Molecular cloning and characterization of high-affinity potassium transporter (AlHKT2;1) gene promoter from halophyte Aeluropus lagopoides

HKT subfamily II functions as Na+ - K+ co-transporter and prevents plants from salinity stress. A 760 bp promoter region of AlHKT2:1 was isolated, sequenced and cloned. The full length promoter D1, has many cis-regulatory elements like MYB, MBS, W box, ABRE etc. involved in abiotic stress responses. D1 and subsequent 5' deletions were cloned into pCAMBIA1301 and studied for its efficacy in stress conditions in heterologous system. Blue colour staining was observed in flower petals, anther lobe, and dehiscence slit of anther in To plants. The T-1 seedlings showed staining in leaf veins, shoot vasculature and root except root tip. T-1 seedlings were subjected to NaCl, KCl, NaCl + KCl and ABA stresses. GUS activity was quantified by 4-methylumbelliferyl glucuronide (4-MUG) assay under control and stress conditions. The smallest deletion- D4 also showed GUS expression but highest activity was observed in D2 as compared to full length promoter and other deletions. The electrophoretic mobility shift assay using stress-induced protein with different promoter deletions revealed more prominent binding in D2. These results suggest that AlHKT2:1 promoter is involved in abiotic stress response and deletion D2 might be sufficient to drive the stress-inducible expression of various genes involved in providing stress tolerance in plants. (C) 2021 Elsevier B.V. All rights reserved.

Automated summary

The HKT subfamily II functions as a Na+ - K+ co-transporter, providing resistance to salinity stress in plants. Through cloning and studying the promoter region of AlHKT2:1, it was found that deletion D2 may be sufficient to drive stress-inducible expression of genes involved in stress tolerance in plants.