北京林业大学学报2025，Vol.47Issue(12)：87-96,10.DOI:10.12171/j.1000-1522.20250125
无患子SmCYCD3;2基因的克隆与功能研究
Cloning and functional studies on SmCYCD3;2 gene in Sapindus mukorossi
摘要
Abstract
[Objective]To investigate the role of Sapindus mukorossi SmCYCD3;2 gene in flower sex differentiation,to provide valuable reference genes for improving the female-to-male ratio of S.mukorossi flowers,and then to lay a solid theoretical foundation for improving S.mukorossi fruit yields through molecular design breeding.[Method]The SmCYCD3;2 gene of S.mukorossi was cloned,subjected to multiple sequence comparison and phylogenetic evolutionary analysis,and its expression pattern in different organs was analyzed by real-time fluorescence quantitative PCR(RT-qPCR).We constructed overexpression vectors and heterologously transformed Arabidopsis thaliana to compare its growth phenotypes,stained its mature anthers with Alexandrium staining,observed pollen viability and in vitro pollen germination using the pollen germination assay,and observed the morphology and surface features of anthers and pollen with the aid of scanning electron microscopy.[Result](1)The full length of SmCYCD3;2 gene sequence was 1 092 bp,encoding 364 amino acids,and it is closely related to Populus trichocarpa.(2)RT-qPCR analysis revealed that the expression of SmCYCD3;2 gene was highest in female flowers of S.mukorossi,followed by leaves,and the lower expression was found in male flowers.(3)Compared with wild-type plants,transgenic A.thaliana exhibited a series of significant growth phenotypic changes:growth rate was slowed down,plant height was significantly reduced,and leaves appeared curled and slightly bent at each stem node.In addition,the floral organs phenotype also underwent significant changes,which were characterized by elongated pistils,reduced and shorter stamens,reduced pollen activity,and ultimately shorter fruit pods.(4)In transgenic A.thaliana plants,the expression of cell cycle-related genes(AtCYCDs)and genes promoting gynoecium elongation and development(AtCRC)were significantly up-regulated,while the expression of genes related to stamen development(AtAP3,AtPI,AtAMS,and AtDEL)as well as genes related to stem and leaf development(AtPIN)were significantly down-regulated.[Conclusion]The SmCYCD3;2 gene is able to regulate the development of stem and leaf,promote pistil elongation and inhibit the stamen development,which plays an important role in sex differentiation.The results of the study provide new insights into the functional resolution of CYCD3 gene,and lay a pre-theoretical foundation for improving the female-to-male ratio of flowers and increasing fruit yield of S.mukorossi.关键词
无患子/细胞周期蛋白/性别分化/基因克隆/功能研究/基因表达Key words
Sapindus mukorossi/cell cycle proteins/sex differentiation/gene cloning/functional studies/gene expression分类
农业科技引用本文复制引用
高亚娇,赵国春,贾黎明,陈仲..无患子SmCYCD3;2基因的克隆与功能研究[J].北京林业大学学报,2025,47(12):87-96,10.基金项目
福建省科技计划(2023N3005),国家自然科学基金项目(32371857),国家重点研发计划(2023YFD2201103). （2023N3005）
