Development of a Loop-mediated Isothermal Amplification for the Detection of Burkholderia glumae
Semester of Graduation
Master of Science (MS)
Plant Pathology and Crop Physiology
Bacterial panicle blight is among the three most limiting diseases of rice production in the United States. Yield loss up to 40% has been reported from rice fields severely infested with bacterial panicle blight. The disease is caused by Burkholderia glumae and B. gladioli. Although, both species produce similar symptoms and are closely related, but B. glumae strains are more aggressive and cause more severe symptoms on rice. Bacterial panicle bight is difficult to manage in the absence of effective chemical control measures. Rapid and early disease detection is needed to avoid severe yield losses. Several techniques have been developed for bacterial identification, but these methods are time consuming and some require high-precision instruments for amplification and analysis of target DNA. We developed a Loop-Mediated Isothermal Amplification (LAMP) protocol for rapid detection of B. glumae using a set of six primers from the gyrB housekeeping gene. Several commercially available dyes including, PicoGreen, Hydroxynaphthol Blue (HNB) and Calcein were compared to analyze the LAMP product. The LAMP detection method resulted in rapid and accurate detection of B. glumae. Among the different detection dyes, PicoGreen and HNB produced reliable results in the detection of B. glumae. Although, both produced accurate results; however, HNB is more cost-effective.
Document Availability at the Time of Submission
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Caldera, Maria Alejandra, "Development of a Loop-mediated Isothermal Amplification for the Detection of Burkholderia glumae" (2014). LSU Master's Theses. 3384.