Indexing peaches for RNA virus infection by isolation of replicative viral RNA /
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| Other Authors: | , , |
| Format: | Thesis Book |
| Language: | English |
| Published: |
1985.
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| Subjects: | |
| Online Access: | Link to OAKTrust copy Link to ProQuest copy |
| Abstract: | The most effective and economical means of controlling tree fruit viruses is through the use of virus-free propagation material and eradication. Indexing systems provide a means of identifying both virus-infected and virus-free trees. Peach viruses are currently indexed by grafting on indicator hosts. Usually these tests require weeks to months to complete. Serological and physical tests cannot be conducted with many peach viruses because of their inability to be mechanically isolated. For this research, an electrophoretic technique was adapted for isolating the replicative form of RNA viruses as a rapid and non-specific technique for indexing peach viruses. Peach leaf samples containing known viruses were obtained from different locations, 3 weeks after full bloom. Twenty gram specimens were frozen in liquid nitrogen, and held at -20 C until they were extracted. Both virus-free and virus-infected leaves were extracted with phenol. The dsRNA was isolated by one cycle of Whatman CF-11 cellulose chromatography, and analyzed on 5% polyacrylamide tube and/or slab gels. Resulting bands were visualized by ethidium bromide and Polaroid photography under ultra-violet light. Cucumber mosaic and tobacco mosaic viruses were used as positive controls and molecular weight markers. Virus-free peaches were used as negative controls. Once this technique was developed. TMV and CMV infected tissue consistently yielded dsRNA for 24 consecutive electrophoretic runs. The dsRNA structures for both controls and peach material were confirmed by resistance to RNase digestion in 0.3 M NaCl either before or after electrophoresis. No dsRNA bands were detected with the virus-free peaches. Double-stranded RNA was isolated from peaches containing Prunus necrotic ringspot, green ring mottle, or peach wart viruses. DsRNA was not detected in specimens infected with stem pitting, peach blotch, or apricot ring pox viruses. The results of this research indicate that this system can be used in combination with other techniques for rapidly indexing some peach viruses. |
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| Item Description: | Typescript (photocopy). Vita. "Major subject: Plant Pathology." |
| Physical Description: | vii, 99 leaves : illustrations ; 29 cm |
| Bibliography: | Includes bibliographical references (leaves 85-98). |