Reference genes to study herbicide stress response in Lolium sp.: up-regulation of P450 genes in plants resistant to acetolactate-synthase inhibitors.
Reference genes to study herbicide stress response in Lolium sp.: up-regulation of P450 genes in plants resistant to acetolactate-synthase inhibitors.
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DOI：
10.1371/journal.pone.0063576
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Délye C
中科院分区：
文献类型：
--
作者：
Duhoux A;Délye C
许多基因表达的变异是植物对环境胁迫反应的基础。除草剂非靶点抗性（NTSR）是禾本科杂草化学防除的主要威胁，它是由参与除草剂胁迫反应的一组基因控制的。因此，定量PCR分析允许可靠的比较基因表达的关键，以确定基因管理NTSR。这项工作的目的是确定一组稳定表达的参考基因，以用作定量PCR数据的标准化的研究调查NTSR除草剂抑制乙酰乳酸合酶（ALS）的主要禾本科杂草黑麦草属物种的内部标准。基因表达的稳定性进行了评估，在植物抗或敏感的两个ALS抑制剂，进行或不进行除草剂胁迫。使用在程序BestKeeper、Normalization和geNorm中实施的三种互补方法，帽结合蛋白、甘油醛-3-磷酸脱氢酶和泛素被鉴定为最合适的参考基因。该参考基因组可用于研究其他杂草对除草剂的反应。比较了编码两种除草剂靶标酶（ALS和乙酰辅酶A羧化酶）和五种具有潜在除草剂降解活性的细胞色素P450（CYP 14）的基因在ALS抑制剂抗性和敏感植物之间的表达。总的来说，除草剂的应用增强了hepatocyte基因的表达。与敏感植物相比，在抗性植物中观察到的所有p53基因的组成性上调表明抗性植物中的次生代谢增强。全面的转录组研究相关的基因表达分析，使用参考基因集验证这里需要解开NTSR遗传决定因素。
Variation in the expression of numerous genes is at the basis of plant response to environmental stresses. Non-target-site-based resistance to herbicides (NTSR), the major threat to grass weed chemical control, is governed by a subset of the genes involved in herbicide stress response. Quantitative PCR assays allowing reliable comparison of gene expression are thus key to identify genes governing NTSR. This work aimed at identifying a set of reference genes with a stable expression to be used as an internal standard for the normalisation of quantitative PCR data in studies investigating NTSR to herbicides inhibiting acetolactate synthase (ALS) in the major grass weed Lolium sp. Gene expression stability was assessed in plants resistant or sensitive to two ALS inhibitors, subjected or not to herbicide stress. Using three complementary approaches implemented in the programs BestKeeper, NormFinder and geNorm, cap-binding protein, glyceraldehyde-3-phosphate-dehydrogenase and ubiquitin were identified as the most suitable reference genes. This reference gene set can probably be used to study herbicide response in other weed species. It was used to compare the expression of the genes encoding two herbicide target enzymes (ALS and acetyl-coenzyme A carboxylase) and five cytochromes P450 (CYP) with potential herbicide-degrading activity between plants resistant or sensitive to ALS inhibitors. Overall, herbicide application enhanced CYP gene expression. Constitutive up-regulation of all CYP genes observed in resistant plants compared to sensitive plants suggested enhanced secondary metabolism in the resistant plants. Comprehensive transcriptome studies associated to gene expression analyses using the reference gene set validated here are required to unravel NTSR genetic determinants.
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影响因子：
5.3
作者：
Jarosová J;Kundu JK
通讯作者：
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5.2
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