An efficient low-cost xylem sap isolation method for bacterial wilt assays in tomato
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Title |
An efficient low-cost xylem sap isolation method for bacterial wilt assays in tomato
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Creator |
Longchar, Bendangchuchang
Phukan, Tarinee Yadav, Sarita Senthil-Kumar, Muthappa |
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Subject |
negative pressure
Ralstonia solanacearum tomato stem xylem sap extraction |
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Description |
Accepted date: 12 February 2020.
PREMISE: A portable, simple, yet efficient method was developed for the rapid extraction of xylem sap from the stems and petioles of tomato plants for diagnostic and quantification assays of the xylem-colonizing wilt bacterium Ralstonia solanacearum. METHODS AND RESULTS: Xylem saps were extracted from tomato stem sections using negative pressure generated from handheld needleless syringes. The samples were collected from plants grown under different soil moisture levels at four days after inoculation with the pathogen. Pipette tips were modified to serve as adapters for the stem sections. The quantification of the bacterial load in the extracted sap was performed by plating sap dilutions in Kelman's triphenyltetrazolium chloride (TTC) medium. Pathogen identity was further confirmed by performing a PCR using R. solanacearum-specific primers. CONCLUSIONS: Due to its simplicity, portability, and thoroughness of extraction from predetermined tissue sizes, the method can potentially facilitate high-throughput onsite sampling from a large number of samples in a short time, which cannot be achieved with other available techniques. This work was supported by the Department of Science and Technology (DST), Government of India Science and Engineering Research Board (SERB) (SERB-National Postdoctoral Fellowship PDF/2017/002118 to B.L.) and by the Department of Biotechnology (project BT/09/IYBA/2015/07 to M.S.-K.). The authors thank Prof. S. K. Ray of Tezpur University, Assam, India, for providing the pathogen strains used in this study. The findings, opinions, and conclusions made in this study are explicitly those of the authors and do not necessarily reflect the views of the funding agencies. |
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Date |
2020-04-27T14:42:17Z
2020-04-27T14:42:17Z 2020 |
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Type |
Article
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Identifier |
Applications in Plant Sciences, 8(4): e11335
2168-0450 https://doi.org/10.1002/aps3.11335 https://bsapubs.onlinelibrary.wiley.com/doi/full/10.1002/aps3.11335 http://223.31.159.10:8080/jspui/handle/123456789/1056 |
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Language |
en_US
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Format |
application/pdf
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Publisher |
John Wiley & Sons
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