Thesis
Detection of Erysiphe necator (Uncinula necator) with polymerase chain reaction and species-specific primers
Washington State University
Master of Science (MS), Washington State University
2003
Handle:
https://hdl.handle.net/2376/130
Abstract
A polymerase chain reaction (PCR) assay employing species-specific primers was developed to differentiate Erysiphe necator (Uncinula necator) from other powdery mildews common in the northwest United States. This assay is intended to be used in conjunction with high efficiency air samplers for the addition of an inoculum component to current grapevine powdery mildew risk assessment models. DNA was extracted from mycelia, conidia, and/or cleistothecia that were collected from grape leaves using a Burkard cyclone surface sampler. To differentiate E. necator from closely related powdery mildew fungi, primer pairs Uncin144 and Uncin511 were developed by aligning internal transcribed spacer (ITS) sequences from E. necator and other powdery mildews and choosing regions unique to E. necator. The primers generated amplicons specific to E. necator, but did not generate amplicons when tested with powdery mildew species collected from 46 disparate hosts in 26 vascular plant families. As a result of these tests, the amplification of a single 367 base pair (bp) fragment using the primer pairs, and visualized by gel electrophoresis, was considered evidence of the presence of E. necator. Amplification products were cloned and sequenced to verify the specificity of E. necator primers. This PCRbased test could enable the detection of E. necator in field samples within hours of collection, and when air sampling and identification protocols perfected, is expected to result in significant improvements to grapevine powdery mildew risk assessment models.
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Details
- Title
- Detection of Erysiphe necator (Uncinula necator) with polymerase chain reaction and species-specific primers
- Creators
- Jennifer Susan Falacy
- Contributors
- Gary G. Grove (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Plant Pathology, Department of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; Pullman, Wash. :
- Identifiers
- 99900525286301842
- Language
- English
- Resource Type
- Thesis