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Overexpression of PeHKT1;1 Improves Salt Tolerance in Populus.

ABSTRACT: Soil salinization is an increasingly serious threat that limits plant growth and development. Class I transporters of the high-affinity K⁺ transporter (HKT) family have been demonstrated to be involved in salt tolerance by contributing to Na⁺ exclusion from roots and shoots. Here, we isolated the PeHKT1;1 gene from hybrid poplar based on the sequences of the Populus trichocarpa genome. The full-length PeHKT1;1 gene was 2173 bp, including a 1608 bp open reading frame (ORF) encoding 535 amino acids and containing eight distinct transmembrane domains. Multiple sequence alignment and phylogenetic analysis suggested that the PeHKT1;1 protein had a typical S⁻G⁻G⁻G signature for the P-loop domains and belonged to class I of HKT transporters. PeHKT1;1 transcripts were mainly detected in stem and root, and were remarkably induced by salt stress treatment. In further characterization of its functions, overexpression of PeHKT1;1 in Populus davidiana × Populus bolleana resulted in a better relative growth rate in phenotypic analysis, including root and plant height, and exhibited higher catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities than non-transgenic poplar under salt stress conditions. These observations indicated that PeHKT1;1 may enhance salt tolerance by improving the efficiency of antioxidant systems. Together, these data suggest that PeHKT1;1 plays an important role in response to salt stress in Populus.

SUBMITTER: Xu M

PROVIDER: S-EPMC6210203 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Mitogen-activated protein kinase (MAPK) is one of the factors of cascade reactions affecting responses to signal pathway of environmental stimuli. Throughout the life of plants, MAPK family members participate in signal transduction pathways and regulate various intracellular physiological and metabolic reactions. To gain insights into regulatory function of MAPK kinase (MAPKK) in Populus trichocarpa under salt stress, we obtained full-length cDNA of PtMAPKK4 and analyzed different expression levels of PtMAPKK4 gene in leaves, stems, and root organs. The relationship between PtMAPKK4 and salt stress was studied by detecting expression characteristics of mRNA under 150 mM NaCl stress using real-time quantitative polymerase chain reaction. The results showed that expression of PtMAPKK4 increased under salt (NaCl) stress in leaves but initially reduced and then increased in roots. Thus, salt stress failed to induce PtMAPKK4 expression in stems. PtMAPKK4 possibly participates in regulation of plant growth and metabolism, thereby improving its salt tolerance. We used Saccharomyces cerevisiae strain INVScI to verify subcellular localization of PtMAPKK4 kinase. The yeast strains containing pYES2-PtMAPKK4-GFP plasmid expressed GFP fusion proteins under the induction of d-galactose, and the products were located in nucleus. These results were consistent with network prediction and confirmed location of PtMAPKK4 enzyme in the nucleus. We tested NaCl tolerance in transgenic tobacco lines overexpressing PtMAPKK4 under the control of 35S promoter at germination stage to detect salt tolerance function of PtMAPKK4. Compared withK326 (a wild-type tobacco), lines overexpressing PtMAPKK4 showed a certain degree of improvement in tolerance, germination, and growth. NaCl inhibited growth of overexpressed line and K326 at the seedling stage. However, statistical analysis showed longer root length, higher fresh weight, and lower MDA content in transgenic lines in comparison with that in K326.

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Overexpression of PeHKT1;1 Improves Salt Tolerance in Populus.

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