Identification of the cysteine-rich transmembrane module CYSTM family in upland cotton and functional analysis of GhCYSTM5_A in cold and drought stresses.

Abiotic stress poses adverse impacts on cotton production, raising demands for a better understanding of stress-response mechanisms and developing strategies to improve plant performance to cope with stress. CYSTM (Cysteine-rich transmembrane module) is a widely distributed and conserved family in eukaryotes that performs potential functions in stress tolerance. However, CYSTM genes and their role in stress response is uncharacterized in cotton. Herein, we identified a total of 23 CYSTM genes from upland cotton. They underwent mainly segmental duplications and experienced purifying selection during evolution. Expression profiles revealed GhCYSTMs were closely related to abiotic stress response. Furthermore, GhCYSTM5_A overexpression enhanced the cold and drought tolerance of cotton, while RNAi-mediated knockdown of GhCYSTM5_A decreased stress tolerance. Transcriptome analysis revealed GhCYSTM5_A may contribute to cold and drought tolerance by regulating the expression of oxidative stress-related genes through MAPK signaling. GhCYSTM5_A, localized in the nucleus and cytoplasm interacted with a secreted cysteine-rich peptide GhGASA14. Moreover, GhGASA14 silencing rendered cotton plants vulnerable to cold and drought. These results suggested the potential functions of GhCYSTM genes in abiotic stress and a positive role of GhCYSTM5_A in cold and drought tolerance. This study sheds light on comprehensive characteristics of GhCYSTM, and provides candidate genes for genetic breeding.