Functional Analysis and Interacting Protein Screening of the GbSKIP Gene in Gossypium barbadense

INTRODUCTION: To investigate the biological functions and potential molecular mechanisms of SKIP/SNW proteins in reg the growth and development of Gossypium barbadense, this study cloned GbSKIP and analyzed its expression patterns in different tissues and under different hormone treatments, thhenotypes of GbSKIP-silenced cotton using VIGS technology, and the proteins interacting with GbSKIP. RATIONALE: We utilized bioinformatics, qRT-PCR, virus-induced gene silencing (VIGS), yeast library screening, and bimolecular fluorescence complementatio analyze the molecular characteristics of GbSKIP, its expression patterns in different tissues and under different hormone treatments, the phenotype of GbSKIP-silenced cotton using VIGS, interacting with GbSKIP. RESULTS: qRT-PCR analysis revealed that the expression of GbSKIP in embryonic axis tissuew-as significantly higher than in root, stem, and leaf tissues, and responded to treatments with planthormones such as IAA, ABA, GA, and SA. Silencing of the GbSKIP gene in cotton plants result-ed in a marked inhibitionshoot apical meristem formation, a cessation of true leaf growth, the pr-esence of only one true leaf at the shoot apex, and a significant reduction in leaf area. Four ca-ndidate interactingroteins of GbSKIP were identified through yeast two-hybrid screening, and theinteraction between GbSKIP and the dormancy/auxin-associated protein GbDRM was further verifi-using the bimolecular fluorescence complementation technique,suggesting that GbSKIP may regulatecotton shoot meristem differentiation and growth through its interaction. CONCLUSION: GbSKIP positively regulates the formation of cotton shoot apical meristems; its silencing leads to defects in shoot apical meristem formation and a significant tion in true leaf area. The interaction between GbSKIP and GbDRM suggests a potential molecular mechanism by which GbSKIP regulates cell differentiation and proliferation in shoot me.