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PCR Detection and Sequence Analysis of the rDNA ITS Regions of Rhizina undulata  

Lee, Sun Keun (Department of Forest Resources Protection, College of Forest and Environmental Sciences, Kangwon National University)
Lee, Jong Kyu (Department of Forest Resources Protection, College of Forest and Environmental Sciences, Kangwon National University)
Kim, Kyung Hee (Division of Forest Insect Pests and Disease, Korea Forest Research Institute)
Lee, Seung Kyu (Division of Forest Insect Pests and Disease, Korea Forest Research Institute)
Lee, Sang Yong (Department of Forest Resources Protection, College of Forest and Environmental Sciences, Kangwon National University)
Publication Information
Journal of Korean Society of Forest Science / v.96, no.4, 2007 , pp. 425-431 More about this Journal
Abstract
To investigate genetic diversity and PCR detection of Rhizina undulata, PCR detection and sequence analysis of rDNA ITS region of R. undulata in soil were analyzed and developed. The length of partial 18S rDNA from four R. undulata isolates were 1,375 nt. The sequence similarity of R. undulata isolates was 100%. The rDNA ITS regions of R. undulata isolates were 585 nt long. Nucleotide sequencing of the ITS regions showed that PDK-1, PTT-1 and PDJ-9 isolates had 100% sequence identity. But, PDS-5 isolate differed from the three isolates by two nucleotide substitution. R. undulata-specific primers designed by the sequence of ITS region were used in PCR detection of R. undulata. PCR products about 525 bp size, which is specific to R. undulata, were amplified from total DNAs of R. undulata isolates. To assay the sensitivity of PCR detection by R. undulata ITS-specific primer, purely cultured mycelial suspension of R. undulata was serially diluted and mixed with 100g of sterile sandy loam soil, respectively. And then, PCR products of total DNAs extracted from each mycelium-soil mixtures were analysed. The PCR protocol could detected up to 1ng mycelium of R. undulata within 100g of soil.
Keywords
Rhizina undulata; I8S rDNA; ITS specific primer;
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