Identification of the High-Affinity Potassium Transporter Gene Family in Perennial Ryegrass (Lolium perenne) and Its Potential Role in Salt Stress.

Background: Perennial ryegrass (Lolium perenne L.), a widely cultivated turfgrass and forage species in Europe and North America, exhibits rapid growth and notable salt tolerance. The high-affinity potassium transporter (HKT) gene family has been implicated in salt stress responses across multiple plant species. However, whether the salt tolerance of L. perenne is closely associated with its HKT gene family remains unclear. Methods and Results: In this study, we systematically identified HKT family members in the L. perenne genome. Five HKT genes were identified and classified into three subfamilies. Among these, LpHKT1a-c exhibited canonical class I features with a conserved serine (S) residue in the P1 domain, whereas LpHKT2 belonged to class II, characterized by a glycine (G) residue in the same domain. Notably, LpHKT3 formed a distinct subfamily with a truncated structure and divergent P1/P2 domains, suggesting potential non-canonical functions. LpHKT1a likely lacked the P4 domain. Promoter analysis revealed that all five LpHKT genes contain multiple stress-related cis-acting elements. Real-time quantitative reverse transcription polymerase chain reaction results showed that LpHKT1b/c and LpHKT2 were highly expressed in both roots and leaves. Under low-concentration NaCl stress (25 mM), the expression of these three genes significantly increased by 8- to 12-fold at 6-12 h post-treatment (vs. control). Ion accumulation analysis demonstrated a rapid increase in Na+ levels following NaCl treatment, whereas K+ concentrations initially remained stable but significantly increased after 24 h. Conclusions: Combined with the cellular localization of LpHKT1c predominantly in the xylem, these findings suggest that LpHKT genes may be involved in Na+ and K+ transport in roots. This study represents the first genome-wide identification of the HKT gene family in L. perenne, providing critical insights into the molecular mechanisms underlying its salt tolerance and offering valuable genetic resources for molecular breeding aimed at enhancing stress resilience.