Molecular systematics and population genetics of Hypoxylon truncatum sensu Miller using RAPD
Yoon, Cheol-Sik
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https://hdl.handle.net/2142/21786
Description
Title
Molecular systematics and population genetics of Hypoxylon truncatum sensu Miller using RAPD
Author(s)
Yoon, Cheol-Sik
Issue Date
1992
Doctoral Committee Chair(s)
Glawe, Dean A.
Department of Study
Crop Sciences
Discipline
Crop Sciences
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Genetics
Biology, Microbiology
Agriculture, Plant Pathology
Language
eng
Abstract
Molecular systematics and population genetics of Hypoxylon truncatum sensu Miller were investigated, using random amplified polymorphic DNA markers (RAPD). A simple, rapid method was developed to isolate DNA from fungal tissue. This method described involves treating ground mycelium with a lysis buffer containing SDS, followed by precipitation of contaminants using CsCl. Using DNA isolated by this method, the effects of several parameters on PCR amplification were investigated. These parameters included different concentrations of DNA, MgCl$\sb2,$ dNTP, gelatin, primer, and Taq polymerase, different temperature profiles, and presence of RNA. The result of this study suggest that concentrations of reaction components studied are highly inter-dependant in their effects, and that RNA can interfere with PCR amplification of target DNA sequences.
A single population of H. truncatum from southern Illinois was studied. High degree of genetic variation was observed among single-ascospore isolates from a single stroma. Phenetically analyzed molecular data suggest that a clear association exists between RAPD markers and different stromatal types in the population studied. Another study investigated the genetic structure of six geographically distant populations. Phenetic analyses of a 99 character by 54 isolate matrix separated the 54 collections into two genetically distinct groups corresponding with different stromatal types. The results suggest that because the two stromatal forms of H. truncatum sensu Miller are genetically distinct, it would be useful to designate the different groups as different species. Phenogram branching patterns do not correspond with different populations studied, suggesting that gene flow likely occurs among these populations.
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