甘蓝型油菜BnaA02CPSF6基因的克隆及功能分析

李青洋, 刘翠, 何李, 彭姗, 马嘉吟, 胡子祎, 刘宏波

doi:10.11983/CBB24068

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DOI: https://doi.org/10.11983/CBB24068

研究报告

甘蓝型油菜BnaA02CPSF6基因的克隆及功能分析

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浙江农林大学现代农学院, 杭州 311300

收稿日期: 2024-05-05

修回日期: 2024-07-19

网络出版日期: 2024-07-29

基金资助

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)和浙江农林大学科研发展基金(No.2021FR044)

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Cloning and Functional Analysis of BnaA02CPSF6 gene from Brassica napus L.

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College of Advanced Agricultural Sciences, Zhejiang A&F University, Hangzhou 311300, China

Received date: 2024-05-05

Revised date: 2024-07-19

Online published: 2024-07-29

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摘要

植物CPSF家族(cleavage and polyadenylation specificity factor)是体内mRNA前体中多聚腺苷酸化信号识别、剪切和添加poly(A)的重要蛋白因子, 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CPSF家族功能尚不明确, 为探究甘蓝型油菜CPSF家族的功能和表达模式, 本研究从甘蓝型油菜品种‘中双11号’中克隆得到BnaA02.CPSF6, 并对其进行生物信息学、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, 无内含子结构, 在甘蓝型油菜中有6个同源基因; BnaA02.CPSF6基因启动子区域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15–35天的发育种子中显著高表达, 其编码的蛋白定位于细胞核; BnaA02.CPSF6基因受盐胁迫和干旱胁迫诱导上调表达; 在ABA、IAA, GA₃, SA, 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, BnaA02.CPSF6基因在拟南芥中过表达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。

关键词： 甘蓝型油菜; BnaA02CPSF6; 基因功能; 表达模式; 生长发育

本文引用格式

李青洋, 刘翠, 何李, 彭姗, 马嘉吟, 胡子祎, 刘宏波 . 甘蓝型油菜BnaA02CPSF6基因的克隆及功能分析[J]. 植物学报, 0 : 0 -0 . DOI: 10.11983/CBB24068

Abstract

The CPSF family (cleavage and polyadenylation specificity factor) is a crucial protein factor that is responsible for polyadenylation signal recognition in mRNA precursors, cleavage and the addition of poly(A) tails to mRNA in plants, it plays a crucial role in the regulation of flowering time, environmental response, and seed development. Currently, the function of the CPSF family in Brassica napus is unclear. To explore the function and expression patterns of the CPSF gene family, this study cloned BnaA02.CPSF6 from B. napus variety 'Zhongshuang 11' and conducted bioinformatics analysis, subcellular localization, expression pattern analysis, and functional characterization of the gene. The results indicate that the coding region of the BnaA02.CPSF6 gene is 1938 bp, encoding 646 amino acids without intron structures. Its promoter region contains multiple cis-acting elements involved in light responses and MYB binding sites. Additionally, there are six homologous genes of BnaA02.CPSF6 in B. napus. The BnaA02.CPSF6 gene is expressed in root, stem, leaf, flower and different developmental seed of B. napus, especially significantly higher in 15-35d developmental seed, and its encoded protein is localized in the nucleus. The BnaA02.CPSF6 gene expression is upregulated under salt and drought stress. Under the treatment of hormones such as ABA, IAA, GA, SA, and MeJA, the expression of the BnaA02.CPSF6 gene is initially inhibited and then gradually recovers to normal levels. Under normal condition, overexpression of the BnaA02.CPSF6 gene in Arabidopsis thaliana results in an early bolting and flowering phenotype, along with a reduced number of rosette leaves. In summary, the above results indicate that the BnaA02.CPSF6 is involved in abiotic stress responses and regulated by phytohormones, may also play a promoting role in flowering regulation.

Key words： Brassica napus; BnaA02CPSF6; Gene function; Expression patterns; Growth and development

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