Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR
The samurai wasp, <i>Trissolcus japonicus</i> (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 ident...
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MDPI AG
2021-05-01
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| Series: | Insects |
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| Online Access: | https://www.mdpi.com/2075-4450/12/5/467 |
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| author | Maple N. Chen Ricardo D. Santander Elijah J. Talamas Peter J. Jentsch Marie-Claude Bon Srđan G. Aćimović |
| author_facet | Maple N. Chen Ricardo D. Santander Elijah J. Talamas Peter J. Jentsch Marie-Claude Bon Srđan G. Aćimović |
| author_sort | Maple N. Chen |
| collection | DOAJ |
| description | The samurai wasp, <i>Trissolcus japonicus</i> (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 <i>T. japonicus</i>. The method uses species-specific primers, designed in this study, which target the variable region of the mitochondrial Cytochrome Oxidase 1 (<i>CO1</i>) locus. After confirming successful DNA extraction from samples, the PCR amplification using our primers produced 227-bp PCR products for all <i>T. japonicus</i> 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 <i>T. japonicus</i> specimens from other similar wasp species and discriminates between <i>T. japonicus</i>-parasitized and non-parasitized BMSB eggs. Finally, an in silico analysis of <i>CO1</i> sequences demonstrated that our primers match the sequences of four different haplotypes of <i>T. japonicus</i>, indicating that our diagnostic method could potentially be applied to analyze <i>T. japonicus</i> populations throughout North America, Europe, and parts of Asia. |
| first_indexed | 2024-03-10T11:17:47Z |
| format | Article |
| id | doaj.art-19904f2ababb4e349c20c0f269a4b985 |
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| issn | 2075-4450 |
| language | English |
| last_indexed | 2024-03-10T11:17:47Z |
| publishDate | 2021-05-01 |
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| record_format | Article |
| series | Insects |
| spelling | doaj.art-19904f2ababb4e349c20c0f269a4b9852023-11-21T20:13:49ZengMDPI AGInsects2075-44502021-05-0112546710.3390/insects12050467Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCRMaple N. Chen0Ricardo D. Santander1Elijah J. Talamas2Peter J. Jentsch3Marie-Claude Bon4Srđan G. Aćimović5Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USAHudson Valley Research Laboratory, School of Integrative Plant Sciences, Plant Pathology and Plant-Microbe Biology Section, Cornell University, Highland, NY 12528, USADepartment of Plant Industry, Florida Department of Agriculture and Consumer Services, Gainesville, FL 32399, USAHudson Valley Research Laboratory, Department of Entomology, Cornell University, Highland, NY 12528, USAEuropean Biological Control Laboratory, United States Department of Agriculture-ARS, 34980 Montferrier-sur-Lez, FranceAlson H. Smith Jr. Agricultural Research and Extension Center, Department of Plant Pathology, Physiology, and Weed Science, Virginia Polytechnic Institute and State University, Winchester, VA 22602, USAThe samurai wasp, <i>Trissolcus japonicus</i> (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 <i>T. japonicus</i>. The method uses species-specific primers, designed in this study, which target the variable region of the mitochondrial Cytochrome Oxidase 1 (<i>CO1</i>) locus. After confirming successful DNA extraction from samples, the PCR amplification using our primers produced 227-bp PCR products for all <i>T. japonicus</i> 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 <i>T. japonicus</i> specimens from other similar wasp species and discriminates between <i>T. japonicus</i>-parasitized and non-parasitized BMSB eggs. Finally, an in silico analysis of <i>CO1</i> sequences demonstrated that our primers match the sequences of four different haplotypes of <i>T. japonicus</i>, indicating that our diagnostic method could potentially be applied to analyze <i>T. japonicus</i> populations throughout North America, Europe, and parts of Asia.https://www.mdpi.com/2075-4450/12/5/467samurai waspPCR identificationspecies-specific primers<i>Halyomorpha halys</i>nondestructive DNA extraction |
| spellingShingle | Maple N. Chen Ricardo D. Santander Elijah J. Talamas Peter J. Jentsch Marie-Claude Bon Srđan G. Aćimović Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR Insects samurai wasp PCR identification species-specific primers <i>Halyomorpha halys</i> nondestructive DNA extraction |
| title | Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR |
| title_full | Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR |
| title_fullStr | Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR |
| title_full_unstemmed | Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR |
| title_short | Molecular Identification of <i>Trissolcus japonicus</i>, Parasitoid of the Brown Marmorated Stink Bug, by Species-Specific PCR |
| title_sort | molecular identification of i trissolcus japonicus i parasitoid of the brown marmorated stink bug by species specific pcr |
| topic | samurai wasp PCR identification species-specific primers <i>Halyomorpha halys</i> nondestructive DNA extraction |
| url | https://www.mdpi.com/2075-4450/12/5/467 |
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