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KMID : 1160220140420010046
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Mycobiology
2014 Volume.42 No. 1 p.46 ~ p.51
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Isolation of a Variant Strain of Pleurotus eryngii and the Development of Specific DNA Markers to Identify the Variant Strain
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Lee Hyun-Jun
Kim Sang-Woo
Ryu Jae-San
Lee Chang-Yun
Ro Hyeon-Su
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Abstract
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A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplifiedpolymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealeddifferences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, wasisolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of thedegenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggestingthat chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primerswere designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp productin both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of theP2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in thenormal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushroomsincluding 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.
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KEYWORD
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Chromosome instability, Degeneration, Random amplified polymorphic DNA, Sectoring
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