Double-stranded RNA isolated from grapevine affected by grapevine stem-pitting disease Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/th83m3633

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  • Nucleic acid extracted from dormant cane phloem of grapevines affected by grapevine stem-pitting disease (SP) formed a high molecular weight (MW) band on 1.0% and 0.8% agarose gels. The band was unique to diseased vines. Other bands appeared from samples of both diseased and virus-free vines, but did not associate consistently with either. The nucleic acid species associated with stem-pitting-diseased vines was between 5.4 and 6.2 x 10⁶ MW and consisted of at least some double-stranded RNA (dsRNA), as suggested by its resistance to digestion by DNase, and its green fluorescence under acridine-orange staining. Double-stranded RNA species from Geotrichum candidum (hypovirulent strain 223), Helminthosporium maydis (hypovirulent strain HM9), citrus infected with citrus tristeza virus(strain T505), and alfalfa mosaic virus-infected Ajuga reptans were used as size standards and high MW-bracketing markers. Stem-pitting-affected tissue samples were not recognized by grapevine virus "A" immunoglobulins, but the size of the SP-associated dsRNA suggests that it might represent the replicative form of an 800 nm SP-associated virus. The possibility of using dsRNA electrophoretic patterns as a quarantine screening technique is discussed.
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