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鉴定和分析玉米(Zea mays L.)中 FtsH 基因家族的两个成员。

Identification and characterization of two members of the FtsH gene family in maize (Zea mays L.).

机构信息

School of Life Sciences, Shandong University, 27 Shanda South Road, 250100, Jinan, Shandong, China.

出版信息

Mol Biol Rep. 2010 Feb;37(2):855-63. doi: 10.1007/s11033-009-9691-3. Epub 2009 Aug 8.

Abstract

Two full-length cDNAs, designated as ZmFtsH2A and ZmFtsH2B, were isolated from maize (Zea mays L.) by suppression subtractive hybridization coupled with in silico cloning approach. The predicted proteins of ZmFtsH2A and ZmFtsH2B both consisted of 677 amino acid residues and displayed high similarity to FtsH2 protease of Arabidopsis thaliana. DNA gel blotting analysis indicated that AtFtsH2-like genes exist as two copies in maize genome. The genomic sequences of ZmFtsH2A and ZmFtsH2B were cloned and the main difference was that the first intron of ZmFtsH2B was much longer than that of ZmFtsH2A. RT-PCR analysis revealed that both genes were constitutively expressed in all examined tissues and the expression level of ZmFtsH2B transcripts was higher than that of ZmFtsH2A. The responses of the two genes in maize seedlings to PEG, cold, high salt, and ABA treatments were compared, and the results showed that ZmFtsH2B transcription in leaves was markedly up-regulated by water deficit stress and ABA treatments while ZmFtsH2A constitutively expressed both in leaves and roots under all tested stressful conditions. Drought tolerance of transgenic tobaccos overexpressing ZmFtsH2A and ZmFtsH2B weren't improved compared to wild-type controls, which indicated that two genes might not be directly involved in plant drought tolerance or the number of functional FtsH heterocomplex might not be increased in this condition. Our current study provides fundamental information for the further investigation of the maize FtsH proteins.

摘要

通过抑制消减杂交结合计算机克隆方法,从玉米(Zea mays L.)中分离得到两个全长 cDNA,分别命名为 ZmFtsH2A 和 ZmFtsH2B。ZmFtsH2A 和 ZmFtsH2B 的预测蛋白均由 677 个氨基酸残基组成,与拟南芥 FtsH2 蛋白酶具有高度相似性。DNA 凝胶印迹分析表明,AtFtsH2 样基因在玉米基因组中存在两个拷贝。克隆了 ZmFtsH2A 和 ZmFtsH2B 的基因组序列,主要区别在于 ZmFtsH2B 的第一个内含子比 ZmFtsH2A 长得多。RT-PCR 分析表明,两个基因在所有检测的组织中均持续表达,ZmFtsH2B 转录本的表达水平高于 ZmFtsH2A。比较了这两个基因在玉米幼苗对 PEG、冷、高盐和 ABA 处理的反应,结果表明,ZmFtsH2B 在叶片中的转录在水分亏缺胁迫和 ABA 处理下明显上调,而 ZmFtsH2A 在所有测试的胁迫条件下均在叶片和根部持续表达。与野生型对照相比,过表达 ZmFtsH2A 和 ZmFtsH2B 的转基因烟草的耐旱性没有提高,这表明这两个基因可能不直接参与植物耐旱性,或者在这种情况下,功能性 FtsH 异源复合物的数量可能没有增加。我们目前的研究为进一步研究玉米 FtsH 蛋白提供了基础信息。

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