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利用生物测定法和 DNA 测序技术对澳大利亚田间烟粉虱种群进行杀虫剂抗性筛选。

Screening for insecticide resistance in Australian field populations of Bemisia tabaci (Hemiptera: Aleyrodidae) using bioassays and DNA sequencing.

机构信息

Faculty of Science and Technology, University of Canberra, Canberra, Australia.

CSIRO, Acton.

出版信息

Pest Manag Sci. 2022 Aug;78(8):3248-3259. doi: 10.1002/ps.6906. Epub 2022 Jun 3.

Abstract

BACKGROUND

Species within the Bemisia tabaci cryptic species complex can cause significant crop damage. We used high-throughput amplicon sequencing to identify the species composition and resistance allele genotypes in field populations from cotton fields in Australia. For selected populations, the resistance phenotype was determined in bioassays and compared with sequencing data.

RESULTS

A metabarcoding approach was used to analyse the species composition in 144 field populations collected between 2013 and 2021. Two mixed AUS I and MEAM1 populations were detected, whereas the remaining 142 populations consisted of MEAM1 only. High-throughput sequencing of organophosphate and pyrethroid resistance gene amplicons showed that the organophosphate resistance allele F331W was fixed (> 99%) in all MEAM1 populations, whereas the pyrethroid resistance allele L925I in the voltage-gated sodium channel gene was detected at varying frequencies [1.0%-7.0% (43 populations); 27.7% and 42.1% (two populations); 95%-97.5% (three populations)]. Neither organophosphate nor pyrethroid resistance alleles were detected in the AUS I populations. Pyrethroid bioassays of 85 MEAM1 field-derived populations detected no resistance in 51 populations, whereas 32 populations showed low frequency resistance, and 2 populations were highly resistant.

CONCLUSIONS

We demonstrate that high-throughput sequencing and bioassays are complementary approaches. The detection of target site mutations and the phenotypic provides a comprehensive analysis of the low-level resistance to pyrethroids that is present in Australian cotton farms. By contrast, a limited survey of whitefly populations from horticulture found evidence of high-level resistance against pyrethroids. Furthermore, we found that the F331W allele (linked to organophosphate resistance) is ubiquitous in Australian MEAM1. © 2022 Commonwealth of Australia. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

摘要

背景

烟粉虱隐种复合种中的物种会对作物造成严重损害。我们使用高通量扩增子测序来鉴定澳大利亚棉田田间种群的物种组成和抗性等位基因基因型。对于选定的种群,在生物测定中测定了抗性表型,并将其与测序数据进行了比较。

结果

使用代谢条码分析方法分析了 2013 年至 2021 年间采集的 144 个田间种群的物种组成。检测到两个混合的 AUS I 和 MEAM1 种群,而其余 142 个种群仅由 MEAM1 组成。有机磷和拟除虫菊酯抗性基因扩增子的高通量测序显示,所有 MEAM1 种群中的有机磷抗性等位基因 F331W 均固定(>99%),而电压门控钠离子通道基因中的拟除虫菊酯抗性等位基因 L925I 的频率则不同[1.0%-7.0%(43 个种群);27.7%和 42.1%(两个种群);95%-97.5%(三个种群)]。AUS I 种群中未检测到有机磷或拟除虫菊酯抗性等位基因。对 85 个 MEAM1 田间衍生种群进行的拟除虫菊酯生物测定中,51 个种群未检测到抗性,32 个种群显示低频率抗性,2 个种群高度抗性。

结论

我们证明了高通量测序和生物测定是互补的方法。靶标位点突变和表型的检测提供了对澳大利亚棉田存在的拟除虫菊酯低度抗性的综合分析。相比之下,对园艺中粉虱种群的有限调查发现了对拟除虫菊酯的高水平抗性的证据。此外,我们发现澳大利亚 MEAM1 中普遍存在与有机磷抗性相关的 F331W 等位基因。©2022 英联邦澳大利亚。害虫管理科学由 John Wiley & Sons Ltd 代表化学工业协会出版。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b567/9546000/805d613ab62b/PS-78-3248-g001.jpg

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