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拟南芥 TRANSPLANTA 系的集合:基于条件过表达的转录因子功能分析的资源。

The TRANSPLANTA collection of Arabidopsis lines: a resource for functional analysis of transcription factors based on their conditional overexpression.

机构信息

Instituto de Biología Molecular y Celular de Plantas, Valencia (CSIC-UPV), CPI, Edificio 8E, Av. Fausto Elio s/n, 46022, Valencia, Spain.

出版信息

Plant J. 2014 Mar;77(6):944-53. doi: 10.1111/tpj.12443. Epub 2014 Feb 27.

Abstract

Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a β-estradiol-inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS-GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon β-estradiol treatment. As a proof of concept for the exploitation of this resource, β-estradiol-induced proliferation of root hairs, dark-induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full-length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA-mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl-root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution.

摘要

转录因子(TFs)是所有生物中基因表达的关键调节剂。在真核生物中,TFs 通常由功能冗余的大型基因家族成员代表。过表达可能是揭示冗余 TFs 生物学功能的一种手段;然而,TFs 的组成型过表达经常导致严重的发育缺陷,从而阻止其功能表征。条件过表达策略有助于克服这个问题。在这里,我们报告了拟南芥线系 TRANSPLANTA 集合,每个线系都在β-雌二醇诱导启动子的控制下表达 949 个 TFs 之一。到目前为止,已经为 634 个 TF 的诱导表达产生了 1636 个独立的纯合系,平均每个 TF 有 2.6 个系。随着 GUS-GFP 报告基因的使用,随机选择的 TRANSPLANTA 线系在经过β-雌二醇处理后,其条件性转基因表达得到了测试和证实。作为利用该资源的概念验证,在条件表达全长 cDNA 编码 RHD6、WRKY22、MYB123/TT2 和 MYB26 的线系中观察到了根毛的β-雌二醇诱导增殖、暗诱导衰老、花青素积累和矮化,这与这些 TFs 先前报道的表型一致。在用其他 TRANSPLANTA 线系进行的进一步筛选中,鉴定了参与不同植物生物学过程的 TFs,说明了该集合是 TFs 功能表征的有力资源。例如,ANAC058 和一个 TINY/AP2 TF 被鉴定为 ABA 介导的萌发潜能的调节剂,RAP2.10/DEAR4 被鉴定为下胚轴-根过渡区细胞死亡的调节剂。TRANSPLANTA 线系的种子已被存放在诺丁汉拟南芥库存中心,以供进一步分发。

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