• Title/Summary/Keyword: Sequence characterized amplified region

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Variability of Stem-Base Infestation and Coexistence of Fusarium spp. Causing Crown Rot of Winter Wheat in Serbia

  • Jevtic, Radivoje;Stosic, Nemanja;Zupunski, Vesna;Lalosevic, Mirjana;Orbovic, Branka
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.553-563
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    • 2019
  • Investigations related with factors influencing root and crown rot are rare and mainly related to farming practice and soil management. The main objective of this study was to examine broader range of factors influencing stem-base infestation of winter wheat in the field conditions. The effect of spatial distribution of infected plants on disease index (DIs) assessments was also investigated. Analysis of factors influencing DIs of crown rot of wheat demonstrated significant influence of the growing seasons (P < 0.001) and extreme fluctuations in winter temperatures (P < 0.001). In addition to that, localities together with their interaction with the growing season also significantly influenced DIs (P < 0.001). Aggregation of infected plants influenced variability of DI estimations, and it was pointed out that more extensive investigation should be conducted on broad range of DI in order to establish sampling method giving uniform sampling precision. Fusarium graminearum was shown to be predominant Fusarium species in Serbia (72.6%) using sequence-characterized amplified region analysis. Interestingly F. oxysporum was isolated in higher frequencies (27.4%) than it was reported in the literature. Given that there were no reports on the diversity of Fusarium species causing crown rot of wheat in Serbia, this study presents first report on this important subject. It also indicated that more attention should be focused on combined effects of abiotic and biotic factors influencing stem-base infestation of winter wheat. This knowledge will contribute to better understanding of factors influencing root and crown rot of wheat which would ensure sustainable disease management in the future.

Sero-epidemiology and genetic characterization of swine influenza virus (돼지 인플루엔자 바이러스의 혈청학적 역학조사 및 유전학적 분석)

  • Lyoo, Young-soo;Kim, Lomi
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.53-63
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    • 1998
  • Total of 1085 swine sera (1996-1997) from nation-wide were tested for the presence of antibodies to influenza A virus. Fifty nine percent of the tested sera showed seropositive by HI test. Positive sera consisted of 24--- of H3, 15--- of H1, and 20--- of the sample had both antibodies, respectively. Sera collected from various region represented 7~27--- seropositivity to H1N1, 15~25--- to H3N2, respectively. Swine influenza field isolate from nasal swab was characterized antigenically and genetically to elucidate its relatedness with other known strains of influenza A virus. The study was focused on the HA gene which is related to pathogenecity and antigenic variability of the influenza virus. By RT-PCR using influenza A/H1N1 specific primers, influenza virus H1N1 specific DNA fragment was amplified from A/Swine/Iowa/15/30(H1N1), US field isolate but not in H3N2 strain. PCR products were sequenced by dideoxy chain termination method to determine nucleotide homology with other strains of influenza A virus. The US field isolate and A/Swine/Indiana/1726/88 strain had 97--- of nucleotide homology and 98--- of amino acid homology. Based on the results obtained from this experiment, the field isolate was genetically related to A/Swine/Indiana/1726/88 and had higher homology with A/Swine/Indiana/1726/88 than with classical swine influenza virus, A/Swine/Iowa/15/30. The field isolate had no amino acid changes at the antigenic site compare to that of the A/Swine/Indiana/1726/88. The proteolytic enzyme cleavage site between HA1 and HA2 had no alteration and the amino acid arginine was intact. There is no evidence has been found that the field isolate has genetic shift or genetic drift which might altered antigenic determinant.

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Application of SCAR markers to self-incompatibility genotyping in breeding lines of radish (Raphanus sativus L.)

  • Chung, Hee;Kim, Su;Park, HanYong;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.397-402
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    • 2009
  • Self-incompatibility (SI) prevents self-fertilization by inhibiting the pollen tube growth of self-pollen. Molecular analysis has revealed that the S locus comprises a number of genes, such as the S-locus glycoprotein (SLG), the S-locus receptor kinase (SRK), and SP11 (SCR). Although molecular markers related to those genes have been developed, a simple S-haplotype detecting method has not been reported due to the highly polymorphic and relatively small coding regions. In this study, the sequence characterized amplified region (SCAR) markers were used to establish an efficient radish genotyping method. We identified the S-haplotypes of 192 radish accessions using 19 different markers, which proved to be highly reliable. The accessions were assigned to 17 types of S-haplotypes, including 8 types of SRKs and 9 types of SLGs. Since the developed SCAR markers are based on their gene sequences, we could easily identify the S-haplotypes by a single specific band, with the highest frequencies detected for SLG 5, SRK 1, and SLG 1, in order. Among the tested markers, the SLG 1, SRK 1, and SRK 5 markers exhibited high reliability, compared to phenotypic results. Furthermore, we identified the seven types of unreported SLGs using SLG Class -I and -II specific markers. Although the developed SCAR markers still need to be improved for the genotyping of all S-haplotypes, these markers could be helpful for monitoring inbred lines, and for developing the MAS in radish breeding programs.

Development and Application of Weonhyeong Strain-specific SCAR Marker in Pleurotus ostreatus (느타리 버섯에서 원형 품종 특이 SCAR marker 개발)

  • Seo, Kyoung-In;Jang, Kab-Yeul;Yoo, Young-Bok;Park, Soon-Young;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.22-30
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    • 2011
  • Weonhyeong is one of important commercial strains. It has good characteristics of bundle formation, grey colored pilei and high productivity. We previously reported grouping of 70 strains of Pleurotus ostreatus in which one group contained 35 strains including Weonhyeong. Four strains in that group showed same profiles implicating no variety distinction for mushroom cultivation. Now we developed a specific marker for identification of Weonhyeong. Sequence Characterized Amplified Region (SCAR) marker was developed from the RAPD amplicon. SCAR marker 'S-OPO5' produced only one band specific to 2183, 2240, 2595 and 2725 strains showing similar banding patterns to Weonhyeong in RAPD-PCR results. The sequence of 'S-OPO5' marker was unknown when compared with the data in the Genbank using BLASTN. BLASTX results indicated that the marker showed significant alignment with the protein sequences in Tricholoma bakamatsutake reverse transcriptase. The results indicate that this new SCAR marker ('S-OPO5') will be valuable to distinguish the Weonhyeong similar strains from Pleurotus spp.

Distribution of Subgroups in Pseudomonas syringae pv. actinidiae Biovar 3 Strains Isolated from Korea (국내에서 분리된 Pseudomonas syringae pv. actinidiae biovar 3 균주들의 subgroup 분포)

  • Lee, Young Sun;Kim, Gyoung Hee;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.1
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    • pp.52-58
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    • 2021
  • Pseudomonas syringae pv. actinidiae, which causes bacterial canker in kiwifruit, is divided into five biovars (1, 2, 3, 5, 6) on the basis of genetic characteristics and toxin productivity. Among them, biovar 3 is responsible for the current global outbreak, and has been isolated in Korea since 2011. Biovar 3 strains isolated from Korea are subdivided into six genetically different lineages (subgroup I, IV, V, VI, VII, and VIII) based on random amplified polymorphic DNA (RAPD) analysis. In this work, the subgroup-specific sequence characterized amplified region (SCAR) primers were developed from sequenced differential RAPD bands. Distribution of the subgroups of the biovar 3 strains collected in Korea from 2011-2017 were examined using these subgroup-specific primer sets. Among the 54 strains tested, 35 strains (64.8%) belonged to subgroup V, 9 strains (16.7%) belonged to subgroup IV, 4 strains (7.4%) belonged to subgroup VI, 3 strains (5.6%) belonged to subgroup VII, 2 strains (3.7%) belonged to subgroup VIII, and 1 (1.9%) strain belonged to subgroup I. Strains belonging to subgroups IV, V, and VI were shown to be related to strains isolated from China, New Zealand, and Chile, respectively. The study revealed that the biovar 3 strains in Korea are genetically diverse and are estimated to have been introduced through pollen sourced from foreign countries.

Development of a SCAR Marker Linked to Male Fertility Traits in 'Jinkyool' (Citrus sunki) ('진귤' (Citrus sunki) 의 웅성가임 연관 SCAR 마커 개발)

  • Chae, Chi-Won;Dutt, Manjul;Yun, Su-Hyun;Park, Jae-Ho;Lee, Dong-Hoon
    • 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.

Development SCAR marker for the rapid authenticaton of Batryticatus Bombyx based on COI Sequences (COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 -)

  • Kim, Wook Jin;Yang, Sungyu;Noh, Pureum;Park, Inkyu;Choi, Goya;Song, Jun-Ho;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.34 no.5
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    • pp.13-20
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    • 2019
  • Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

Development of RAPD-SCAR Molecular Marker Related to Seed-hair Characteristic in Carrot (당근(Daucus carota var. sativa) 종자모 형질 관련 RAPD-SCAR 분자표지 개발)

  • Shim, Eun-Jo;Park, Sung-Kwan;Oh, Gyu-Dong;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.756-763
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    • 2013
  • Mechanical hair removal of carrot seed causes seed injuries and suppresses the germination in carrot cultivation. This study was performed to develop molecular markers for breeding high quality cultivars with short-hair seed. To meet this objective, random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) markers specifically linked to seed-hair characteristic were identified using CT-SMR 616 OP 389-1 line with short-haired seed and CT-SMR 616 OP 616-33 line with long-haired seed, bred by self-pollination for 6 years from 2008 to 2013, as parents. After seed hair lengths of these lines were analyzed using microscope, next generations were advanced and compared with the molecular markers polymorphism. From RAPD analysis using fixed lines in 2011, twelve RAPD primers showing polymorphic bands specific between the two lines were identified from 80 random primers. To develop RAPD-SACR marker, SCAR primers were designed based on sequence analysis of these specific RAPD bands and more than three combinations of primers were tested. As a result, it was found that the $SCA2_{1.2}$ amplified single polymorphic band from short-haired seed line. To confirm this result, $SCA2_{1.2}$ marker was retested by applying to the 2012 and 2013 progenies. Finally, it was concluded that the developed $SCA2_{1.2}$ marker distinguished short-haired line from long-haired seed line. Therefore, SCAR marker, $SCA2_{1.2}$ is expected to be utilized for breeding of the short-haired seed cultivars.

Discrimination of Korean Apple Cultivars Using Combination of RAPD-SCAR Markers (RAPD-SCAR 마커 조합을 이용한 국내 육성 사과 품종 판별)

  • Cho, Kang-Hee;Heo, Seong;Kim, Hyun-Ran;Kim, Jeong-Hee;Shin, Il-Sheob;Han, Sang-Eun;Kim, Se-Hee;Kim, Dae-Hyun
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.828-835
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    • 2010
  • Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.