• Horticultural Science & Technology
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• v.31 no.3
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• pp.337-343
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• 2013

The aims of this study were to compare genetic distances among 14 Phalaenopsis varieties using simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) marker systems and to determine the discrimination using SSR. A total of 111 SSR primers and 30 SRAP combinations were initially screened. Twelve SSR primers and thirty SRAP combinations showed high polymorphism among the 14 Phalaenopsis varieties including domestic breeding varieties, conserved in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected by silver staining method. A total of 474 polymorphic bands, including 55 by SSRs and 419 by SRAPs, were identified and used for genetic diversity analysis. Polymorphic bands were scored for calculating a simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. Fourteen Phalaenopsis varieties were classified into three major groups at similarity coefficient value of 0.683 and 0.66 using SRAP and SSR, respectively. Also we could discriminate these domestic breeding Palaenopsis varieties using only SSR 20 and SSR 22. The results indicate that SSR analysis is effective for discrimination among Phalaenopsis varieties and SRAP is useful for genetic diversity when there is no sequence information. These studied SSR and SRAP markers will be useful tools for genotype identification, germplasm conservation and genetic relationship study in Phalaenopsis.

• Korean Journal of Breeding Science
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• v.43 no.5
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• pp.399-404
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• 2011

Genetic diversity among 28 Cymbidium varieties was evaluated by using a sequence-related amplified polymorphism (SRAP) marker system. The SRAP marker which was based on the open reading frames (ORFs) regions was developed primarily for Brassica species, but has been applied to various crops. A total of 30 SRAP primer combinations were initially screened. Twenty-eight SRAP primer combinations showed high polymorphism among the 28 Cymbidium varieties, which were consisted of breeding varieties and their parents in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected silver staining method. One hundred ninety six polymorphic bands (7 per primer) were generated and ranged from 0.3 to 1.0 kb in size. Polymorphic fragments were scored for calculating simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. The mean genetic similarity coefficient value was 0.588. The results showed that the correlation between $F_1$ varieties and their parents was high. These studied SRAP markers will be useful tools for genotype identification, germplasm conservation, genetic relationships in Cymbidium.

• Korean Journal of Medicinal Crop Science
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• v.18 no.3
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• pp.180-185
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• 2010

We investigated genetic diversity among and within the populations of cultivated ginseng (Panax ginseng C. A. Meyer ) using SRAP profiles. A total of 24 ginseng plants were sampled from the three populations (two from China, one from Korea). Since all these populations are previously shown closely related to each other assister groups, we used Panax quinquefolium L. and wild ginseng as a reference species, which is not "within the sister group". All individuals from the three populations were screened with a total of 36 primer pairs with 26 primers generated from 328 SRAP bands of DNA gels. The mean gene diversity ($H_E$) was estimated to be 0.057 within populations (range 0.032-0.067), and 0.086 at the species level. The genetic differentiation (Gst=0.31) indicates that genetic variation apportioned 30% among populations and 70% within populations. Generally, the result of this study indicates that ginseng contains high molecular variation in its populations.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.303-311
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• 2017

In this study, random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) analyses were used for evaluation of genetic diversity of 61 kiwifruit (Actinidia spp.) germplasms including domestic and overseas collection cultivars. Forty RAPD primers were detected in a total of 230 polymorphic bands with an average of 5.75. Thirty-two SRAP primer combinations were detected in a total of 204 polymorphic bands with an average 6.38. By unweighted pair-group method arithmetic average cluster analysis using 434 polymorphic bands, kiwifruit germplasms were classified in three groups with similarity value of 0.680. Cluster I consisted of 46 kiwifruit germplasms belonging to A. deliciosa, A. chinensis, A. deliciosa ${\times}$ A. arguta, A. chinensis ${\times}$ A. arguta, and A. chinensis ${\times}$ A. deliciosa. Cluster II consisted of seven germplasms belonging to A. arguta and 'Skinny Green', a cultivar derived from a cross between A. arguta and A. deliciosa. Cluster III consisted of seven germplasms belonging to A. rufa, A. hemsleyana, A. macrosperma, A. polygama, and A. eriantha. Genetic similarity values among tested kiwifruit germplasms ranged from 0.479-0.991, and average similarity value was 0.717. Similarity value was highest (0.991) between NHK0038 (A. deliciosa) and NHK0040 (A. deliciosa), and lowest (0.479) between 'Hayward' (A. deliciosa) and K5-1-22 (A. arguta).

• Journal of Life Science
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• v.21 no.4
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• pp.502-508
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• 2011

The purpose of this study was to evaluate the methods of identifying zygotic seedlings of crosses between 'Morita unshiu' (Citrus. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco). In order to investigate the frequency and position of zygotic seedlings and to determine the efficiency of zygotic seedling identification, random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) were performed using UBC (9, 27, 229, 230, and 254) primers and F4/R27, F7/R14, F12/R10, and F44/R62 primer sets, respectively. A total of 37 putative zygotic seedlings out of 55 individuals were selected by RAPD and SRAP. The F7/R14 primer pair showed a screening ability of 45.5% (25/55), whereas the primer UBC27 revealed the highest efficiency of zygotic seedling identification (50.9%, 28/55). When both UBC27 and F7/R14 were properly used for selection of hybridized seedlings of 'Morita unshiu' (C. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco), screening efficiency was increased to 60% (33/55) for putative zygotic seedlings. Thus, it is possible to select putative hybrid zygotic seedlings in an accurate and effective manner by RAPD and SRAP.

• Korean Journal of Breeding Science
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• v.40 no.4
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• pp.401-407
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• 2008

This study was conducted to develop the DNA markers for identification of the strawberry cultivars in Korea and Japan. We developed fifteen cleaved amplified polymorphic sequence (CAPS) markers based on the Fragaria gene sequences. Among them six CAPS markers showed polymorphism exclusively in one cultivar. Five CAPS markers (ANR-MspI, ANR-BamHI, ACO-HinfI, DFR-AseI, FGT-MspI) provided enough polymorphism to identify eight Korean strawberry cultivars except for 'Maehyang' and 'Sunhong'. To complement the fifteen CAPS markers, we selected another fifteen sequence-related amplified polymorphism (SRAP) and one of them, me1/em5_460bp marker, made it possible to discriminate between 'Maehyang' and 'Sunhong'. Therefore, application of the five CAPS markers and one SRAP marker were sufficient to identify the nineteen Korean and Japanese strawberry cultivars. These markers could be used practically for cultivar identification of Korean and Japanese strawberry.

• Journal of Life Science
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• v.21 no.12
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• pp.1659-1665
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• 2011

In Citrus, an $F_1$ segregation population of 150 plants was constructed from a cross between 'Kiyomi' (C. unshiu ${\times}$ C. sinensis) carrying the male sterility trait and 'Jinkyool' (C. sunki). Sequence-related amplification polymorphism (SRAP) combined with bulked segregant analysis was used to develop markers linked to male fertility. In the $F_1$ population, 66 out of 150 seedlings had aborted anthers and the ratio of male sterile plants to fertile plants in the progenies matched the expected Mendelian segregation ratio of 1:1 ($x^2$ =2.16 at p=0.05). From the profiling of the 197 SRAP primer sets, three SRAP primer sets (F4/R27, F39/R60, and F15/R37) that were closely linked to the target trait were identified and successfully converted into a sequence characterized amplified region (SCAR) marker for selection of male fertility in citrus. The SCAR marker, using the pMS 33U/pMS 1462L primer set specifically, produced a single 1.4-Kb fragment that was linked to male fertility. Our results suggested that this SCAR marker can be useful for marker-assisted selection of male sterile individuals in breeding $F_1$ progenies in Citrus.

• Parasites, Hosts and Diseases
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• v.55 no.1
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• pp.31-37
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• 2017

This study aimed to investigate the morphology and reconstruct the phylogenetic relationships of Centrocestus formosanus originating from 5 species of freshwater fish, i.e., Esomus metallicus, Puntius brevis, Anabas testudineus, Parambassis siamensis, and Carassius auratus, in Chiang Mai province, Thailand. Sequence-related amplified polymorphism (SRAP) and phylogeny based on internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (CO1) were performed. The results showed similar morphologies of adult C. formosanus from day 5 after infection in chicks. C. formosanus originated from 4 species of freshwater fish had the same number of circumoral spines on the oral sucker, except for those from C. auratus which revealed 34 circumoral spines. The phylogenetic tree obtained from SRAP profile and the combination of ITS2 and CO1 sequence showed similar results that were correlated with the number of circumoral spines in adult worms. Genetic variability of C. formosanus also occurred in different species of freshwater fish hosts. However, more details of adult worm morphologies and more sensitive genetic markers are needed to confirm the species validity of C. formosanus with 34 circumoral spines originating from C. auratus in the future.

• The Plant Pathology Journal
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• v.32 no.2
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• pp.145-156
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• 2016

Fusarium graminearum Schwabe causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss of wheat and other cereal crops. Twelve isolates of F. graminearum were collected from naturally infected spikes of wheat from Assiut Egypt. These isolates were compared using SRAP. The results indicated distinct genetic groups exist within F. graminearum, and demonstrated that these groups have different biological properties, especially with respect to their pathogenicity on wheat. There were biologically significant differences between the groups; with group (B) isolates being more aggressive towards wheat than groups (A) and (C). Furthermore, Trichoderma harzianum (Rifai) and Bacillus subtilis (Ehrenberg) which isolated from wheat kernels were screened for antagonistic activity against F. graminearum. They significantly reduced the growth of F. graminearum colonies in culture. In order to gain insight into biological control effect in situ, highly antagonistic isolates of T. harzianum and B. subtilis were selected, based on their in vitro effectiveness, for greenhouse test. It was revealed that T. harzianum and B. subtilis significantly reduced FHB severity. The obtained results indicated that T. harzianum and B. subtilis are very effective biocontrol agents that offer potential benefit in FHB and should be harnessed for further biocontrol applications. The accurate analysis of genetic variation and studies of population structures have significant implications for understanding the genetic traits and disease control programs in wheat. This is the first known report of the distribution and genetic variation of F. graminearum on wheat spikes in Assiut Egypt.

• 한국균학회소식:학술대회논문집
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• 2009.10a
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• pp.3-13
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• 2009

90 wild Agaricus strains from China, including 44 Agaricus bisporus strains identified preliminarily by isozyme electrophoresis, were studied by the techniques of SRAP and ISSR. 18 special SRAP bands and 12 special ISSR bands were analyzed, the strains were clustered and a demdrogram was obtained. The results showed that the strains were divided into 2 groups, wild A. bisporus group and the other Agaricus group. It is similar to the result of isozyme electrophoresis. 41 wild A. bisporus strains from Sichuan and Tibet were divided into 4 groups based on their growing places, suggesting the regionally difference of the strains to be quite obvious. Some white wild A. bisporus strains from Xinjiang and Tibet had special patterns, resulting in lower coefficient values with other wild A. bisporus strains. The biological characteristics of three wild A. bisporus strains were analyzed, and the results showed: 1. The wild strains grew slowly on PDA medium with weak appressed mycelia, and grew normally in kernel or fermented cottonseed shell substrate. 2. They grew faster than control strain As2796 under lower temperature of $16^{\circ}C$, and higher temperature of $32^{\circ}C$, with optimum growing temperature of $20-24^{\circ}C$, which was $4^{\circ}C$ lower than that of control strain. 3. In the cultivation with manure compost via twice fermentation, the mycelia grew normally in compost and quite slowly in casing soil, and the fruitbodies occurred less and late with easily opening and low production. 4. The fruitbody was off-white with flat and scaled cap, long stipe and dark gill. The bisporus basidia occupied 70-80% and trisporus basidia 20-30% of the total basidia. 5. Heterokaryotic monospore isolates could fruit in cultivation, and the homokaryotic isolates could cross with those derived from overseas wild A.bisporus strains. 6. The electrophoresis phenotype of isozymes such as esterase etc. belonged to high production type (H type). 7. The RAPD patterns made much difference from those of high production, good quality or hybrid strains, which indicated that the wild strains produce a new kind of RAPD type.