Division of Plant Protection, Central Potato Research Institute, Shimla-I.
Abstracts of Research Papers Presented during the National Symposium of Indian Virological Society at Unit of Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi-110 012, October 14–1.
A non-radioactive PSTVd detection protocol, based on the use of fluorescein labeled probe was standardized using Gene Images Random Prime Labeling Module (Amersham Pharmacia). Cloned PSTVd cDNA was restriction digested with Bam HI and labeled by random priming system. This protocol allows detection of hybrids (after a blocking step) by incubation with an antifluorescein alkaline phosphatse (AP) conjugate. The probe bound AP is used to catalyse light production by enzymatic decomposition of a substrate CDP-star. The signal (from dried leaf extract) was observed upto a dilution of 1: 256.
Further, a total of 125 in vitro tissue culture plantlets were also simultaneously tested using 32P labeled probe while screening with the fluorescein labeled probe. Similar results were obtained in both the cases, thereby proving the tests’ efficacy. This technique appears to have potential for a large-scale disease screening programme.
