LbHKT1;1 Negatively Regulates Salt Tolerance of <i>Limonium bicolor</i> by Decreasing Salt Secretion Rate of Salt Glands

The HKT-type proteins have been extensively studied and have been shown to play important roles in long-distance Na⁺ transport, maintaining ion homoeostasis and improving salt tolerance in plants. However, there have been no reports on the types, characteristics and functions of HKT-type proteins in Limonium bicolor, a recretohalophyte species with the typical salt gland structure. In this study, five LbHKT genes were identified in L. bicolor, all belonging to subfamily 1 (HKT1). There are many cis-acting elements related to abiotic/biotic stress response on the promoters of the LbHKT genes. LbHKT1;1 was investigated in detail. Subcellular localization results showed that LbHKT1;1 is targeted to the plasma membrane. Functional analysis in yeast showed that LbHKT1;1 has a higher tolerance than AtHKT1;1 under high Na⁺ conditions. Silencing and overexpression of the LbHKT1;1 gene in L. bicolor showed that LbHKT1;1 negatively regulates salt secretion by the salt glands. Further experiments showed that LbbZIP52 can specifically bind to the ABRE element in the LbHKT1;1 promoter and regulate the expression of the LbHKT1;1 gene and is involved in the negative regulation of the salt secretion capacity of L. bicolor. This study demonstrates for the first time that the HKT-type protein is involved in salt secretion by salt glands and provides a new perspective on the function of HKT-type proteins under salt stress conditions.