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United States Department of Agriculture

Agricultural Research Service

Research Project: Effective Disease Management Through Enhancement of Resistant Sugarcane

Location: Sugarcane Research Unit

Title: Development of loop-mediated isothermal amplification for detection of Leifsonia xyli subsp. xyli in sugarcane

Authors
item Liu, J -
item Xu, L -
item Guo, J -
item Chen, R -
item Que, Y -
item Grisham, Michael

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: March 18, 2013
Publication Date: June 1, 2013
Citation: Liu, J., Xu, L.P., Guo, J.L., Chen, R.K., Que, Y.X., Grisham, M.P. 2013. Development of loop-mediated isothermal amplification for detection of Leifsonia xyli subsp. xyli in sugarcane. Phytopathology. (Suppl.2):S2.84.

Technical Abstract: Ratoon stunt, caused by the xylem-limited coryneform bacterium Leifsonia xyli subsp. xyli (Lxx), is prevalent in most sugarcane-producing countries. Because the disease does not cause characteristic external symptoms, a laboratory-based technique is needed for accurate diagnosis. We developed a diagnostic protocol to detect Lxx in host tissue based on loop-mediated isothermal amplification (LAMP). The LAMP assay is a highly specific, rapid, and sensitive method for the diagnosis of ratoon stunt caused by Lxx in sugarcane. This is a simple and feasible diagnostic tool in which the reaction takes place in a single tube incubated in a heat block at a constant temperature for 62 min compared to conventional PCR that takes about 2 h to detect Lxx. Amplified products are detected by a visual color change. The LAMP assay also does not require a thermocycler or an electrophoresis system, costly instruments required for conventional PCR. In the present study, positive and negative samples detected by the LAMP method were clearly distinguishable. When total DNA extracted from internode juice was used as the template, the sensitivity of LAMP was 10 times higher than that of the conventional PCR analysis.

Last Modified: 9/20/2014
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