Xyloglucan endotransglucosylase-hydrolase 22 positively regulates response to cold stress in upland cotton (Gossypium hirsutum L.)

Xyloglucan endotransglucosylase/hydrolases (XTHs) are cell wall modifying enzymes that function in response to multiple abiotic stresses by promoting cell wall reinforcement in plants. However, it is elusive whether GhXTH22 plays a role in cold stress tolerance in upland cotton (Gossypium hirsutum L.). In this study, we found that Arabidopsis (Arabidopsis thaliana (L.) Heynh) transgenic lines overexpressing GhXTH22 exhibit clearly increased cold tolerance compared with wild type (WT). Moreover, cold stress results in markable enhancements in the activities of catalase (CAT) and peroxidase (POD), cell wall rigidity, and the expression of cold responsive genes, as well as a notable reduction in malondialdehyde (MDA) levels in the overexpression (OE) lines compared with those in WT plants. Consistently, silencing GhXTH22 by virus-induced gene silencing (VIGS) method causes markedly decreases in cold stress tolerance in upland cotton. Additionally, by electrophoretic mobility shift assay (EMSA) and dual-luciferase assay, it was confirmed that GhMYB44 activate the expression of GhXTH22 by binding to AACNG motif on the promoter, and silencing the expression of GhMYB44 can decrease the cold resistance of cotton. In summary, this study systematically confirmed the cold resistance function of GhXTH22 and discovered the upstream regulating gene of GhXTH22, providing gene resources for cold resistance and enriching the knowledge on the mechanism of cold resistance.