A method for DNA extraction and molecular identification of Aphids
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...
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| Language: | English |
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Elsevier
2023-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016123001024 |
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| author | M. Suganthi G. Abirami M. Jayanthi K. Ashok Kumar Kalimuthu Karuppanan Senthilkumar Palanisamy |
| author_facet | M. Suganthi G. Abirami M. Jayanthi K. Ashok Kumar Kalimuthu Karuppanan Senthilkumar Palanisamy |
| author_sort | M. Suganthi |
| collection | DOAJ |
| description | 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 Aphis gossypii. 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 Solanum trilobatum provides insights of pest identification and management. |
| first_indexed | 2024-03-13T03:33:43Z |
| format | Article |
| id | doaj.art-becbd9197dd6435b96acf88bd79b02b8 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-03-13T03:33:43Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-becbd9197dd6435b96acf88bd79b02b82023-06-24T05:17:20ZengElsevierMethodsX2215-01612023-01-0110102100A method for DNA extraction and molecular identification of AphidsM. Suganthi0G. Abirami1M. Jayanthi2K. Ashok Kumar3Kalimuthu Karuppanan4Senthilkumar Palanisamy5Department of Biotechnology, Vels Institute of Science, Technology & Advanced Studies (VISTAS), Pallavaram, Tamil Nadu 600117, India; Corresponding authors.Department of Biotechnology, Vels Institute of Science, Technology & Advanced Studies (VISTAS), Pallavaram, Tamil Nadu 600117, IndiaDepartment of Biotechnology, Vels Institute of Science, Technology & Advanced Studies (VISTAS), Pallavaram, Tamil Nadu 600117, IndiaDepartment of Biotechnology, Vels Institute of Science, Technology & Advanced Studies (VISTAS), Pallavaram, Tamil Nadu 600117, IndiaDepartment of Biotechnology, SRM Institute of Science and Technology, Kattankulathur, Chennai, Tamil Nadu 603203, IndiaDepartment of Genetic Engineering, SRM Institute of Science and Technology, Kattankulathur, Chennai, Tamil Nadu 603203, India; Corresponding authors.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 Aphis gossypii. 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 Solanum trilobatum provides insights of pest identification and management.http://www.sciencedirect.com/science/article/pii/S2215016123001024DNA Extraction and PCR amplification of mtCOI gene |
| spellingShingle | M. Suganthi G. Abirami M. Jayanthi K. Ashok Kumar Kalimuthu Karuppanan Senthilkumar Palanisamy A method for DNA extraction and molecular identification of Aphids MethodsX DNA Extraction and PCR amplification of mtCOI gene |
| title | A method for DNA extraction and molecular identification of Aphids |
| title_full | A method for DNA extraction and molecular identification of Aphids |
| title_fullStr | A method for DNA extraction and molecular identification of Aphids |
| title_full_unstemmed | A method for DNA extraction and molecular identification of Aphids |
| title_short | A method for DNA extraction and molecular identification of Aphids |
| title_sort | method for dna extraction and molecular identification of aphids |
| topic | DNA Extraction and PCR amplification of mtCOI gene |
| url | http://www.sciencedirect.com/science/article/pii/S2215016123001024 |
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