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通过物种特异性PCR对褐纹蝽寄生蜂进行分子鉴定

Molecular Identification of , Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR.

作者信息

Chen Maple N, Santander Ricardo D, Talamas Elijah J, Jentsch Peter J, Bon Marie-Claude, Aćimović Srđan G

机构信息

Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA.

Hudson Valley Research Laboratory, School of Integrative Plant Sciences, Plant Pathology and Plant-Microbe Biology Section, Cornell University, Highland, NY 12528, USA.

出版信息

Insects. 2021 May 18;12(5):467. doi: 10.3390/insects12050467.

Abstract

The samurai wasp, (Ashmead), has been proposed as a biocontrol agent against brown marmorated stink bugs (BMSB), due to its ability to parasitize and kill BMSB eggs. However, the wasps' small size makes it challenging for those untrained in morphological identification to determine the wasps' species. To circumvent this problem, a molecular method was created to identify . The method uses species-specific primers, designed in this study, which target the variable region of the mitochondrial Cytochrome Oxidase 1 () locus. After confirming successful DNA extraction from samples, the PCR amplification using our primers produced 227-bp PCR products for all specimens and no amplification in other microhymenoptera candidates. Additionally, DNA from BMSB-parasitized eggs gave positive PCR amplification, while the control BMSB samples showed no amplification. This indicates that PCR with our primers specifically and sensitively differentiates specimens from other similar wasp species and discriminates between -parasitized and non-parasitized BMSB eggs. Finally, an in silico analysis of sequences demonstrated that our primers match the sequences of four different haplotypes of , indicating that our diagnostic method could potentially be applied to analyze populations throughout North America, Europe, and parts of Asia.

摘要

武士黄蜂(Ashmead)因其能够寄生并杀死褐边绿刺蛾(BMSB)的卵,已被提议作为对抗褐边绿刺蛾的生物防治剂。然而,黄蜂体型小,对于未经形态学鉴定训练的人来说,确定黄蜂的种类具有挑战性。为了解决这个问题,创建了一种分子方法来鉴定……。该方法使用了本研究中设计的物种特异性引物,这些引物靶向线粒体细胞色素氧化酶1(……)基因座的可变区域。在确认从样本中成功提取DNA后,使用我们的引物进行PCR扩增,所有……标本均产生了227 bp的PCR产物,而其他膜翅目候选物种未出现扩增。此外,来自被BMSB寄生的卵的DNA进行PCR扩增呈阳性,而对照BMSB样本未出现扩增。这表明使用我们的引物进行PCR能够特异性且灵敏地将……标本与其他类似黄蜂物种区分开来,并区分被……寄生和未被寄生的BMSB卵。最后,对……序列的电子分析表明,我们的引物与……四种不同单倍型的序列匹配,这表明我们的诊断方法有可能应用于分析北美、欧洲和亚洲部分地区的……种群。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54aa/8157830/cdac8ea19fbb/insects-12-00467-g001.jpg

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