• Plant Biotechnology Reports
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• v.2 no.2
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• pp.145-152
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• 2008

Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

• Korean Journal of Oriental Medicine
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• v.8 no.1
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• pp.93-104
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• 2002

The following is a list of morphologic and genetic characteristics of Pinellia tuber. 1. The original plant of Pinellia tuber is Pinellia ternata$(T_{HUNB})$$B_{REIT}$. With regards to its external morphology, it is smaller than other Araceae species and its spadix is longer than its leaves, which trifurcate. 2. As regards its internal morphology, its mucous cell is elliptical and the vessel is helical or annular-shaped. Granules exist in abundance and in various shapes. 3. Distribution and size of laticifers are the key criteria on which to differentiate between domestic and imported Pinellia tuber. Laticifers are mainly distributed in the epidermis in domestic Pinellia tuber and in the cortical parenchyma in imported Pinellia tuber. The size of laticifers is somewhere between 1,3 and $8{\mu}m$ in diameter in imported Pinellia tuber bigger than its domestic counterpart. 4. RAPD markers display a great similarity in bands between domestic and Chinese Pinellia tuber. However, RAPD primers 352, 358, 365, 368 and 374 are distinctive markers for domestic Pinellia tuber. In the meantime, North Korean Pinellia tuber, morphologically similar to domestic Pinellia tuber, is genertically distinctive from its domestic counterpart in primers 354, 358, 365, 368, 374 and 379, a finding that supports the postulation that North Korean Pinellia tuber is tuber of another Araceae species.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.912-923
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• 2016

Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.

• The Korean Journal of Mycology
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• v.24 no.2 s.77
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• pp.111-126
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• 1996

Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

• Journal of Life Science
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• v.20 no.7
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• pp.983-990
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• 2010

Genus Spiraea is a woody species primarily distributed throughout Asia. Many species of this genus are important plants medicinally and ecologically. I evaluated a representative sample of the sixteen taxa with random amplified polymorphic DNA (RAPD) markers to estimate genetic relationships within genus Spiraea. In addition, RAPD analysis was also conducted to estimate the genetic diversity and population structure of these species. As the typical populations of Spiraea were small, isolated, and patchily distributed for natural populations, they maintained a low level of genetic diversity for polymorphic primers. The mean H was 0.117 across species. The Korean endemic species (S. chartacea) and patchily distributed species (S. betulifolia) showed fewer alleles per locus (mean 1.240 vs. 1.297), lower percent polymorphic locus (24.0 vs. 29.7), and lower diversity (0.092 vs. 0.121) than a relatively widely spread species. An assessment of the proportion of diversity present within species, $H_{POP}/H_{SP}$, indicated that about 87.8% the total genetic diversity was among species. Thus, the majority of genetic variation (87.8%) resided within species. The phylogenic tree showed three distinct groups. One clade includes S. prunifolia for. simpliciflora, S. thunbergii, S. chamaedryfolia var. ulmifolia, S. media, and S. cantoniensis. Another clade includes S. blumei, S. pubescens, S. chartacea, and S. chinensis. The other clade is the remaining seven species.

• The Korean Journal of Mycology
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• v.38 no.1
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• pp.34-39
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• 2010

We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

• Molecules and Cells
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• v.26 no.2
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• pp.146-151
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• 2008

The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

• Asian Journal of Turfgrass Science
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• v.24 no.2
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• pp.88-96
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• 2010

Zoysiagrass, especially Jungji implicating medium-leaf type zoysiagrass has been widely used in South Korea recently. This study was carried out to classify 36 selected medium-leaf type zoysiagrass accessions compared to 5 basic zoysiagrass species using RAPDs. Morphological characteristics such as leaf width, leaf angle, leaf sheath length, existance of trichomes and stolon color were measured as useful characteristics for identification of species in Zoysia genus. Nineteen RAPD markers were identified using 8 selected random primers. The dissimilarity coefficient of variants ranged from 0 to 0.736. Three zoysiagrass groups were clustered by dissimilarity coefficient analysis. Group 1 consisted of Z. japonica and some US varieties including 'Zenith' and 'Meyer'. Group 2 consisted of Z. sinica, Z. macrostachya and Korean commercial varieties such as 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type grasses. Group 3 was genetically distinct from Group 1 and Group 2, and included Z. matrella and Z. tenuifolia. 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type zoysiagrasses showed very close genetic relationship with Z. sinica and Z. macrostachya.

• Journal of Forest and Environmental Science
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• v.31 no.3
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• pp.214-224
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• 2015

The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

• Proceedings of the Ginseng society Conference
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• 1993.09a
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• pp.138-142
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• 1993

Korean ginseng has been widely used as medicine from ancient times in Asia. Current breeding efforts in Korea include the individual plant selection and the subsequent pure - line isolation, and considerable number of lines with desirable traits have thus been isolated. However, there were rare data on genetic maker and its analysis for selection of superior varieties. For taxonomic characterization and development of genetic markers for ginseng breeding, molecular biological methods including the RFLP and RAPD methods were applied. Cytoplasmic DNA of ginseng was analyzed for RFLP analysis. However. there is no different pattern among the chloroplast DNA or mitochondrial DNA of variants. In the case of RAPD analysis, the band patterns using 4 of 10 RAPD primers show the distinctive polymorphism among 9 ginseng variants, and lines, and Similarity Index(SI) on polymorphism was calculated for the extent and nature of these variabilities in ginseng. The sequences of 4 selected primers were TGCCGAGCTG, AATCGGGCTG. GAAACGGGTG, and GTGACGTAGG. By SI based on the polymorphic band patterns, Chungkyung - Chong and Hwangskoog - Chong, and JakyungChong 81783 and Jinjakyung of Russia showed the most close SI. The data of KG10l coincided with the fact that it was released from Hwangskoog - Chong. and Jakyung - Chong 81783 and Jinjakyung of Russia showed the most close SI. The data of KG101 coincided with the fact that it was released from Hwangskoog - Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.