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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 Xiu, L -
item Guo, J -
item Chen, R -
item Que, Y -
item Grisham, Michael

Submitted to: BioMed Research International
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 4, 2013
Publication Date: May 1, 2013
Citation: Liu, J., Xiu, 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. BioMed Research International. Volume 2013, Article ID 357692, 8 pages. http://dx.doi.org/10.1155/2013/357692.

Interpretive Summary: Ratoon stunt is a disease of sugarcane caused by the bacterium, Leifsonia xyli subsp. xyli (Lxx). It found in most sugarcane-planting countries and, if not controlled, can cause yield losses as high as 50%. The disease causes slower growth resulting in reduced yield. A visual inspection of a sugarcane plant cannot be used to determine if it is infected by the Lxx bacterium, because the disease does not cause characteristic external symptoms such as spots or lesions. A laboratory-based method is needed to determine if a plant is diseased. Most laboratory tests are either expensive, time consuming, or lack specificity or sensitivity. We developed a method for detecting the Lxx bacterium based on procedure with the acronym LAMP. The LAMP method provides a rapid, accurate, low-cost detection method for detecting Lxx in plant tissue without the need for specialized, expensive equipment. LAMP will be a valuable tool to test cuttings of sugarcane used to plant new fields to insure that the new planting is free of the disease.

Technical Abstract: Ratoon stunt, caused by the xylem-limited coryneform bacterium Leifsonia xyli subsp. xyli (Lxx), is prevalent in most sugarcane-planting countries. Because the disease does not cause characteristic external symptoms, a laboratory-based technique is needed for accurate diagnosis. Based on loop-mediated isothermal amplification (LAMP), we developed a method for detecting Lxx. The major advantages of the LAMP method are visual judgment by colour, reduced time (60 min), and not needing a costly PCR apparatus and gel scanner. 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 detection. The LAMP assay is a highly specific, rapid and sensitive method for the diagnosis of ratoon stunt caused by Lxx in sugarcane.

Last Modified: 10/1/2014
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