Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants
ABSTRACT Environmental DNA (eDNA) consists of genetic material shed by living organisms, including those that are deceased, offering a unique opportunity to detect and identify terrestrial insect pests without requiring visual identification. The sweetpotato whitefly, Bemisia argentifolii, and the t...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2024-09-01
|
| Series: | Environmental DNA |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/edn3.70026 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850200200396668928 |
|---|---|
| author | Jonathan Lee‐Rodriguez Christopher M. Ranger Ashley Leach Andrew Michel Michael E. Reding Luis Canas |
| author_facet | Jonathan Lee‐Rodriguez Christopher M. Ranger Ashley Leach Andrew Michel Michael E. Reding Luis Canas |
| author_sort | Jonathan Lee‐Rodriguez |
| collection | DOAJ |
| description | ABSTRACT Environmental DNA (eDNA) consists of genetic material shed by living organisms, including those that are deceased, offering a unique opportunity to detect and identify terrestrial insect pests without requiring visual identification. The sweetpotato whitefly, Bemisia argentifolii, and the twospotted spider mite, Tetranychus urticae, are notorious for causing crop losses through virus transmission and direct feeding. Our study aimed to: (1) assess the effectiveness of B. argentifolii literature‐based PCR primers compared to newly developed primers for eDNA amplification, (2) evaluate the sensitivity of conventional PCR (cPCR) and real‐time quantitative PCR (qPCR) for detecting eDNA of B. argentifolii and T. urticae, (3) establish a rapid eDNA processing methodology using the LGC Biosearch Technologies QuickExtract DNA extraction kit and the Qiagen DNeasy Blood and Tissue kit, and (4) test the specificity of the developed primers against non‐target species. B. argentifolii and T. urticae were confined to tomato leaves (Solanum lycopersicum) using clip cages for 24 h, after which eDNA was collected from leaf surfaces using a water spray method, filtered, and processed for DNA amplification. While literature‐based primers showed sufficient sensitivity, their specificity for eDNA applications was inadequate, prompting the design of novel PCR primers for both pest species. Positive eDNA detection was achieved with both amplification methods, with qPCR proving more reliable than cPCR due to the latter's inconsistent performance with positive control samples. We also introduced a rapid eDNA processing approach using the QuickExtract DNA extraction kit, contrasting it with the more conventional Qiagen DNeasy Blood and Tissue kit. We believe that our findings are the first step toward the practical use of eDNA as a highly sensitive, early detection technique. |
| format | Article |
| id | doaj-art-050da33e11d047179cae7d95bc632e19 |
| institution | OA Journals |
| issn | 2637-4943 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Environmental DNA |
| spelling | doaj-art-050da33e11d047179cae7d95bc632e192025-08-20T02:12:24ZengWileyEnvironmental DNA2637-49432024-09-0165n/an/a10.1002/edn3.70026Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato PlantsJonathan Lee‐Rodriguez0Christopher M. Ranger1Ashley Leach2Andrew Michel3Michael E. Reding4Luis Canas5Department of Entomology The Ohio State University Wooster Ohio USAUSDA‐Agricultural Research Service, Horticultural Insects Research Lab Wooster Ohio USADepartment of Entomology The Ohio State University Wooster Ohio USADepartment of Entomology The Ohio State University Wooster Ohio USAUSDA‐Agricultural Research Service, Horticultural Insects Research Lab Wooster Ohio USADepartment of Entomology The Ohio State University Wooster Ohio USAABSTRACT Environmental DNA (eDNA) consists of genetic material shed by living organisms, including those that are deceased, offering a unique opportunity to detect and identify terrestrial insect pests without requiring visual identification. The sweetpotato whitefly, Bemisia argentifolii, and the twospotted spider mite, Tetranychus urticae, are notorious for causing crop losses through virus transmission and direct feeding. Our study aimed to: (1) assess the effectiveness of B. argentifolii literature‐based PCR primers compared to newly developed primers for eDNA amplification, (2) evaluate the sensitivity of conventional PCR (cPCR) and real‐time quantitative PCR (qPCR) for detecting eDNA of B. argentifolii and T. urticae, (3) establish a rapid eDNA processing methodology using the LGC Biosearch Technologies QuickExtract DNA extraction kit and the Qiagen DNeasy Blood and Tissue kit, and (4) test the specificity of the developed primers against non‐target species. B. argentifolii and T. urticae were confined to tomato leaves (Solanum lycopersicum) using clip cages for 24 h, after which eDNA was collected from leaf surfaces using a water spray method, filtered, and processed for DNA amplification. While literature‐based primers showed sufficient sensitivity, their specificity for eDNA applications was inadequate, prompting the design of novel PCR primers for both pest species. Positive eDNA detection was achieved with both amplification methods, with qPCR proving more reliable than cPCR due to the latter's inconsistent performance with positive control samples. We also introduced a rapid eDNA processing approach using the QuickExtract DNA extraction kit, contrasting it with the more conventional Qiagen DNeasy Blood and Tissue kit. We believe that our findings are the first step toward the practical use of eDNA as a highly sensitive, early detection technique.https://doi.org/10.1002/edn3.70026DNAenvironmentalHemipteraInsectapolymerase chain reactionTetranychidae |
| spellingShingle | Jonathan Lee‐Rodriguez Christopher M. Ranger Ashley Leach Andrew Michel Michael E. Reding Luis Canas Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants Environmental DNA DNA environmental Hemiptera Insecta polymerase chain reaction Tetranychidae |
| title | Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants |
| title_full | Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants |
| title_fullStr | Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants |
| title_full_unstemmed | Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants |
| title_short | Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants |
| title_sort | using environmental dna to detect and identify sweetpotato whitefly bemisia argentifolii and twospotted spider mite tetranychus urticae in greenhouse grown tomato plants |
| topic | DNA environmental Hemiptera Insecta polymerase chain reaction Tetranychidae |
| url | https://doi.org/10.1002/edn3.70026 |
| work_keys_str_mv | AT jonathanleerodriguez usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants AT christophermranger usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants AT ashleyleach usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants AT andrewmichel usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants AT michaelereding usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants AT luiscanas usingenvironmentaldnatodetectandidentifysweetpotatowhiteflybemisiaargentifoliiandtwospottedspidermitetetranychusurticaeingreenhousegrowntomatoplants |