An efficient low-cost xylem sap isolation method for bacterial wilt assays in tomato
An efficient low-cost xylem sap isolation method for bacterial wilt assays in tomato
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DOI：
10.1002/aps3.11335
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发表时间：
2020-04-19
影响因子：
3.6
通讯作者：
Senthil-Kumar, Muthappa
中科院分区：
文献类型：
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作者：
Longchar, Bendangchuchang;Phukan, Tarinee;Senthil-Kumar, Muthappa
前言：建立了一种便携、简便、高效的番茄茎叶木质部汁液快速提取方法，用于番茄木质部定植青枯菌的诊断和定量检测。方法与结果利用手持无针注射器产生的负压从番茄茎段中提取木质部汁液。接种病原菌后4天，从不同土壤水分条件下生长的植株上采集样品。移液管尖端被改装为茎段的适配器。在Kelman的三苯基四氮唑(TTC)培养液中，通过电镀稀释液来定量提取的汁液中的细菌负荷。结论该方法简单、便携、可从预定大小的组织中提取样品，可在短时间内从大量样品中进行高通量现场采样，这是其他现有技术所无法实现的。
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.