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HKT1;5 Transporter Gene Expression and Association of Amino Acid Substitutions With Salt Tolerance Across Rice Genotypes.
Saturday, 2019/12/14 | 09:07:03

Shohan MUSSinha SNabila FHDastidar SGSeraj ZI.

Front Plant Sci. 2019 Nov 4;10:1420. doi: 10.3389/fpls.2019.01420. eCollection 2019.

Abstract

Plants need to maintain a low Na+/K+ ratio for their survival and growth when there is high sodium concentration in soil. Under these circumstances, the high affinity K+ transporter (HKT) and its homologs are known to perform a critical role with HKT1;5 as a major player in maintaining Na+ concentration. Preferential expression of HKT1;5 in roots compared to shoots was observed in rice and rice-like genotypes from real time PCR, microarray, and RNAseq experiments and data. Its expression trend was generally higher under increasing salt stress in sensitive IR29, tolerant Pokkali, both glycophytes; as well as the distant wild rice halophyte, Porteresia coarctata, indicative of its importance during salt stress. These results were supported by a low Na+/K+ ratio in Pokkali, but a much lower one in P. coarctata. HKT1;5 has functional variability among salt sensitive and tolerant varieties and multiple sequence alignment of sequences of HKT1;5 from Oryza species and P. coarctata showed 4 major amino acid substitutions (140 P/A/T/I, 184 H/R, D332H, V395L), with similarity amongst the tolerant genotypes and the halophyte but in variance with sensitive ones. The best predicted 3D structure of HKT1;5 was generated using Ktrab potassium transporter as template. Among the four substitutions, conserved presence of aspartate (332) and valine (395) in opposite faces of the membrane along the Na+/K+ channel was observed only for the tolerant and halophytic genotypes. A model based on above, as well as molecular dynamics simulation study showed that valine is unable to generate strong hydrophobic network with its surroundings in comparison to leucine due to reduced side chain length. The resultant alteration in pore rigidity increases the likelihood of Na+ transport from xylem sap to parenchyma and further to soil. The model also proposes that the presence of aspartate at the 332 position possibly leads to frequent polar interactions with the extracellular loop polar residues which may shift the loop away from the opening of the constriction at the pore and therefore permit easy efflux of the Na+. These two substitutions of the HKT1;5 transporter probably help tolerant varieties maintain better Na+/K+ ratio for survival under salt stress.

 

See https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6843544/

 

Figure 2: The relative expression levels of HKT1;5. (A) in roots and shoots of Porteresia coarctata, Pokkali, and IR29 under control condition (no additional NaCl). Elongation factor-α (EF-α) used as reference. Values are expressed as mean ± SEM (standard error of mean) and bars indicate SDs. * and ** denoted p 0.05 and p 0.01 respectively as compared to shoot. All expression analysis was performed with six biological and three technical replicates. Next, level of expression of HKT1;5 in different anatomical parts of rice based on (B) mRNAseq data (Supplementary Table 2) and (C) microarray based data (Supplementary Table 3) extracted from Genevestigator expression analysis tools (https://genevestigator.com/gv/) shows preferential expression of HKT1;5 in roots.

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