生物技术通报2025，Vol.41Issue(5)：153-164,12.DOI:10.13560/j.cnki.biotech.bull.1985.2024-1249
辣椒CaFD1基因克隆、表达特征及功能验证
Cloning,Expression Characteristics and Functional Verification of the Pepper CaFD1 Gene
摘要
Abstract
[Objective]The aim is to uncover the molecular mechanism of CaFD1 regulatying pepper's flowering time and its function in the response to abiotic stress,thus providing gene resources for pepper molecular breeding.[Method]The AtFD homologous gene CaFD1 was obtained via RT-PCR from the cDNA of the stem of annual pepper'Zunla-1'.Its physical and chemical properties,protein structure,and phylogenetic connections were examined using bioinformatics.The gene's expression traits under different tissues and stress treatments in pepper were analyzed by RT-qPCR.The gene silencing TRV-CaFD1 vector and subcellular localization 35S:CaFD1 vector were constructed and then introduced into pepper and tobacco respectively through Agrobacterium-mediated transformation.[Result]The entire genetic code of CaFD1,extending across 711 bases,was interpreted to form a protein containing 236 amino acids.The predicted molar mass was approximately 22.34 kD and demonstrated an isoelectric point,determined to be 7.71.It was a hydrophilic protein with an aliphatic amino acid index of 70.85 and high thermal stability.CaFD1 protein possessed two conserved domains,namely the basic amino acid region N-x9-R and the leucine zipper region x6-L-x6-L-x6-L.Its secondary structure was dominated by random coil with a minor portion of α-helix.Promoter prediction analysis revealed the existence of diverse cis-acting elements related to hormones and stress on the CaFD1 promoter.RT-qPCR confirmed that CaFD1 was mainly expressed in the stem,followed by the flower,and hardly expressed in the root.Subcellular localization indicated that the CaFD1 protein was localized in the nucleus.In virus-induced gene silencing plants,i.e.,budding days and flowering days in the plants with silenced CaFD1 gene prolonged,while the phenotypes of overexpressing plants were complementary.[Conclusion]CaFD1 is predominantly expressed in the stem,has the function of promoting plant flowering,and responds to drought,high salt,ABA,and low light stress.关键词
辣椒/CaFD1/开花时间调控/基因表达/功能验证/非生物胁迫Key words
pepper/CaFD1/flowering time regulation/gene expression/functional verification/abiotic stress引用本文复制引用
彭绍智,王登科,张祥,戴雄泽,徐昊,邹学校..辣椒CaFD1基因克隆、表达特征及功能验证[J].生物技术通报,2025,41(5):153-164,12.基金项目
国家自然科学基金项目(32372735) （32372735）
