甘蓝型油菜BnaA02.CPSF6基因的克隆及功能分析  

Cloning and Functional Analysis of the BnaA02.CPSF6 Gene from Brassica napus

作　　者：李青洋 刘翠 何李 彭姗 马嘉吟 胡子祎 刘宏波[1] Qingyang Li;Cui Liu;Li He;Shan Peng;Jiayin Ma;Ziyi Hu;Hongbo Liu(College of Advanced Agricultural Sciences,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学现代农学院,杭州311300

出　　处：《植物学报》2025年第1期62-73,共12页Chinese Bulletin of Botany

基　　金：国家自然科学基金(No.32071929);浙江省自然科学基金(No.LY21C130001);浙江省“十四五”农业新品种选育重大科技专项子课题(No.2021C02064-2-1);浙江农林大学科研发展基金(No.2021FR044)。

摘　　要：CPSF家族蛋白是植物体内mRNA前体中多聚腺苷酸化信号识别、剪切和添加poly(A)的重要因子,对开花时间调控、环境响应和种子发育等具有重要作用。目前,甘蓝型油菜(Brassica napus)CPSF家族基因的功能尚不明确。为探究甘蓝型油菜CPSF家族基因的功能和表达模式,从甘蓝型油菜品种中双11号中克隆得到BnaA02.CPSF6,并对其进行生物信息学、编码蛋白质亚细胞定位、表达模式和基因功能分析。结果表明,BnaA02.CPSF6基因编码区全长1938 bp,编码646个氨基酸残基,无内含子结构,其在甘蓝型油菜中有6个同源基因;BnaA02.CPSF6启动子区存在多个参与光反应的顺式作用元件和MYB结合位点;BnaA02.CPSF6在根、茎、叶、花和不同发育时期种子中均有表达,特别是在发育15–35天的种子中显著高表达,其编码的蛋白定位于细胞核;BnaA02.CPSF6受盐和干旱胁迫诱导上调表达;在ABA、IAA、GA3、SA和MeJA激素处理下,BnaA02.CPSF6基因表达先受到抑制再逐渐恢复至正常水平;在正常条件下,在拟南芥(Arabidopsis thaliana)中过表达BnaA02.CPSF6会出现提前抽薹开花的表型,且莲座叶数量显著减少。综上所述,BnaA02.CPSF6参与非生物胁迫响应并受植物激素调控,可能在开花调控中起促进作用。INTRODUCTION:The CPSF family(cleavage and polyadenylation specificity factor)is a crucial protein family that is responsible for polyadenylation signal recognition in mRNA precursors,cleavage and the addition of poly(A)tails to mRNAs in plants.This family plays crucial roles in the regulation of flowering time,the environmental response,and seed development.Currently,the function of the CPSF family genes in Brassica napus is unclear.RATIONALE:To explore the function and expression patterns of the CPSF gene family,this study cloned BnaA02.CPSF6 from B.napus variety Zhongshuang No.11 and conducted bioinformatics analysis,subcellular localization,expression pattern,and functional characterization of the gene.RESULTS:These results indicate that the coding region of the BnaA02.CPSF6 gene is 1938 bp in length and encodes 646 amino acids without intron structures.Its promoter region contains multiple cis-acting elements involved in light res-ponses and MYB binding sites.Additionally,there are six genes homologous to BnaA02.CPSF6 in B.napus.The BnaA02.CPSF6 gene expressed in the roots,stems,leaves,flowers and different developmental seeds of B.napus,especially significantly higher in 15–35 d developmental seeds,and its encoded protein was localized in the nucleus.The BnaA02.CPSF6 gene expression is upregulated under salt and drought stress.Under treatment with hormones such as ABA,IAA,GA3,SA,and MeJA,the expression of BnaA02.CPSF6 gene is initially inhibited and then gradually recovers to normal levels.Under normal conditions,the overexpression of the BnaA02.CPSF6 gene in Arabidopsis thaliana results in an early bolting phenotype,along with a reduced number of rosette leaves.CONCLUSION:In summary,the above results indicate that the BnaA02.CPSF6 is involved in abiotic stress responses,is regulated by phytohormones,and may also play a promoting role in flowering regulation.

关 键 词：甘蓝型油菜 BnaA02.CPSF6 基因功能 表达模式 生长发育 

分 类 号：Q943.2[生物学—植物学] S565.4[农业科学—作物学]