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通过RNA测序对重要经济油料作物芥菜中高温和干旱胁迫调控基因的全球洞察。

Global insights into high temperature and drought stress regulated genes by RNA-Seq in economically important oilseed crop Brassica juncea.

作者信息

Bhardwaj Ankur R, Joshi Gopal, Kukreja Bharti, Malik Vidhi, Arora Priyanka, Pandey Ritu, Shukla Rohit N, Bankar Kiran G, Katiyar-Agarwal Surekha, Goel Shailendra, Jagannath Arun, Kumar Amar, Agarwal Manu

机构信息

Department of Botany, University of Delhi Main Campus, Delhi, 110007, India.

Department of Plant Molecular Biology, University of Delhi South Campus, Delhi, 110021, India.

出版信息

BMC Plant Biol. 2015 Jan 21;15:9. doi: 10.1186/s12870-014-0405-1.

Abstract

BACKGROUND

Brassica juncea var. Varuna is an economically important oilseed crop of family Brassicaceae which is vulnerable to abiotic stresses at specific stages in its life cycle. Till date no attempts have been made to elucidate genome-wide changes in its transcriptome against high temperature or drought stress. To gain global insights into genes, transcription factors and kinases regulated by these stresses and to explore information on coding transcripts that are associated with traits of agronomic importance, we utilized a combinatorial approach of next generation sequencing and de-novo assembly to discover B. juncea transcriptome associated with high temperature and drought stresses.

RESULTS

We constructed and sequenced three transcriptome libraries namely Brassica control (BC), Brassica high temperature stress (BHS) and Brassica drought stress (BDS). More than 180 million purity filtered reads were generated which were processed through quality parameters and high quality reads were assembled de-novo using SOAPdenovo assembler. A total of 77750 unique transcripts were identified out of which 69,245 (89%) were annotated with high confidence. We established a subset of 19110 transcripts, which were differentially regulated by either high temperature and/or drought stress. Furthermore, 886 and 2834 transcripts that code for transcription factors and kinases, respectively, were also identified. Many of these were responsive to high temperature, drought or both stresses. Maximum number of up-regulated transcription factors in high temperature and drought stress belonged to heat shock factors (HSFs) and dehydration responsive element-binding (DREB) families, respectively. We also identified 239 metabolic pathways, which were perturbed during high temperature and drought treatments. Analysis of gene ontologies associated with differentially regulated genes forecasted their involvement in diverse biological processes.

CONCLUSIONS

Our study provides first comprehensive discovery of B. juncea transcriptome under high temperature and drought stress conditions. Transcriptome resource generated in this study will enhance our understanding on the molecular mechanisms involved in defining the response of B. juncea against two important abiotic stresses. Furthermore this information would benefit designing of efficient crop improvement strategies for tolerance against conditions of high temperature regimes and water scarcity.

摘要

背景

芥菜变种Varuna是十字花科一种具有重要经济价值的油料作物,在其生命周期的特定阶段易受非生物胁迫影响。迄今为止,尚未有人尝试阐明其转录组在高温或干旱胁迫下的全基因组变化。为了全面了解受这些胁迫调控的基因、转录因子和激酶,并探索与重要农艺性状相关的编码转录本信息,我们采用了下一代测序和从头组装的组合方法来发现与高温和干旱胁迫相关的芥菜转录组。

结果

我们构建并测序了三个转录组文库,即芥菜对照(BC)、芥菜高温胁迫(BHS)和芥菜干旱胁迫(BDS)。生成了超过1.8亿条经过纯度过滤的 reads,这些 reads 经过质量参数处理,高质量 reads 使用 SOAPdenovo 组装器进行从头组装。总共鉴定出77750个独特转录本,其中69245个(89%)得到了高可信度注释。我们建立了一个由19110个转录本组成的子集,这些转录本受高温和/或干旱胁迫的差异调控。此外,还分别鉴定出886个和2834个编码转录因子和激酶的转录本。其中许多对高温、干旱或两种胁迫都有响应。高温和干旱胁迫中上调转录因子数量最多的分别属于热休克因子(HSF)和脱水响应元件结合(DREB)家族。我们还鉴定出239条代谢途径,这些途径在高温和干旱处理过程中受到干扰。对与差异调控基因相关的基因本体分析预测它们参与了多种生物学过程。

结论

我们的研究首次全面发现了高温和干旱胁迫条件下的芥菜转录组。本研究生成的转录组资源将增进我们对芥菜应对两种重要非生物胁迫反应所涉及分子机制的理解。此外,这些信息将有助于设计高效的作物改良策略,以提高对高温和缺水条件的耐受性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a24f/4310166/131edf8e06a9/12870_2014_405_Fig1_HTML.jpg

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