Title : Simplified rapid membrane- based viral DNA isolation method for molecular diagnosis of graft transmissible pathogen in Citrus
Abstract:
Citrus is a widely distributed crop all over the world. Citrus is an important fruit crop in India. It is grown in about 0.50 million ha with an annual production of 0.39 million tones. Citrus is infected by a number of graft transmissible pathogen which include Citrus tristeza virus (CTV), Indian Citrus ring spot virus (IRSV), Citrus yellow mosaic virus (CYMV), Citrus exocortis viroid and the Citrus greening bacterium (Candidatus liberibacter asiaticus). These pathogens cause severe yield reduction in citrus and their detection in propagating material is an important component in bud- wood certification programme. Their reliable detection in infected citrus plants is now commonly done by PCR. A membrane based highly simplified DNA template preparation was standardized for detection of CBMV &Citrus greening. The Unified nucleic acid extraction protocol is non-phenol chloroform-based extraction protocol and was comparable with commercial DNA extraction kit for PCR detection of DNA pathogens in Citrus. The study has demonstrated that bark tissues of infected citrus plants can be common sources of nucleic acid template preparation for detection of this pathogens. Membrane based DNA template protocol is a novel approach in PCR detection of virus and greening bacterium and will be highly useful for survey, Sanitary and bud wood certification as well as for phytosanitary purpose in citrus.
A membrane based highly simplified DNA template preparation was standardized for detection of CBMV. The Unified nucleic acid extraction protocol is non-phenol chloroform-based extraction protocol and was comparable with commercial DNA extraction kit for PCR detection of DNA pathogens in Citrus
