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差异表达基因在谷子(Setaria italica L.)脱水胁迫过程中的比较转录组分析。

Comparative transcriptome analysis of differentially expressed genes in foxtail millet (Setaria italica L.) during dehydration stress.

机构信息

National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi 110 067, India.

出版信息

Biochem Biophys Res Commun. 2010 Mar 19;393(4):720-7. doi: 10.1016/j.bbrc.2010.02.068. Epub 2010 Feb 18.

Abstract

Dehydration stress is one of the most important abiotic stresses that adversely influence crop growth and productivity. With the aim to understand the molecular mechanisms underlying dehydration stress tolerance in foxtail millet (Setaria italica L.), a drought tolerant crop, we examined its transcriptome changes at two time points (early and late) of dehydration stress. Two suppression subtractive hybridization (SSH) forward libraries were constructed from 21-day old seedlings of tolerant cv. Prasad at 0.5 and 6h PEG-induced dehydration stress. A total of 327 unique ESTs were identified from both libraries and were classified into 11 different categories according to their putative functions. The plant response against dehydration stress was complex, representing major transcripts involved in metabolism, stress, signaling, transcription regulation, translation and proteolysis. By Reverse Northern (RN) technique we identified the differential expression pattern of 327 transcripts, 86 (about 26%) of which showed > or = 1.7-fold induction. Further the obtained results were validated by quantitative real-time PCR (qRT-PCR) to have a comparative expression profiling of randomly chosen 9 up-regulated transcripts (> or =2.5 fold induction) between cv. Prasad (tolerant) and cv. Lepakshi (sensitive) upon dehydration stress. These transcripts showed a differential expression pattern in both cultivars at different time points of stress treatment as analyzed by qRT-PCR. The possible relationship of the identified transcripts with dehydration tolerance mechanism is discussed.

摘要

脱水胁迫是影响作物生长和产量的最重要非生物胁迫之一。为了了解谷子(Setaria italica L.)这种耐旱作物对脱水胁迫耐受的分子机制,我们在 21 天大的幼苗期对其进行了 0.5 和 6 小时 PEG 诱导的脱水胁迫,并在两个时间点(早期和晚期)检测其转录组变化。从耐受品种 Prasad 的幼苗中构建了两个抑制性消减杂交(SSH)正向文库。从两个文库中共鉴定出 327 个独特的 ESTs,并根据其假定功能分为 11 个不同类别。植物对脱水胁迫的反应很复杂,代表了与代谢、应激、信号转导、转录调控、翻译和蛋白水解有关的主要转录物。通过反转Northern(RN)技术,我们鉴定了 327 个转录物的差异表达模式,其中 86 个(约 26%)的表达水平上调了 1.7 倍以上。进一步通过定量实时 PCR(qRT-PCR)验证了随机选择的 9 个上调转录物(>或=2.5 倍诱导)在 cv. Prasad(耐受)和 cv. Lepakshi(敏感)之间的比较表达谱,这些转录物在不同时间点的胁迫处理中在两个品种中表现出不同的表达模式。通过 qRT-PCR 分析,讨论了所鉴定的转录物与脱水耐受机制的可能关系。

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