{"title":"Isolation and Analysis of Double-Stranded RNA from Virus-Infected Plant and Fungal Tissue.","authors":"T J Morris, J A Dodds","doi":"10.1094/Phyto-69-854","DOIUrl":null,"url":null,"abstract":"<p><p>A simple, rapid method for the isolation of double-stranded RNA (dsRNA) from virus-infected plant and fungal tissues provides a new approach to virus detection and identification. Diseased tissue was phenol-extracted to isolate cellular nucleic acids, and viral dsRNA was selectively purified from other nucleic acids by binding to cellulose powder in 15% ethanol either in small columns or by a batch procedure. The product was analyzed first by gel electrophoresis and then by ribonuclease treatment to identify dsRNA. The method permits rapid and efficient isolation and analysis of dsRNA from small amounts (1-10 g) of tissue and from multiple samples using small amounts (0.1-2.5 g) of cellulose powder. Successful isolation of dsRNA does not depend on the type of tissue processed and the method therefore is potentially useful for the study of RNA virus replication and for detection and diagnosis of virus infections directly from the infected host tissues.</p>","PeriodicalId":20410,"journal":{"name":"Phytopathology","volume":"69 1","pages":"854-858"},"PeriodicalIF":3.1000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Phytopathology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1094/Phyto-69-854","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
A simple, rapid method for the isolation of double-stranded RNA (dsRNA) from virus-infected plant and fungal tissues provides a new approach to virus detection and identification. Diseased tissue was phenol-extracted to isolate cellular nucleic acids, and viral dsRNA was selectively purified from other nucleic acids by binding to cellulose powder in 15% ethanol either in small columns or by a batch procedure. The product was analyzed first by gel electrophoresis and then by ribonuclease treatment to identify dsRNA. The method permits rapid and efficient isolation and analysis of dsRNA from small amounts (1-10 g) of tissue and from multiple samples using small amounts (0.1-2.5 g) of cellulose powder. Successful isolation of dsRNA does not depend on the type of tissue processed and the method therefore is potentially useful for the study of RNA virus replication and for detection and diagnosis of virus infections directly from the infected host tissues.
期刊介绍：
Phytopathology publishes articles on fundamental research that advances understanding of the nature of plant diseases, the agents that cause them, their spread, the losses they cause, and measures that can be used to control them. Phytopathology considers manuscripts covering all aspects of plant diseases including bacteriology, host-parasite biochemistry and cell biology, biological control, disease control and pest management, description of new pathogen species description of new pathogen species, ecology and population biology, epidemiology, disease etiology, host genetics and resistance, mycology, nematology, plant stress and abiotic disorders, postharvest pathology and mycotoxins, and virology. Papers dealing mainly with taxonomy, such as descriptions of new plant pathogen taxa are acceptable if they include plant disease research results such as pathogenicity, host range, etc. Taxonomic papers that focus on classification, identification, and nomenclature below the subspecies level may also be submitted to Phytopathology.
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