The Importin-β Protein LbSAD2 Enhances Salt Gland Development and Salt Resistance in the Recretohalophyte Limonium bicolor.

Abstract

Limonium bicolor is a typical recretohalophyte with specialised salt glands that secrete excessive Na⁺ out of the plant. The detailed mechanisms of salt gland development and salt resistance are largely unclear. Here, we investigated the function of the importin-β protein LbSAD2 from L. bicolor. Lines with silenced LbSAD2 expression showed significantly lower salt gland density, salt-secretion ability and salt resistance, whereas LbSAD2 overexpression lines had a greater number of salt glands with an abnormal distribution in the abaxial and leaf adaxial surfaces. A previously uncharacterised hydrophobic protein, Lb2G12077, can bind to the LbSAD2 promoter to inhibit the transcription of LbSAD2 verified by a yeast one-hybrid, electrophoretic mobility shift and dual-luciferase reporter assay. Further functional validation revealed that Lb2G12077 demoted salt gland development and salt resistance of L. bicolor. Moreover, a yeast two-hybrid, bimolecular fluorescence complementation and GST pull-down assays indicated that the hypothetical protein Lb2G12567 interacts with LbSAD2, whose silenced lines also showed significantly reduced salt gland density, salt-secretion ability and salt resistance, suggesting Lb2G12567 played a similar positive role in salt resistance. The current LbSAD2 pathway sheds light on salt gland development and salt resistance in L. bicolor, laying the foundation for increasing salt tolerance in crops.