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Assessment of Genetic Relationship among Date (Zizyphus jujuba) Cultivars Revealed by I-SSR Marker (I-SSR 표지자분석을 이용한 대추나무 품종간 유연관계 분석)

  • Nam, Jae-Ik;Kim, Young-Mi;Choi, Go-Eun;Lee, Gwi-Young;Park, Jae-In
    • Journal of Korean Society of Forest Science
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    • v.102 no.1
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    • pp.59-65
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    • 2013
  • The jujube is an important fruit tree species in Korea. Traditionally, classifications of jujube cultivars have been based on morphological characters; however, morphological identification can be problematic because morphological traits are affected by environmental conditions. Therefore, DNA markers are now being used for the rapid and accurate identification of plant species. Inter-simple sequence repeat (I-SSR) is one of the best DNA-based molecular marker techniques, which is useful for studying genetic relations and for the identification of closely related cultivars. In this study, 5 Korean jujube trees and 1 jujube tree imported from China were analyzed for 16 I-SSR primers. Amplification of the genomic DNA of jujube cultivars by using I-SSR analysis generated 100 bands, with an average of 6.25 bands per primer, of which 45 bands (45%) were polymorphic. The number of amplified fragments with I-SSR primers ranged from 2 to 13. The percentage of polymorphism ranged from 10% to 100%. I-SSR finger printing profiles showed that 'Boeun jujube' and 'Daeri jujube' had characteristic DNA patterns, indicating unequivocal cultivar identification at molecular level. According to the results of clustering analysis, the genetic similarity coefficient ranged from 0.68 to 0.92. 'Boeun jujube' and 'Daeri jujube' were divided into independent groups, and 'Bokjo jujube', 'Geumseong jujube', 'Wolchul jujube', and 'Mudeung jujube' were placed in the same group. Therefore, I-SSR markers are suitable for the discrimination of 'Boeun jujube' and 'Daeri jujube' cultivars.

Luteinizing hormone beta gene polymorphism and its effect on semen quality traits and luteinizing hormone concentrations in Murrah buffalo bulls

  • Reen, Jagish Kour;Kerekoppa, Ramesha;Deginal, Revanasiddu;Ahirwar, Maneesh Kumar;Kannegundla, Uday;Chandra, Satish;Palat, Divya;Das, Dayal Nitai;Kataktalware, Mukund Amritrao;Jeyakumar, Sakthivel;Isloor, Shri krishna
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1119-1126
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    • 2018
  • Objective: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta ($LH{\beta}$) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. Methods: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in $LH{\beta}$ gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of $LH{\beta}$ gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. Results: $LH{\beta}$ gene was found to be polymorphic. Total six SNPs were identified in $LH{\beta}$ gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. Conclusion: The observed association between SSCP variants of $LH{\beta}$ gene with semen quality parameters and LH concentrations indicated the possibilities of using $LH{\beta}$ as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.

Multiplex Simple Sequence Repeat (SSR) Markers Discriminating Pleurotus eryngii Cultivar (큰느타리(Pleurotus eryngii) 품종 판별을 위한 초위성체 유래 다중 표지 개발)

  • Im, Chak Han;Kim, Kyung-Hee;Je, Hee Jeong;Ali, Asjad;Kim, Min-Keun;Joung, Wan-Kyu;Lee, Sang Dae;Shin, HyunYeol;Ryu, Jae-San
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.159-164
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    • 2014
  • For development of a method for differentiation of Pleurotus eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotyping results. PCR using each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in the range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pairgroup method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into two clusters. Cluster I and II were comprised of four and eight cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivars and strains with 21 alleles and a PIC value of 0.9090. These results might be useful in providing an efficient method for the identification of P. eryngii cultivars with separate PCR reactions.

Genetic Variation in the Endemic Rare Tree Species, Juniperus chinensis var. sargentii HENRY (희귀(稀貴) 수종(樹種) 눈향나무 집단(集團)의 동위효소(同位酵素) 분석(分析)에 의한 유전변이(遺傳變異) 연구(硏究))

  • Yang Byeung-Hoon;Kwon Hae-Yun;Han Sang-Don
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.76-82
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    • 2006
  • Genetic variation of two Juniperus chinensis var. sargentii populations in Mt. Seorak and Mt. Halla was investigated by isozyme analysis at reproducible 11 loci of 7 isozyme systems(Aat-1, Aat-2, Gdh, Idh, Lap, Mdh-1, Mdh-2, Mdh-3, 6Pgd, Pgi-1, and Pgi-2), of which 7 loci were polymorphic. The levels of genetic diversity of two populations were A=2.2, $A_e=1.61,\;P_{95}=54.5,\;H_{o}=0.179,\;H_e=0.287$(Mt. Seorak population) and A=2.1, $A_e=1.48,\;P_{95}=63.6,\;H_{o}=0.270,\;H_e=0.250$(Mt. Halla population), respectively. These values were similar to and/or somewhat higher than those observed in other Korean native conifers. Moderately low degree of genetic differentiation was observed between 2 analyzed populations ($F_{ST}=0.039$). Heterozygosity of the population in Mt. Seorak was significantly lower than expected, and much high level of inbreeding coefficient(F=0.376) was observed. Considering the limited population size and distribution range of the population, the population seemed to be influenced by inbreeding and/or random genetic drift, Consequently, Mt. Seorak population should be considered to be a more important candidate for the conservation of J. chinensis var. sargentii.

Characteristics of Leaf Morphology and Genetic Variation of the Rare Woody Plant, Berchemia racemosa var. magna (희귀수종 먹넌출 엽의 형태적 특성과 유전변이)

  • Song, Jeong-Ho;Lim, Hyo-In;Jang, Kyung-Hwan;Han, Jingyu
    • Korean Journal of Plant Resources
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    • v.26 no.5
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    • pp.613-618
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    • 2013
  • This study was conducted to investigate the morphological characteristics of leaf and the genetic diversity of Berchemia racemosa var. magna which is only found in Anmyeon Island of South Korea. ANOVA test showed that there were significant differences among individuals within population in all 10 leaf characteristics. Average characteristics of 39 individuals were 11.8 cm in leaf length, 7.1 cm in leaf width, 1.67 in leaf index, 5.4 cm in upper 1/3 width, 6.2 cm in lower 1/3 width, 3.6 cm in petiole length, 0.19 mm in leaf thickness, 11.5 ea. in number of veins (left), 11.4 ea. in number of veins (right) and 61.7 $cm^2$ in leaf area, respectively. Except for leaf thickness (18.8%), petiole length (21.7%) and leaf area (22.0%), the coefficients of variation of most leaf characteristics were relatively low (<15.0%). A total of 50 bands was generated from 8 selected I-SSR primers. The estimates of genetic variation were 1.719 in effective number of alleles ($A_e$), 26.0% in proportion of polymorphic bands (P), 0.410 in expected heterozygosity ($H_e$) and 0.598 in Shannon's diversity index (S.I.), respectively. In spite of the small number and the limited distribution, the B. racemosa var. magna population in Anmyeon Island showed high genetic diversity.

Lack of allozyme variation in the two carnivorous, terrestrial herbs Utricularia bifida and Utricularia caerulea (Lentibulariaceae) co-occurring on wetlands in South Korea: Inference of population history (한반도 남부 지방 습지에 같이 자생하는 식충 육상 초본 2종 땅귀개 및 이삭귀개 (통발과)의 알로자임 변이의 결여: 집단의 역사 추론)

  • Chung, Mi Yoon;Lopez-Pujol, Jordi;Chung, Myong Gi
    • Korean Journal of Plant Taxonomy
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    • v.47 no.4
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    • pp.297-303
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    • 2017
  • In central and southern Korea, the two small insectivorous, terrestrial herbs, Utricularia bifida and U. caerulea, often co-occur at wet locations (or in wetlands). The Korean Peninsula (with central China and northern Japan) constitutes the northern edge of their distribution, as their main range is subtropical and tropical Asia. The Korean populations of both species are very likely of post-glacial origin, given that warm-temperate vegetation was absent from the Korean Peninsula during the Last Glacial Maximum. Two hypotheses of the post-glacial colonization of the peninsula can be formulated; first, if current populations were founded by propagules coming from a single ancestral population (i.e., a single refugium), we would expect low levels of genetic diversity. Alternatively, if contemporary Korean populations originated from multiple sources (multiple refugia), we would expect high levels of genetic variation. To test which is more likely, we surveyed the degree of allozyme variation at 20 loci in ten populations for each of the two species from southern Korea. We found no allozyme variation within each species. However, their aquatic congener U. australis exhibited allozyme polymorphism across Japan (four polymorphic loci at three enzyme systems). We suggest that southern Korean populations of Utricularia bifida and U. caerulea were established by a single introduction event from a genetically depauperate ancestral population.

Genetic diversity among cultivated and wild Panax ginseng populations revealed by high-resolution microsatellite markers

  • Jang, Woojong;Jang, Yeeun;Kim, Nam-Hoon;Waminal, Nomar Espinosa;Kim, Young Chang;Lee, Jung Woo;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.637-643
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    • 2020
  • Background: Ginseng (Panax ginseng Meyer) is one of the world's most valuable medicinal plants with numerous pharmacological effects. Ginseng has been cultivated from wild mountain ginseng collections for a few hundred years. However, the genetic diversity of cultivated and wild ginseng populations is not fully understood. Methods: We developed 92 polymorphic microsatellite markers based on whole-genome sequence data. We selected five markers that represent clear allele diversity for each of their corresponding loci to elucidate genetic diversity. These markers were applied to 147 individual plants, including cultivars, breeding lines, and wild populations in Korea and neighboring countries. Results: Most of the 92 markers displayed multiple-band patterns, resulting from genome duplication, which causes confusion in interpretation of their target locus. The five high-resolution markers revealed 3 to 8 alleles from each single locus. The proportion of heterozygosity (He) ranged from 0.027 to 0.190, with an average of 0.132, which is notably lower than that of previous studies. Polymorphism information content of the markers ranged from 0.199 to 0.701, with an average of 0.454. There was no statistically significant difference in genetic diversity between cultivated and wild ginseng groups, and they showed intermingled positioning in the phylogenetic relationship. Conclusion: Ginseng has a relatively high level of genetic diversity, and cultivated and wild groups have similar levels of genetic diversity. Collectively, our data demonstrate that current breeding populations have abundant genetic diversity for breeding of elite ginseng cultivars.

Intergeneric Transfer of Isolated Nuclei from Lentinus edodes into Protoplasts of Pleurotus florida (사철느타리버섯 원형질체내에 표고 핵의 전이)

  • Yoo, Young-Bok;Shin, Pyung-Gyun
    • 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.

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Morphological and Genetic Characteristics of Colletotrichum gloeosporioides Isolated from Newly Emerging Static-Symptom Anthracnose in Apple

  • Jeon, Yongho;Cheon, Wonsu
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.34-34
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    • 2014
  • Filamentous fungi of the genus Colletotrichum (teleomorph, Glomerella) are considered major plant pathogens worldwide. Cereals, legumes, vegetables, and fruit trees may be seriously affected by this pathogen (1). Colletotrichum species cause typical disease symptoms known as anthracnoses, characterized by sunken necrotic tissue, where orange conidial masses are produced. Anthracnose appears in both developing and mature plant tissues (2). We investigated disease occurrence in apple orchards from 2013 to 2014 in northern Gyeongbuk province, Korea. Typical anthracnose with advanced symptoms was observed in all apple orchards studied. Of late, static fruit spot symptoms are being observed in apple orchards. A small lesion, which does not expand further and remains static until the harvesting season, is observed at the beginning of fruit growth period. In our study, static symptoms, together with the typical symptoms, were observed on apples. The isolated fungus was tested for pathogenicity on cv. 'Fuji apple' (fully ripe fruits, unripe fruits, and cross-section of fruits) by inoculating the fruits with a conidial suspension ($10^5$ conidia/ml). In apple inoculated with typical anthracnose fungus, the anthracnose symptoms progressed, and dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. However, in apple inoculated with fungi causing static symptoms, the size of the spots did not increase. Interestingly, the shape and size of the conidia and the shape of the appressoria of both types of fungi were found to be similar. The conidia of the two types of fungi were straight and cylindrical, with an obtuse apex. The culture and morphological characteristics of the conidia were similar to those of C. gloeosporioides (5). The conidia of C. gloeosporioides germinate and form appressoria in response to chemical signals such as host surface wax and the fruitripening hormone ethylene (3). In this study, the spores started to germinate 4 h after incubation with an ethephon suspension. Then, the germ tubes began to swell, and subsequently, differentiation into appressoria with dark thick walls was completed by 8 h. In advanced symptoms, fungal spores of virtually all the appressoria formed primary hyphae within 16 h. However, in the static-symptom fungus spores, no primary hyphae formed by 16 h. The two types of isolates exhibited different growth rates on medium containing apple pectin, Na polypectate, or glucose as the sole carbon. Static-symptom fungi had a >10% reduction in growth (apple pectin, 14.9%; Na polypectate, 27.7%; glucose, 10.4%). The fungal isolates were also genetically characterized by sequencing. ITS regions of rDNA, chitin synthase 1 (CHS1), actin (ACT), and ${\beta}$-tubulin (${\beta}t$) were amplified from isolates using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, ACT-512F and ACT-783R, and T1 and ${\beta}t2$ (5), respectively. The resulting sequences showed 100% identity with sequences of C. gloeosporioides at KC493156, and the sequence of the ${\beta}$t gene showed 100% identity with C. gloeosporioides at JX009557.1. Therefore, sequence data from the four loci studied proves that the isolated pathogen is C. gloeosporioides. We also performed random amplified polymorphic DNA-PCR, which showed clearly differentiated subgroups of C. gloeosporioides genotypes. The clustering of these groups was highly related to the symptom types of the individual strains.

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Amplified fragment length polymorphism fingerprinting analysis of Staphylococcus aureus isolated from bovine mastitis milk (소 유방염 유래 Staphylococcus aureus의 AFLP 지문분석)

  • Kim, Yeon-soo;Kim, Sang-kyun;Hwang, Eui-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.157-165
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    • 2001
  • Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

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