水曲柳2个PLT转录因子基因的克隆及表达分析

doi:10.7525/j.issn.1673-5102.2019.01.017

摘要/Abstract

摘要： 为初步探究水曲柳根发育中PLT转录因子的基因功能及遗传调控机理，以水曲柳转录组数据为基础，参考拟南芥AtPLT2和AtPLT3基因序列，通过PCR技术从水曲柳中克隆了两个PLT转录因子基因，通过同源比对将它们命名为FmPLT2和FmPLT3，分别编码532和497个氨基酸残基。生物信息学分析表明，其相对分子质量分别为59和55 kDa，等电点分别为5.98和5.79，均为亲水性不稳定蛋白，均含有2个AP2保守结构域。系统进化分析结果显示，其与同属木犀科的油橄榄OePLT2和OeOLT3转录因子的同源性最高，亲缘关系最近。亚细胞定位预测显示，FmPLT2和FmPLT3蛋白都主要集中于细胞核中。通过qRT-PCR技术分析FmPLT2和FmPLT3在不同组织部位及生根期间的表达情况，结果表明：FmPLT2和FmPLT3具有相似的组织特异性表达，在根中表达含量均最高，在叶中的表达含量均最低；FmPLT2在生根期间的表达量变化极为显著，21天时的表达量是0天时的32倍，但FmPLT3的表达量变化并不显著，表明FmPLT2不仅在根发育中起重要作用，还可能参与细胞增殖和生长等多个发育途径。

关键词: 水曲柳, PLT转录因子, 基因克隆, 表达分析, 根发育

Abstract: In order to investigate the function and mechanism of the PLT transcription factor in the root development of Fraxinus mandshurica, we cloned two PLT transcription factors from F.mandshurica, and named them FmPLT2 and FmPLT3 by homologous alignment, encoding 532 and 497 amino acid residues, respectively. By bioinformatics analysis, the relative molecular masses of FmPLT2 and FmPLT3 were 59 and 55 kDa, respectively, and the isoelectric points were 5.98 and 5.79, respectively. They were all hydrophilic unstable proteins, and both contained two AP2 conserved domains. By phylogenetic analysis, FmPLT2 and FmPLT3 had the highest homology with the OePLT2 and OeOLT3 genes of the Oleaceae, respectively. Subcellular localization predictions showed that both FmPLT2 and FmPLT3 proteins were mainly concentrated in the nucleus. The expression of FmPLT2 and FmPLT3 in different tissues and roots of different developmental stages was analyzed by qRT-PCR. The tissue-specific expression of FmPLT2 and FmPLT3 was similar and they all have the highest expression in roots and the lowest expression in leaves. The expression level of FmPLT2 in roots at different developmental stages is extremely significant. Especially, the expression level of FmPLT2 on the 21st day of root development is 32 times as high as that at the 0 day, but the expression of FmPLT3 was not significant, indicating that FmPLT2 not only plays an important role in root development, but also may participate in multiple developmental pathways such as cell proliferation and growth.

Key words: Fraxinus mandshurica, PLT transcription factors, gene cloning, expression analysis, root development

中图分类号:

S722.3+7

丁一巍, 詹亚光, 张佳薇, 何利明. 水曲柳2个PLT转录因子基因的克隆及表达分析[J]. 植物研究, 2019, 39(1): 139-147.

DING Yi-Wei, ZHAN Ya-Guang, ZHANG Jia-Wei, HE Li-Ming. Cloning and Expression Analysis of Two PLT Transcription Factors Genes in Fraxinus mandshurica[J]. Bulletin of Botanical Research, 2019, 39(1): 139-147.

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