Using next-generation sequencing to detect mutations endowing resistance to pesticides: application to acetolactate-synthase (ALS)-based resistance in barnyard grass, a polyploid grass weed

被引：14

作者：

Delye, Christophe ^{[1
]}

Causse, Romain ^{[1
]}

Gautier, Veronique ^{[2
]}

Poncet, Charles ^{[2
]}

Michel, Severine ^{[1
]}

机构：

[1] INRA, Agroecol UMR1347, F-21000 Dijon, France

[2] INRA, Genet Divers & Ecophysiol Cereales UMR1095, Clermont Ferrand, France

来源：

PEST MANAGEMENT SCIENCE | 2015年 / 71卷 / 05期

关键词：

resistance; pesticide; herbicide; diagnosis; next-generation sequencing; Echinochloa; ACETYL-COA CARBOXYLASE; TARGET-SITE RESISTANCE; HERBICIDE RESISTANCE; INHIBITING HERBICIDES; FUNGICIDE RESISTANCE; CROSS-RESISTANCE; ALS INHIBITORS; MOLECULAR TEST; PCR ASSAYS; ALLELE;

D O I：

10.1002/ps.3818

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

BACKGROUNDNext-generation sequencing (NGS) technologies offer tremendous possibilities for accurate detection of mutations endowing pesticide resistance, yet their use for this purpose has not emerged in crop protection. This study aims at promoting NGS use for pesticide resistance diagnosis. It describes a simple procedure accessible to virtually any scientist and implementing freely accessible programs for the analysis of NGS data. RESULTSThree PCR amplicons encompassing seven codons of the acetolactate-synthase gene crucial for herbicide resistance were sequenced using non-quantified pools of crude DNA extracts from 40 plants in each of 28 field populations of barnyard grass, a polyploid weed. A total of 63959 quality NGS sequence runs were obtained using the 454 technology. Three herbicide-resistance-endowing mutations (Pro-197-Ser, Pro-197-Leu and/or Trp-574-Leu) were identified in seven populations. The NGS results were confirmed by individual plant Sanger sequencing. CONCLUSIONThis work demonstrated the feasibility of NGS-based detection of pesticide resistance, and the advantages of NGS compared with other molecular biology techniques for analysing large numbers of individuals. NGS-based resistance diagnosis has the potential to play a substantial role in monitoring resistance, maintaining pesticide efficacy and optimising pesticide applications. (c) 2014 Society of Chemical Industry

引用

页码：675 / 685

页数：11

共 56 条

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