LAMP based identification of phytoplasmas associated with cassava witches’ broom and sesame phyllody diseases in Vietnam
Quoc Nguyen: Nong Lam University
<div>Witches’ broom and phyllody symptoms have been caused by various groups of phytoplasmas resulting in severe yield losses of cassava and sesame respectively in Vietnam. Until now, many worldwide scientists have developed numerous techniques for phytoplasmas diagnostics, in particularly the use of nested PCR, RFLP, and sequencing. However, such methods are normally labor and time intensive and can only be taken in well-equipped labs. Therefore, an alternative method known as Loop Mediated Isothermal Amplification (LAMP) has been applied as a rapid, simple and low-cost diagnostic tool for phytoplasmas within one hour of sampling in the field that is appropriate to the poor and developing countries. In this study, the diagnostic procedures for cassava witches’ broom and sesame phyllody diseases were set up and validated with universal and specific LAMP primers of 16S rRNA and plant cytochrome oxidase (COX) primers used as amplification control. Furthermore, the reliability of LAMP assay was evaluated by using the nested PCR and phylogenetic analysis. We found the presence of phytoplasmas groups’ 16SrI, 16SrII in sesame and 16SrI, 16SrII, 16SrIV in cassava. The results from this study will be helpful for practical applications of LAMP-based diagnostics for the detection and control of different phytoplasmas associated diseases in plants.</div>
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