毛竹PheTCP19b基因可变剪接体对芽发育的影响  

Effect of Alternatively Spliced Variants of Moso Bamboo (Phyllostachys edulis) PheTCP19b Gene on Bud Development

作　　者：郝琴 申孜 闫小玲 周敏舒 郭小勤[1] HAO Qin;SHEN Zi;YAN Xiao-Ling;ZHOU Min-Shu;GUO Xiao-Qin(Bamboo Research Institute/State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学竹子研究院/省部共建亚热带森林培育国家重点实验室,杭州311300

出　　处：《农业生物技术学报》2025年第1期43-54,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32171880);浙江省自然科学基金重点项目(LZ21C160001)。

摘　　要：可变剪接(alternative splicing, AS)是一种转录后的修饰过程,在生物体各种生理过程中发挥重要作用。TCP是一种植物特有的转录因子,参与植物多种生长发育调控过程。为深入探究毛竹(Phyllostachys edulis) TCP19b基因可变剪接体的表达特性及生物学功能,本研究采用RT-PCR技术克隆了毛竹TCP19b基因全长CDS,采用qRT-PCR技术和GUS组织染色分析了可变剪接体的表达量及表达部位,通过亚细胞定位及遗传转化分析了可变剪接体编码蛋白的定位及生物学功能。结果显示,毛竹PheTCP19b基因发生了可变剪接,产生了2种可变剪接体PheTCP19bα和PheTCP19bβ,分别编码346和382个氨基酸,2种可变剪接体都包含保守的bHLH结构域。PheTCP19b与水稻(Oryza sativa) OsTCP19亲缘关系最近。2个可变剪接体的组织特异性表达模式存在一定差异,PheTCP19bα的组织特异性强,而PheTCP19bβ表现出组成型表达特性,油菜素内酯(brassinolide, BR)可诱导2个可变剪接体的表达。2个可变剪接体蛋白均富集在细胞核。PheTCP19bα和PheTCP19bβ均可以将拟南芥(Arabidopsis thaliana) brc1突变体多分枝表型回补到野生型表型。本研究为进一步探讨PheTCP19b基因不同剪接体在芽发育过程中的分子机制提供了基础资料。Alternative splicing(AS)is a post transcriptional modificaption process,and plays important roles in various physiological processes of living organisms.TCP family is a kind of plant specific transcription factor that participates in various growth and development regulatory processes.In order to further explore the expression characteristics and biological function of 2 alternative splice variants of TCP19b gene in moso bamboo(Phyllostachys edulis),RT-PCR was used to clone the full-length CDS of TCP19b from moso bamboo,qRT-PCR and GUS tissue staining were used to analyze the expression level and expression site of the alternative splice variants.The localization and function of alternative splice variants were analyzed by subcellular localization and genetic transformation.The results showed that PheTCP19b gene generated 2 alternative splice variants,PheTCP19bαand PheTCP19bβ,encoding 346 and 382 amino acid,respectively,which contained conserved bHLH domains.The PheTCP19b gene was closely related to rice(Oryza sativa)OsTCP19.There were some differences in tissue-specific expression patterns between the 2 alternative splice variants.The PheTCP19bαhad obvious tissue specificity,while PheTCP19bβshowed constitutive expression pattern.Brassinolide(BR)could induce the expression of 2 alternative splice variants.The 2 alternative splice variants proteins were enriched in the nucleus.Both PheTCP19bαand PheTCP19bβcould complement the multi-branching phenotype of Arabidopsis thaliana brc1 mutant to wild type phenotype.The study provides foundation for further study on the mechanism of PheTCP19b alternative splice variants in bud development.

关 键 词：毛竹 TCP转录因子 可变剪接(AS) 芽发育 油菜素内酯(BR) 

分 类 号：S795.7[农业科学—林木遗传育种]