盐碱胁迫下青稞全转录组分析  

Transcriptome Analysis of Tibetan Hulless Barley Under Salt-alkali Stress

作　　者：扎桑 王玉林[1,2] 原红军 杨春葆[1,2] 徐齐君 ZHA Sang;WANG Yu-lin;YUAN Hong-jun;YANG Chun-bao;XU Qi-jun(State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement,Tibet Lhasa 850002,China;Institute of Agricultural Research,Tibet Academy of Agricultural and Animal Husbandry Sciences,Tibet Lhasa 850002,China)

机构地区：[1]省部共建青稞和牦牛种质资源与遗传改良国家重点实验室,西藏拉萨850002 [2]西藏自治区农牧科学院农业研究所,西藏拉萨850002

出　　处：《西南农业学报》2021年第7期1375-1385,共11页Southwest China Journal of Agricultural Sciences

基　　金：西藏财政专项(2017CZZX002);国家大麦青稞产业技术体系(CARS-05)。

摘　　要：【目的】长链非编码RNA(lncRNAs)在动植物的各种生物过程中起着重要作用。青稞是青藏高原的主要粮食作物,对其胁迫诱导基因的研究不仅有助于了解胁迫耐受的分子机制,而且有助于利用基因工程培育胁迫耐受性品种。【方法】本文对2个青稞品种(0119盐碱高抗,0226盐碱敏感)幼苗分别进行盐碱胁迫和对照处理2、8、24、48和72 h后,提取植株叶片RNA进行全转录组测序(设置3个生物学重复),共获得60组RNA-seq数据。【结果】青稞基因组中含有大量的lncRNAs;总共鉴定出6704个lncRNAs,包括2741个基因间长链非编码RNA和1378个反义长链非编码转录本。比较盐碱处理组和对照组的基因表达水平,获得5个时间点的mRNAs和lncRNAs的差异表达基因集,但5个差异表达基因集合之间重叠较少,说明在盐碱胁迫的不同阶段,不同的mRNAs和lncRNAs参与了盐碱响应。研究发现,在24 h时0119和0226中差异表达基因的数目均降低,这可能与作物对刺激反应的两个阶段相关。根据基因的相对位置和表达相关性,预测了lncRNAs的顺式和反式靶基因;功能富集分析发现在0119品种中特异性差异表达的3118个(1303个上调和1815个下调)mRNAs和798个(346个上调和452个下调)lncRNAs可能与耐盐碱性有关,它们直接或间接参与"胁迫应答/刺激反应"、"离子运输"、"膜"和"植物激素信号转导"。为了验证利用RNA-Seq数据计算基因表达量的可靠性,挑选部分mRNA和lncRNA在8,24和48 h盐碱胁迫的0119品种中进行RT-qPCR验证,结果表明RT-qPCR与RNA-seq的基因表达水平一致。【结论】本研究有助于进一步了解青稞盐碱耐受性的机制,提高青稞对盐碱胁迫的耐受性。【Objective】Long non-coding RNAs(lncRNAs)play important roles in various biological processes in animals and plants.Qingke(Tibetan hulless barley)is a staple food in the Tibetan Plateau.The study of stress-inducible genes in Qingke is important to understand the molecular mechanisms of stress tolerance and for breeding stress-tolerant plants via genetic engineering【Method】Sixty ribosomal-RNA-depleted RNA-seq datasets were generated using the leaves of two cultivars of Qingke with different tolerances under 2,8,24,48,72 hours salt-alkali stress treatments and control conditions.【Result】We demonstrated that there was a large number of lncRNAs in the Qingke genome.In total,6704 lncRNAs were identified,including 2741 long intergenic non-coding RNAs and 1378 long non-coding antisense transcripts.Several differentially expressed gene sets of mRNAs and lncRNAs were obtained at five time points under salt-alkali stimulation,and there was little overlap between the five gene sets;thus,different mRNAs and lncRNAs were involved in the response to stress at different stages of salt-alkali stress.The number of differentially expressed genes were decreased at 24 hours in both cultivars,and this time point may separate the two-phase response to stress.Based on the relative position and expression correlation,cis and trans target genes of the lncRNAs were predicted.Especially,3118(1303 and 1815 up-and down-regulated,respectively)mRNAs and 798(346 and 452 up-and down-regulated,respectively)lncRNAs specifically differentially expressed in 0119 cultivar are possibly associated with salt-alkali tolerance based on functional enrichment analysis and are directly or indirectly mapped to‘response to stress/stimulus’,‘ion transport’,‘membrane’and‘plant hormone signal transduction’.The expression pattern of some mRNAs and lncRNAs were validated by quantitative real-time-PCR(qRT-PCR)at 8,24 and 48 hours for salt-alkali stress in 0119 cultivar.【Conclusion】Our work may help to understand the mechanism of salt-a

关 键 词：青稞 盐碱胁迫 长链非编码RNA 转录组 

分 类 号：S512.3[农业科学—作物学]