• Development and Reproduction
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• v.13 no.2
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• pp.97-103
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• 2009

Genomic DNA samples isolated from four geographical freshwater crab (Eriocheir sinensis) populations collected in the inland of the Korean Peninsula (Gunsan, Paju, and Nampo) and a Chinese site, were used for PCR amplification. Seven decamer primers generated 19 specific loci (19/243 loci, 7.81%) in the Gunsan population, 32 (32/215 loci, 14.88%) in the Paju population, 19 (19/231 loci, 8.23%) in the Nampo population and 62 (62/340 loci, 18.24%) in a Chinese population. The average 8.9 specific loci exhibited inter-individual-specific characteristics, thus revealing DNA polymorphisms in the Chinese population. The number of unique shared loci to each population and number of shared loci by the four populations were generated by molecular analysis using seven primers in four populations. 35 unique shared loci to each population, with an average of 5.0 per primer, were observed in the Gunsan population, and 50 loci, with an average of 7.1 per primer, were observed in the Chinese population. The hierarchical dendrogram indicates three main branches: cluster 1 (GUNSAN 01$\sim$GUNSAN 05, PAJU 06$\sim$PAJU 10 and NAMPO 11$\sim$NAMPO 15) and cluster 2 (CHINESE 16, 17, 18, 19 and 20). Conclusively individual no. 20 of the PAJU 10 freshwater crab was most distantly related to CHINESE no. 20 (genetic distance = 0.667). Taken together, these results demonstrate the potential of RAPD analysis to identify diagnostic markers for the identification of four freshwater crab populations.

• Mycobiology
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• v.41 no.2
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• pp.86-93
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• 2013

Sequence characterized amplified region (SCAR) markers are one of the most effective and accurate tools for microbial identification. In this study, we applied SCAR markers for the rapid and accurate detection of Phytophthora katsurae, the casual agent of chestnut ink disease in Korea. In this study, we developed seven SCAR markers specific to P. katsurae using random amplified polymorphic DNA (RAPD), and assessed the potential of the SCAR markers to serve as tools for identifying P. katsurae. Seven primer pairs (SOPC 1F/SOPC 1R, SOPC 1-1F/SOPC 1-1R, SOPC 3F/SOPC 3R, SOPC 4F/SOPC 4R, SOPC 4F/SOPC 4-1R, SOPD 9F/SOPD 9R, and SOPD 10F/SOPD 10R) from a sequence derived from RAPD fragments were designed for the analysis of the SCAR markers. To evaluate the specificity and sensitivity of the SCAR markers, the genomic DNA of P. katsurae was serially diluted 10-fold to final concentrations from 1 mg/mL to 1 pg/mL. The limit of detection using the SCAR markers ranged from $100{\mu}g/mL$ to 100 ng/mL. To identify the limit for detecting P. katsurae zoospores, each suspension of zoospores was serially diluted 10-fold to final concentrations from $10{\times}10^5$ to $10{\times}10^1$ zoospores/mL, and then extracted. The limit of detection by SCAR markers was approximately $10{\times}10^1$ zoospores/mL. PCR detection with SCAR markers was specific for P. katsurae, and did not produce any P. katsurae-specific PCR amplicons from 16 other Phytophthora species used as controls. This study shows that SCAR markers are a useful tool for the rapid and effective detection of P. katsurae.

• Journal of Mushroom
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• v.12 no.1
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• pp.58-62
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• 2014

To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-68 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2798-24 derived from ASI 2798 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2798-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.2'. The 'Beesan No.2' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803, ASI 2798 and Beesan No.1. Analysis of the genetic characteristics of the new cultivar 'Beesan No.2' showed a different DNA profile as that of the control strains, ASI 2803, ASI 2798 and 'Beesan No.1', when RAPD(Random Amplified Polymorphic DNA) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 123.3 g. And also the stipe was long. This new cultivar 'Beesan No.2' of Pleurotus ferulae was characterized white-like variety of King oyster mushroom in the color of stipe, that was long and low yield compared to that of other cultuvar 'Beesan No.1'. We therefore expect that this new strain will increase of export and substitute cultivar of King oyster mushroom.

• Journal of Mushroom
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• v.12 no.1
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• pp.52-57
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• 2014

To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-95 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2850-24 derived from ASI 2850 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2850-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.1'. The 'Beesan No.1' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803 and ASI 2850. Analysis of the genetic characteristics of the new cultivar 'Beesan No.1' showed a different DNA profile as that of the control strains, ASI 2803 and ASI 2850, when RAPD(Random Amplified Polymorphic DNA) primer URP6 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 245 g using demonstration farms. And also the stipe was thick and long. This new cultivar 'Beesan No.1' of Pleurotus ferulae was characterized by fast fruitbody formation, the stipe was thick and long and high quality yield compared to that of other cultuvars. We therefore expect that this new strain will increase of farmer's income by construction of stabilized production system.

• Journal of Mushroom
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• v.11 no.2
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• pp.77-81
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• 2013

To develop a new cultivar of king oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2824-21 derived from ASI 2824(Keunneutari No. 2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-53(G09-21-10 x ASI 2844-9) was shown the best cultural characteristics, selected to be a new cultivar and designated as 'Seolsong'. The 'Seolsong' was distinctly formed incompatibility line in the confrontation growth of parental strains Keunneutari No. 2, Aeryni 3 and ASI 2844. Analysis of the genetic characteristics of the new cultivar 'Seolsong' showed a different DNA profile as that of the control strains, Keunneutari No. 2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25{\sim}30^{\circ}C$ and pH 5~8, respectively. This new cultivar 'Seolsong' of fruiting body production per bottle was about $131.4{{\pm}}43.1$ g which is about 102% quantity compared to that of other cultivar Keunneutari No. 2. And also the stipe is thick and long, but the number of available stipe is few. Particularly, it was tolerant of high moisture above 90% during the growth period after primodia formation. We therefore expect that this new strain will save of labor and cost of cultivation by without culling work.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.9
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• pp.1204-1209
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• 2004

Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

• Horticultural Science & Technology
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• v.33 no.4
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• pp.598-604
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• 2015

This study was carried out to develop molecular techniques to allow the selection of zygotic seedlings in the early stage of the plant development. We identified 37 pollen-specific molecular markers from RAPD analysis and successfully used them for identification of the zygotic seedlings from various hybrid crosses. Three Satsuma mandarin cultivars ('Morita unshiu', 'Nangan 20' and 'Miyagawawase') were used as mother parents and seven cultivars ('Ponkan', 'Lee', 'Kinokuni', 'Shiranuhi', 'Tamnaneunbong', 'Shinyegam', and 'Sunburst' mandarins) served as pollen parents. PCR analysis showed that 2 primers could identify zygotic hybrid seedlings. Among them, an UBC-27 primer was used to identify the zygotic seedlings from hybrid crosses of "'Nangan 20' ${\times}$ 'Kinokuni'" mandarin, "'Nangan 20 ${\times}$ Ponkan'" mandarin and "'Miyagawawase ${\times}$ Sunburst'" tangerine. In total 29 out of 40 seedlings (73%), 9 out of 47 seedlings (19%), and 13 out of 45 (29%) were identified as zygotic seedlings, respectively. These results can show that the pollen-specific markers selected in this study can be used effectively for early identification of zygotic seedlings from Citrus hybrid crosses.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1053-1061
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• 2004

Of the 20 arbitrarily chosen primers, six oligonucleotides decamer primers were used on the basis of the number of the polymorphisms generated in catfish (Silurus asotus) from Yesan and bullhead (Pseudobagrus fulvidraco) from Dangjin in Korea. Six primers were used generating a total of 602 scorable bands in catfish and 195 in bullhead population, respectively, ranging in size of DNA fragments from less than approximately 100 to larger than 2,000 base pairs (bp). Six primers yielded 199 polymorphic fragments (33.1%) in catfish and 47 (24%) in bullhead, respectively. In the present study, a total of 328 common fragments (an average of 54.7 per primer) were observed in catfish population, whereas 84 (an average of 14.0 per primer) in bullhead. The total number of specific fragments in catfish and bullhead population were 76 and 64, respectively. In catfish population, random decamer, OPA-17 (GACCGCTTGT) generated the highest number of fragments (a total of 141) in comparison with other primers used, with an average of 11.8. The common bands in the molecular weight of 300 bp generated by random primer OPA-06 (GGTCCCTGAC) were present in every individuals in bullhead population. The major polymorphic bands in the molecular weight of 100 bp generated by OPA-17 were identified in lane 14, 15, 17, 18, 19 20 and 21, which were identifying species in bullhead population. The average bandsharing values (BS values) of all of the samples within catfish population ranged from 0.575 to 0.945, whereas 0.063-1.000 within bullhead population. The bandsharing value (index of similarity between individuals) between individual No. 5 and No. 9 showed the highest level within catfish population, whereas the bandsharing value between individual No. 1 and No. 2 showed the lowest level. The single linkage cluster analysis resulted from four primers, indicating four genetic groupings composed of group 1 (C1-C10, all of the catfish samples), group 2 (B11, B12, B13, B14, B16, B17, B18, B19), group 3 (B15) and group 4 (B20 and B21). The dendrogram reveals close relationships between individual identities within two species populations and individuals derived from the same ancestor, respectively. However, genetic distances between two species populations ranged from 0.124 to 0.333. The shortest genetic distance (0.042) displaying significant molecular differences was between individual No. 6 and No. 9 catfish population. The shortest genetic distance (0.033) displaying significant molecular differences also was between individual No. 18 and No. 19 in bullhead population. Reversely, the genetic distance of individual No. 20/21 among individuals in bullhead population was highest (0.333). This result showed that bullhead No. 20 and 21 were distinct from other individuals within bullhead population.

• Korean Journal of Ichthyology
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• v.18 no.4
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• pp.300-310
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• 2006

Genomic DNA isolated from three geographical gizzard-shad (Konosirus punctatus) populations in Seocheon (SC), Busan (BS) and Gochang (GC) collected in the West Sea and the southern sea, respectively, off the Korean Peninsula, were PCR-amplified repeatedly. Eight selected decamer and 20-mer primers generated a total of 713 loci in the SC population, 791 in the BS population, and 732 in the GC population, with a DNA fragment size ranging from 100 bp to 2,800 bp. We identified 50 unique loci for the SC population, 70 unique loci for the BS population and 130 for the GC population: 120 shared loci for the three populations. There were 108 specific loci (15.1%) for the SC population, 74 (9.4%) for the BS population, and 67 (9.2%) for the GC population. Eight primers also generated 48 polymorphic loci (6.7%) for the SC population, 26 (3.3%) for the BS population, and 16 (2.2%) for the GC population. The similarity matrix ranged from 0.756 to 0.936 for the SC population, from 0.800 to 0.938 for the BS population, and from 0.731 to 0.959 for the GC population. The dendrogram obtained by the eight primers indicates three genetic clusters: cluster 1 (SEOCHEON 01~SEOCHEON 10), cluster 2 (BUSAN 11~BUSAN 20 and GOCHANG 23~GOCHANG 24), and cluster 3 (GOCHANG 21, 22, 25, 26, 27, 28, 29 and 30). As stated above, some individuals of the GC population appear to belong in BS population. When seeing this result, it was thought with the fact that some individuals of 2 populations seem to come and go partially. Thus, RAPD-PCR analysis revealed a significant genetic distance between the three geographical gizzard-shad populations. Using various decamer and 20-mer primers, RAPD-PCR may be applied to identify specific/polymorphic markers that are particular to a species and geographic population, and to define genetic diversity, polymorphisms, and similarities among geographical gizzard-shad populations.

• Journal of Mushroom
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• v.15 no.2
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• pp.84-87
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• 2017

Although ear mushroom (Auricularia auricula-judae) is cultivated worldwide, there are a limited number of commercial cultivars in Korea. Recent increase in the import of ear mushroom from China threatens the domestic farming. The present study introduces a new thermotolerant cultivar of ear mushroom, designated 'Hyeonyu', developed by mating monokaryons obtained from JBAA11 and CAA1 strains. The optimal growth temperature for Hyeonyu was 26-36?. The periods of primordia formation and fruit-body growth were 27 and 15 days, respectively. The average mushroom yield per bag (1 kg) was 350 g. Random amplification of polymorphic DNA (RAPD) analysis with OPA1 and OPA7 primers identified polymorphic DNA bands between the control, the new variety Hyeonyu, and a Chinese variety.