Suganthi M, Abirami G, Jayanthi M, Kumar K Ashok, Karuppanan Kalimuthu, Palanisamy Senthilkumar
Department of Biotechnology, Vels Institute of Science, Technology & Advanced Studies (VISTAS), Pallavaram, Tamil Nadu 600117, India.
Department of Biotechnology, SRM Institute of Science and Technology, Kattankulathur, Chennai, Tamil Nadu 603203, India.
MethodsX. 2023 Feb 27;10:102100. doi: 10.1016/j.mex.2023.102100. eCollection 2023.
Aphid species (Insecta, Hemiptera) are economically important invasive pest throughout the world, though their identification is intricate due to tiny size and inconspicuous nature of morphology. Mitochondrial cytochrome c oxidase I (mtCOI) region has been proven to be a standard barcode to identify the diverse array of insect groups. Isolation of good quality DNA is a fundamental first step in insect DNA barcoding which is obtained by standardizing the DNA isolation method. In this study, we demonstrate a modified CTAB method for the isolation of DNA to maximize the quality and yield from small aphids. This method will help the researchers to efficiently isolate DNA from small aphid and the method can be utilized for other small insects as well. We evaluated the quality of the isolated DNA and the mtCOI gene region were subjected to PCR amplification. Further, the gene segment was sequenced and gene annotation was done by NCBI BLAST program through which the insect was found to be . This study provides a set of molecular tools that can be used for identification of insect at species level through DNA barcoding and biodiversity analysis.•Detailed method to maximize quality and quantity of genomic DNA isolated from aphids.•Molecular identification of aphids using mtCOI gene amplification and sequence validation.•First report on Aphis gossypii infecting provides insights of pest identification and management.
蚜虫种类（昆虫纲，半翅目）是全球具有重要经济意义的入侵害虫，不过由于其体型微小且形态特征不明显，对它们的鉴定很复杂。线粒体细胞色素c氧化酶I（mtCOI）区域已被证明是鉴定各类昆虫群体的标准条形码。高质量DNA的分离是昆虫DNA条形码技术的基本第一步，这可通过标准化DNA分离方法来实现。在本研究中，我们展示了一种改良的CTAB法用于DNA分离，以从小型蚜虫中最大限度地提高DNA质量和产量。该方法将帮助研究人员从小型蚜虫中高效分离DNA，并且该方法也可用于其他小型昆虫。我们评估了分离出的DNA的质量，并对mtCOI基因区域进行了PCR扩增。此外，对基因片段进行了测序，并通过NCBI BLAST程序进行了基因注释，据此发现该昆虫为 。本研究提供了一套分子工具，可用于通过DNA条形码技术在物种水平上鉴定昆虫以及进行生物多样性分析。•从小型蚜虫中分离出基因组DNA并使其质量和数量最大化的详细方法。•利用mtCOI基因扩增和序列验证对蚜虫进行分子鉴定。•关于棉蚜感染 的首次报道为害虫鉴定和管理提供了见解。
