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The Na + /H +  antiporters (NHXs) are secondary ion transporters to exchange H +  and transfer the Na +  or K +  across membrane, they play crucial roles during plant development and stress responses. To gain insight into the functional divergence of  NHX  genes in poplar, eight  PtNHX  were identified from  Populus trichocarpa  genome. PtNHXs containing 10 transmembrane helices (TMH) and a hydrophilic C-terminal domain, the TMH compose a hollow cylinder to provide the channel for Na +  and H +  transport. The expression patterns and  cis -acting elements showed that all the  PtNHXs  were response to single or multiple stresses including drought, heat, cold, salinity, MV, and ABA. Both the co-expression network and protein-protein interaction network of PtNHXs implying their functional divergence. Interestingly, although  PtNHX7  and  PtNHX8  were generated by whole genome duplication event, they showed significant differences in expression pattern, protein structure, co-expressed genes, and interacted proteins. Only PtNHX7 interact with CBL and CIPK, indicating PtNHX7 is the primary NHX involved in CBL-CIPK pathway during salt stress responses. Natural variation analysis based on 549  P. trichocarpa  individuals indicated the frequency of SNPs in  PtNHX7  was significantly higher than other  PtNHXs . Our findings provide new insights into the functional divergence of  NHX  genes in poplar.

Expression and integrated network analyses revealed functional divergence of NHX-type Na+/H+ exchanger genes in poplar