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甘蓝型油菜 C2H2 锌指蛋白基因家族的全基因组调查、结构分类和表达分析

Comprehensive genomic survey, structural classification and expression analysis of C2H2 zinc finger protein gene family in Brassica rapa L.

机构信息

Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, College of Crop Science, Fujian Agriculture and Forestry University, Fuzhou, China.

State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Life Sciences, Key Lab of Biopesticides and Chemical Biology, Fujian Agriculture and Forestry University, Fuzhou, China.

出版信息

PLoS One. 2019 May 6;14(5):e0216071. doi: 10.1371/journal.pone.0216071. eCollection 2019.

Abstract

C2H2 zinc finger protein (ZFP) genes have been extensively studied in many organisms and can function as transcription factors and be involved in many biological processes including plant growth and development and stress responses. In the current study, a comprehensive genomics analysis of the C2H2-ZFP genes in B. rapa was performed. A total of 301 B. rapa putative C2H2-ZFP (BrC2H2-ZFP) genes were identified from the available Brassica genome databases, and further characterized through analysis of conserved amino acid residues in C2H2-ZF domains and their organization, subcellular localization, phylogeny, additional domain, chromosomal location, synteny relationship, Ka/Ks ratio, and expression pattern. We also analyzed the expression patterns of eight B. rapa C2H2-ZFP genes under salt and drought stress conditions by using qRT-PCR technique. Our results showed that about one-third of these B. rapa C2H2-ZFP genes were originated from segmental duplication caused by the WGT around 13 to 17 MYA, one-third of them were highly and consecutively expressed in all tested tissues, and 92% of them were located in nucleus by prediction supporting then their functional roles as transcription factors, of which some may play important roles in plant growth and development. The Ka/Ks ratios of 264 orthologous C2H2-ZFP gene pairs between A. thaliana and B. rapa were all, except two, inferior to 1 (varied from 0.0116 to 1.4919, with an average value of 0.3082), implying that these genes had mainly experienced purifying selection during species evolution. The estimated divergence times of the same set of gene pairs ranged from 6.23 to 38.60 MY, with an average value of 18.29 MY, indicating that these gene members have undergone different selective pressures resulting in different evolutionary rates during species evolution. In addition, a few of these B. rapa C2H2-ZFPs were shown to be involved in stress responses in a similar way as their orthologs in A. thaliana. Comparison between A. thaliana and B. rapa orthologous C2H2-ZFP genes showed that the majority of these C2H2-ZFP gene members encodes proteins with conserved subcellular localization and functional domains between the two species but differed in their expression patterns in five tissues or organs. Thus, our study provides valuable information for further functional determination of each C2H2-ZFP gene across the Brassica species, and may help to select the appropriate gene targets for further in-depth studies, and genetic engineering and improvement of Brassica crops.

摘要

C2H2 锌指蛋白 (ZFP) 基因在许多生物体中得到了广泛研究,它们可以作为转录因子发挥作用,并参与包括植物生长发育和应激反应在内的许多生物学过程。在本研究中,对油菜的 C2H2-ZFP 基因进行了全面的基因组分析。从现有的芸薹属基因组数据库中鉴定出了 301 个油菜假定的 C2H2-ZFP(BrC2H2-ZFP)基因,并通过分析 C2H2-ZF 结构域中保守的氨基酸残基及其组织、亚细胞定位、系统发育、附加结构域、染色体定位、基因同线性关系、Ka/Ks 比值和表达模式对其进行了进一步的表征。我们还通过 qRT-PCR 技术分析了 8 个油菜 C2H2-ZFP 基因在盐和干旱胁迫条件下的表达模式。结果表明,这些油菜 C2H2-ZFP 基因中有三分之一左右是由大约 13 到 17 百万年前的 WGT 引起的片段重复产生的,其中三分之一在所有测试的组织中高度连续表达,92%的基因预测定位于细胞核,支持它们作为转录因子的功能作用,其中一些可能在植物生长发育中发挥重要作用。拟南芥和油菜的 264 对同源 C2H2-ZFP 基因对的 Ka/Ks 比值均小于 1(范围为 0.0116 到 1.4919,平均值为 0.3082),表明这些基因在物种进化过程中主要经历了纯化选择。同一组基因对的估计分歧时间范围为 6.23 到 38.60 百万年,平均值为 18.29 百万年,表明这些基因成员在物种进化过程中经历了不同的选择压力,导致不同的进化速率。此外,一些油菜 C2H2-ZFPs 被证明以类似于拟南芥同源物的方式参与应激反应。拟南芥和油菜的同源 C2H2-ZFP 基因的比较表明,这些 C2H2-ZFP 基因成员的大多数编码蛋白在这两个物种中具有保守的亚细胞定位和功能结构域,但在五个组织或器官中的表达模式不同。因此,我们的研究为进一步研究芸薹属物种中每个 C2H2-ZFP 基因的功能提供了有价值的信息,并可能有助于选择适当的基因靶标进行进一步的深入研究和芸薹属作物的遗传工程和改良。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a081/6502316/77b09c087ce9/pone.0216071.g001.jpg

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