Abstract: 【Objective】To identify and analyze the cysteine-rich receptor-like kinase（CRK） gene family members of Gossypium hirsutum,and provide genetic resources and theoretical basis for cotton Verticillium wilt resistance breeding.【Methods】According to the results of whole genome sequencing,the GhCRK gene family in the Gossypium hirsutum Nongdamian 8 genome was identified using bioinformatics methodologies,analyses were conducted on protein physicochemical properties,chromosomal localization and genetic evolution. Furthermore,gene expression data were employed to potential Verticillium wilt resistance genes,and the resistance function of candidate gene was assessed through virus induced gene silencing（VIGS） technology.【Results】The genome of Gossypium hirsutum cultivar Nongdamian 8 harbored a total of fifty GhCRKs,distributed across fourteen chromosomes. These length of the encoded protein was 390 to 1 774 amino acid residues,resulting in diverse molecular weights spanning from 43.15 kDa to 196.66 kDa,the average isoelectric point was observed at 6.94. The majority of GhCRKs were predominantly localized in the plasma membrane. Protein domain analysis showed that 90 % of GhCRKs（45） contained two Stress-antifung domains. The promoter regions of GhCRKs encompassed seven cis-acting elements relating to responsiveness to plant hormones,including methyl jasmonate,abscisic acid,salicylic acid,et al. Fourteen GhCRKs exhibited up regulation expression in response to Verticillium dahliae infection. Following the silencing of GhCRK08 and GhCRK33,Gossypium hirsutum exhibited exacerbated symptoms of Verticillium wilt and a significantly higher disease index.【Conclusion】The genome of Gossypium hirsutum Nongdamian 8 contains a substantial number of CRKs,among which 14 hold potential as significant candidate genes for Verticillium wilt resistance. Additionally,the role of GhCRK08 and GhCRK33 in Verticillium wilt resistance has been further elucidated.

Key words: Gossypium hirsutum, Cysteine-rich receptor-like kinase（CRK） gene family, Verticillium wilt, Bioinformatics, Gene function, Gene expression