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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
 
Creator Longchar, Bendangchuchang
Phukan, Tarinee
Yadav, Sarita
Senthil-Kumar, Muthappa
 
Subject negative pressure
Ralstonia solanacearum
tomato stem
xylem sap extraction
 
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.
 
Date 2020-04-27T14:42:17Z
2020-04-27T14:42:17Z
2020
 
Type Article
 
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
 
Language en_US
 
Format application/pdf
 
Publisher John Wiley & Sons