Selection of relatively exact reference genes for gene expression studies in goosegrass (Eleusine indica) under herbicide stress.
Selection of relatively exact reference genes for gene expression studies in goosegrass (Eleusine indica) under herbicide stress.
复制标题
除草剂胁迫下牛筋草基因表达研究相对精确内参基因的选择
DOI：
10.1038/srep46494
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发表时间：
2017-04-21
影响因子：
4.6
通讯作者：
Zhang C
中科院分区：
文献类型：
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作者：
Chen J;Huang Z;Huang H;Wei S;Liu Y;Jiang C;Zhang J;Zhang C
鹅观草是世界范围内危害最严重的一年生杂草之一，其进化的抗除草剂种群较难控制。实时定量聚合酶链式反应(QPCR)是研究鹅草抗病机制的常用技术，但目前尚无系统筛选鹅草稳定参考基因的报道。本研究拟对9个候选参考基因在不同组织、不同发育阶段和3种除草剂胁迫下的表达稳定性进行检测。结果表明，对于不同发育阶段和不同器官(对照)，真核细胞启动因子4 A(eIF-4)是最稳定的参考基因。叶绿体乙酰乳酸合成酶(ALS)是草甘膦胁迫下最稳定的参考基因。在草甘膦胁迫下，eIF-4是最好的参考基因。泛素结合酶(UCE)是精喹禾灵胁迫下最稳定的参考基因。EIF-4基因是三种除草剂胁迫下鹅茅的推荐参考基因。此外，配对分析表明，在三种除草剂处理下，7个参考基因足以使基因表达数据标准化。本研究为鹅茅基因转录标准化提供了可靠的参考基因清单，为研究该杂草的抗性机制奠定了基础。
Goosegrass (Eleusine indica) is one of the most serious annual grassy weeds worldwide, and its evolved herbicide-resistant populations are more difficult to control. Quantitative real-time PCR (qPCR) is a common technique for investigating the resistance mechanism; however, there is as yet no report on the systematic selection of stable reference genes for goosegrass. This study proposed to test the expression stability of 9 candidate reference genes in goosegrass in different tissues and developmental stages and under stress from three types of herbicide. The results show that for different developmental stages and organs (control), eukaryotic initiation factor 4 A (eIF-4) is the most stable reference gene. Chloroplast acetolactate synthase (ALS) is the most stable reference gene under glyphosate stress. Under glufosinate stress, eIF-4 is the best reference gene. Ubiquitin-conjugating enzyme (UCE) is the most stable reference gene under quizalofop-p-ethyl stress. The gene eIF-4 is the recommended reference gene for goosegrass under the stress of all three herbicides. Moreover, pairwise analysis showed that seven reference genes were sufficient to normalize the gene expression data under three herbicides treatment. This study provides a list of reliable reference genes for transcript normalization in goosegrass, which will facilitate resistance mechanism studies in this weed species.