Affordable and reliable plant sap-mediated template preparation for the detection of various phytopathogens by PCR assay

作者：Sahana A B; Nagaraja H; Maheshwar P K; Sreenivasa M Y; Prasad M N Nagendra; Adkar Purushothama C R^*

来源：Phytoparasitica, 2014, 42(4): 519-527.

DOI：10.1007/s12600-014-0389-2

• Ca. Liberibacter
• Citrus tristeza virus
• Phytoplasma
• RT-PCR
• Viroids

Plant pathogens like %26apos;Ca. Liberibacter%26apos;, phytoplasma, viruses and viroids cause diseases to almost all economically important plants. A simple and fast sap-mediated polymerase chain reaction (PCR) method for the detection of these pathogens infecting various plant hosts is described in the present study. Sap from selected plants was drawn aseptically on parafilm, from the mid-rib of young leaves. Depending on the type of host plant, sap was diluted to optimal concentration before PCR analysis. %26apos;Ca. Liberibacter%26apos;, Citrus tristeza virus (CTV) and Citrus exocortis viroid (CEVd) infecting citrus, and %26apos;Ca. Phytoplasma%26apos; infecting pepper and sandal trees were tested by sap-mediated PCR. The reliability of this procedure was evaluated by comparing the findings with previously described protocols. The sap-mediated nucleic acid template preparation for PCR assay is devoid of laborious nucleic acid extraction and expensive chemicals. Hence the present method is rapid, economical and so can be employed for diagnosis of large number of plant samples.

• 出版日期2014-9