expression of AtPQL1a and HvPQL1 in HEK293 cells increased Na+ and K+ inward currents in whole cell membranes. However, when GFP-tagged PQL proteins were transiently overexpressed in tobacco leaf cells, the proteins appeared to localize to intracellular membrane structures. Based on q-RT-PCR, the levels of mRNA of AtPQL1a, AtPQL1b and AtPQL1c is higher in salt stressed plants compared to control plants in the shoot tissue, while the mRNA levels in the root tissue did not change in response to stress. Salt stress responses of lines with altered expression of AtPQL1a, AtPQL1b and AtPQL1c were examined using RGB and chlorophyll fluorescence imaging of plants growing in soil in a controlled environment chamber. Decreases in the levels of expression of AtPQL1a, AtPQL1b and AtPQL1c resulted in larger rosettes, when measured seven days after salt stress imposition. Interestingly, overexpression of AtPQL1a also resulted in plants having larger rosettes in salt stress conditions. Differences between the mutants and the wild-type plants were not observed at earlier stages, suggesting that PQLs might be involved in long-term responses to salt stress. These results contribute towards a better understanding of the role of PQLs in salinity tolerance and provide new targets for crop improvement.