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从耐硼植物满天星叶片中鉴定差异表达转录本

Identification of differentially expressed transcripts from leaves of the boron tolerant plant Gypsophila perfoliata L.

作者信息

Unver Turgay, Bozkurt Osman, Akkaya Mahinur S

机构信息

Department of Chemistry, Middle East Technical University, 06531, Ankara, Turkey.

出版信息

Plant Cell Rep. 2008 Aug;27(8):1411-22. doi: 10.1007/s00299-008-0560-7. Epub 2008 May 27.

Abstract

Very recently some of the species of Gypsophila genus collected from the boron rich soils in Turkey were shown to be remarkably tolerant to high levels of boron. A limited amount of boron is necessary for the normal development of plants; however, a high level of boron in soil is generally toxic. Nevertheless, the adaptability of plant species allows them to withstand the presence of extreme amounts of metal ion by various strategies. This study is conducted on highly boron tolerant Gypsophila perfoliata L. collected from a location in the boron mining area. The plant samples were transferred into plant nutritional medium in the presence high; approximately 500 (35 mg/kg), 1,000, and 30 microM (considered normal) boron concentrations. We compared the transcriptome of the plant sample treated with the excess levels of boron to that of the samples grown under normal concentration using differential display PCR (DDRT-PCR) method. Thirty bands showing differential expression levels (presence or absence of bands or varying intensities) in either of approximately 500 or 30 microM B concentrations at varying time points were excised, cloned, and sequenced. Among which, 18 of them were confirmed via quantitative reverse transcription real time PCR (qRT-PCR). We are reporting the first preliminary molecular level study of boron tolerance on this organism by attempting to identify putative genes related in the tolerance mechanism. The gene fragments are consistent with the literature data obtained from a proteomics study and a metabolomics study performed in barley under varying boron concentrations.

摘要

最近,从土耳其富硼土壤中采集的一些丝石竹属植物被证明对高硼水平具有显著的耐受性。植物的正常发育需要有限量的硼;然而,土壤中高含量的硼通常是有毒的。尽管如此,植物物种的适应性使它们能够通过各种策略抵御极端数量金属离子的存在。本研究以从硼矿区某一地点采集的高度耐硼的贯叶丝石竹为对象。将植物样本转移到硼浓度较高(约500(35毫克/千克)、1000和30微摩尔(视为正常))的植物营养培养基中。我们使用差异显示PCR(DDRT-PCR)方法,将用过量硼处理的植物样本的转录组与在正常浓度下生长的样本的转录组进行了比较。在不同时间点,在约500或30微摩尔硼浓度下,有30条显示差异表达水平(条带的存在或缺失或强度变化)的条带被切下、克隆并测序。其中,18条通过定量逆转录实时PCR(qRT-PCR)得到证实。我们通过尝试鉴定与耐受机制相关的推定基因,首次报道了对该生物体硼耐受性的初步分子水平研究。这些基因片段与在不同硼浓度下大麦进行的蛋白质组学研究和代谢组学研究获得的文献数据一致。

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