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鉴定毛竹(Phyllostachys edulis)中的 TCP 家族和转拟南芥中 PheTCP9 的耐盐性分析。

Identification of TCP family in moso bamboo (Phyllostachys edulis) and salt tolerance analysis of PheTCP9 in transgenic Arabidopsis.

机构信息

Laboratory of Modern Biotechnology, School of Forestry and Landscape Architecture, Anhui Agricultural University, Hefei, 230036, China.

出版信息

Planta. 2022 Jun 7;256(1):5. doi: 10.1007/s00425-022-03917-z.

Abstract

Bioinformatic analysis of moso bamboo TEOSINTE BRANCHED 1, CYCLOIDEA, and PROLIFERATING CELL FACTORS (TCP) transcription factors reveals their conservation and variation as well as the probable biological functions in abiotic stress response. Overexpressing PheTCP9 in Arabidopsis thaliana illustrates it may exhibit a new vision in different aspects of response to salt stress. Plant specific TCPs play important roles in plant growth, development and stress response, but studies of TCP in moso bamboo are limited. Therefore, in this study, a total of 40 TCP genes (PheTCP1 ~ 40) were identified and characterized from moso bamboo genome and divided into three different subfamilies, namely, 7 in TEOSINTE BRANCHED 1 / CYCLOIDEA (TB1/CYC), 14 in CINCINNATA (CIN) and 19 in PROLIFERATING CELL FACTOR (PCF). Subsequently, we analyzed the gene structures and conserved domain of these genes and found that the members from the same subfamilies exhibited similar exon/intron distribution patterns. Selection pressure and gene duplication analysis results indicated that PheTCP genes underwent strong purification selection during evolution. There were many cis-elements related to phytohermone and stress responsive existing in the upstream promoter regions of PheTCP genes, such as ABRE, CGTCA-motif and ARE. Subcellular localization experiments showed that PheTCP9 was a nuclear localized protein. As shown by β-glucuronidase (GUS) activity, the promoter of PheTCP9 was significantly indicated by salt stress. PheTCP9 was significantly induced in the roots, stems and leaves of moso bamboo. It was also significantly induced by NaCl solution. Overexpressing PheTCP9 increased the salt tolerance of transgenic Arabidopsis. Meanwhile, HO and malondialdehyde (MDA) contents were significantly lower in PheTCP9 over expression (OE) transgenic Arabidopsis than WT. Catalase (CAT) activity, K/Na ratio as well as CAT2 expression level was also much improved in transgenic Arabidopsis than WT under salt conditions. In addition, PheTCP9 OE transgenic Arabidopsis held higher survival rates of seedlings than WT under NaCl conditions. These results showed the positive regulation functions of PheTCP9 in plants under salt conditions.

摘要

毛竹 TEOSINTE BRANCHED 1、CYCLOIDEA 和 PROLIFERATING CELL FACTORS(TCP)转录因子的生物信息学分析揭示了它们在非生物胁迫反应中的保守性和变异性,以及可能的生物学功能。在拟南芥中过表达 PheTCP9 表明它可能在盐胁迫反应的不同方面表现出一种新的现象。植物特异性 TCP 在植物生长、发育和应激反应中发挥重要作用,但毛竹 TCP 的研究有限。因此,在这项研究中,从毛竹基因组中鉴定和表征了总共 40 个 TCP 基因(PheTCP1~40),并将它们分为三个不同的亚家族,即 7 个 TEOSINTE BRANCHED 1/CYCLOIDEA(TB1/CYC)、14 个 CINCINNATA(CIN)和 19 个 PROLIFERATING CELL FACTOR(PCF)。随后,我们分析了这些基因的基因结构和保守结构域,发现来自同一亚家族的成员表现出相似的外显子/内含子分布模式。选择压力和基因复制分析结果表明,PheTCP 基因在进化过程中经历了强烈的纯化选择。在 PheTCP 基因的上游启动子区域存在许多与植物激素和应激反应相关的顺式元件,如 ABRE、CGTCA-motif 和 ARE。亚细胞定位实验表明,PheTCP9 是一种核定位蛋白。β-葡萄糖醛酸酶(GUS)活性显示,PheTCP9 的启动子受盐胁迫显著指示。PheTCP9 在毛竹的根、茎和叶中均有显著诱导。NaCl 溶液也显著诱导 PheTCP9 的表达。过表达 PheTCP9 提高了转基因拟南芥的耐盐性。同时,过表达 PheTCP9 的转基因拟南芥中 HO 和丙二醛(MDA)含量明显低于野生型。在盐胁迫条件下,CAT 活性、K/Na 比值以及 CAT2 表达水平在转基因拟南芥中也得到了显著提高。此外,在 NaCl 条件下,PheTCP9 OE 转基因拟南芥幼苗的存活率高于野生型。这些结果表明 PheTCP9 在植物中对盐胁迫具有正向调控作用。

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