• Title/Summary/Keyword: RPB2

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Detection of single-nucleotide polymorphism in RPB2 of Wolfiporia hoelen strains and assessment of its applicability for strain breeding (복령 균주의 RPB2 유전자 내 단일염기다형성 및 육종 활용성 분석)

  • Su Yeon, Kim;Mi-Jeong, Park;Seong Hwan, Kim;Kang-Hyeon, Ka
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.199-207
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    • 2022
  • The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

Effects of rumen-protected biotin on the growth performance, nitrogen utilization and blood parameters of yearling Liaoning cashmere doelings

  • Haiying Liu;Ying Lin;Xuhui Chen;Guiqin Yang
    • Animal Bioscience
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    • v.36 no.11
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    • pp.1685-1692
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    • 2023
  • Objective: This study was conducted to investigate the effects of rumen-protected biotin (RPB) on growth performance, nutrient digestibility, nitrogen utilization and plasma biochemical parameters of Liaoning cashmere goats during the cashmere fiber growing period. Methods: Sixteen 6-month-old Liaoning cashmere twin-doelings (24.8±1.20 kg) were allocated to 2 diet groups that were individually ad libitum fed 30% concentrate and 70% forage diet (dry matter [DM]) by a paired experimental design. Goats of the control group were fed the basal diet, while goats belonging to the RPB group were fed the basal diet with 10 mg RPB/d per animal. The duration of the experiment was 16 weeks with two 8-week periods. Digestibility was determined at weeks 7 and 15, and other measures were taken every four weeks. Results: Compared with the control group, the average daily gain of the RPB group increased by 10.94% (p<0.05), and the intake of neutral detergent fiber was increased (p = 0.045). There were some increasing tendencies for the intake of DM, acid detergent fiber and ether extract (p = 0.070, 0.088, and 0.070, respectively). The intake and digestibility of N tended to increase (p = 0.062 and 0.093, respectively), while the N fecal excretion percentage of N intake was decreased (p = 0.093) in the RPB compared with the control group. N retention tended to increase (p = 0.084) with the addition of adding RPB to the diet. Plasma total protein was increased (p = 0.037), whereas the urea-N concentration was decreased (p = 0.049) in the RPB diet group compared with the control diet group. The levels of propionyl-CoA carboxylase (p<0.001) and methylmalonyl-CoA (p = 0.013) were increased in the RPB group. Conclusion: Supplementation of rumen-protected biotin in the diet of cashmere goats can enhance the utilization of N and improve daily weight gain during cashmere fiber growing period.

Protection Motivation Theory and Rabies Protective Behaviors Among School Students in Chonburi Province, Thailand

  • Laorujisawat, Mayurin;Wattanaburanon, Aimutcha;Abdullakasim, Pajaree;Maharachpong, Nipa
    • Journal of Preventive Medicine and Public Health
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    • v.54 no.6
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    • pp.431-440
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    • 2021
  • Objectives: The aim of this study was to predict rabies protective behaviors (RPB) based on protection motivation theory (PMT) among fourth-grade students at schools in Chonburi Province, Thailand. Methods: This cross-sectional study was conducted from December 2020 to February 2021. A multistage sampling technique was used for sample selection. The questionnaire was divided into socio-demographic data and questions related to PMT and RPB. Descriptive statistical analysis was conducted using the EpiData program and inferential statistics, and the results were tested using the partial least squares model with a significance level of less than 5%. Results: In total, 287 subjects were included, of whom 62.4% were girls and 40.4% reported that YouTube was their favorite media platform. Most participants had good perceived vulnerability, response efficacy, and self efficacy levels related to rabies (43.9, 68.6, and 73.2%, respectively). However, 54.5% had only fair perceived severity levels related to rabies. Significant positive correlations were found between RPB and the PMT constructs related to rabies (β, 0.298; p<0.001), and the school variable (S4) was also a predictor of RPB (β, -0.228; p<0.001). Among the PMT constructs, self efficacy was the strongest predictor of RPB (β, 0.741; p<0.001). Conclusions: PMT is a useful framework for predicting RPB. Future RPB or prevention/protection intervention studies based on PMT should focus on improving self efficacy and response efficacy, with a particular focus on teaching students not to intervene with fighting animals. The most influential PMT constructs can be used for designing tools and implementing and evaluating future educational interventions to prevent rabies in children.

Development of Penicillium italicum-Specific Primers for Rapid Detection among Fungal Isolates in Citrus

  • Chen, Kai;Tian, Zhonghuan;Jiang, Fatang;Long, Chao-an
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.984-988
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    • 2019
  • Blue mold in citrus is caused by Penicillium italicum. In this study, the P. italicum-specific primers were developed for rapid detection based on the conserved genes RPB1 and RPB2 among Penicillium genomes. The two primer pairs RPB1-a and RPB1-b proved to be specific to detect P. italicum. The PCR assay among 39 fungal isolates and the colonial, pathogenic morphologies and molecular methods validated the specificity and reliability of these two primer pairs. This report provided a method and P. italicum-specific primers, which might greatly contribute to citrus postharvest industry.

Molecular Phylogenetic Analysis of Botrytis cinerea Occurring in Korea (우리나라에 발생하는 잿빛곰팡이병균 Botrytis cinerea의 분자계통학적 유연관계)

  • Back, Chang-Gi;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.138-143
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    • 2014
  • Several isolates were collected from apple, pepper, strawberry, cucumber and tomato having typical gray mold symptoms. All the isolates were identified as Botrytis cinerea by using morphological characteristics and PCR-RFLP method. It was difficult to analyze the phylogenetic relationship of these isolates by using ITS region, HSP60 and G3PDH because these genes were highly homologous in their nucleotide in inter-species of B. cinerea and intra-species of genus Botrytis. However, phylogenetic analysis using combined sequences (RPB2, HSP60 and G3PDH genes) clearly showed that all isolate of B. cinerea were different from Botrytis spp. Furthermore, it was also confirmed that strawberry isolate was distantly related to apple, pepper, cucumber and tomato isolates that were closely related to each other in nucleotide level.

Delimitation of Russula Subgenus Amoenula in Korea Using Three Molecular Markers

  • Park, Myung Soo;Fong, Jonathan J.;Lee, Hyun;Oh, Seung-Yoon;Jung, Paul Eunil;Min, Young Ju;Seok, Soon Ja;Lim, Young Woon
    • Mycobiology
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    • v.41 no.4
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    • pp.191-201
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    • 2013
  • Distinguishing individual Russula species has been difficult due to extensive phenotypic plasticity and obscure morphological and anatomical discontinuities. Due to highly similar macroscopic features, such as the presence of a red-cap, species identification within the Russula subgenus Amoenula is particularly difficult. Three species of the subgenus Amoneula have been reported in Korea. We used a combination of morphology and three molecular markers, the internal transcribed spacer (ITS), 28S nuclear ribosomal large subunit (LSU), and RNA polymerase II gene (RPB2), for identification and study of the genetic diversity of Russula subgenus Amoenula in Korea. We identified only two species in Korea (R. mariae and R. violeipes); these two species were indistinguishable according to morphology and LSU, but were found to be reciprocally monophyletic species using ITS and RPB2. The markers, ITS, LSU, and RPB2, have been tested in the past for use as DNA barcoding markers, and findings of our study suggest that ITS and RPB2 had the best performance for the Russula subgenus Amoneula.

Specific PCR Detection of Four Quarantine Fusarium Species in Korea

  • Hong, Sae-Yeon;Kang, Mi-Ran;Cho, Eun-Ji;Kim, Hee-Kyoung;Yun, Sung-Hwan
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.409-416
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    • 2010
  • Fusarium species, a large group of plant pathogens, potentially pose quarantine concerns worldwide. Here, we focus on the development of a method for detecting four Fusarium species in quarantined plants in Korea: F. solani f. sp. cucurbitae, F. stilboides, F. redolens, and F. semitectum var. majus. Species-specific primers were designed from the nucleotide sequences of either the translation elongation factor-1 alpha (TEF1) gene or RNA polymerase II subunit (RPB2) gene. Two different primer sets derived from TEF1, all specific to F. solani f. sp. cucurbitae, were able to differentiate the two races (1 and 2) of this species. A set of nested primers for each race was designed to confirm the PCR results. Similarly, two primer sets derived from RPB2 successfully amplified specific fragments from five F. stilboides isolates grouped within a single phylogenetic clade. A specific TEF1 primer set amplified a DNA fragment from only four of the 12 F. redolens strains examined, which were grouped within a single phylogenetic clade. All of the F. semitectum var. majus isolates could be specifically detected with a single RPB2 primer set. The specificity of the primer sets developed here was confirmed using a total of 130 Fusarium isolates.

Multigene Phylogeny and Morphology of Ophiocordyceps alboperitheciata sp. nov., A New Entomopathogenic Fungus Attacking Lepidopteran Larva from Yunnan, China

  • Fan, Qi;Wang, Yuan-Bing;Zhang, Guo-Dong;Tang, De-Xiang;Yu, Hong
    • Mycobiology
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    • v.49 no.2
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    • pp.133-141
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    • 2021
  • A new fungus, Ophiocordyceps alboperitheciata, parasitic on the larva of Noctuidae (Lepidoptera) was identified from a survey of entomopathogenic fungi in Kunming Wild Duck Forest Park, Yunnan Province, China. It can be primarily distinguished from relatives by its longer fertile parts, sterile tips, superficial perithecia, narrower asci, and smaller septa of ascospores. As revealed from phylogenetic analyses inferred from nrSSU, nrLSU, tef-1α, rpb1, and rpb2 sequence data, O. alboperitheciata belongs to the Hirsutella citriformis clade in the genus Ophiocordyceps of Ophiocordycipitaceae, and forms a separated clade from other related species. The uniqueness of the taxon is significantly evidenced by both molecular phylogeny and morphology. Furthermore, the interspecific relationships in the H. citriformis clade are discussed.

Characterization of Fusarium solani Causing Fusarium Root Rot of Lisianthus in Korea (꽃도라지 뿌리썩음병을 일으키는 Fusarium solani 의 특성)

  • Choi, Hyo-Won;Hong, Sung Kee;Lee, Young Kee;Kim, Jeomsoon;Lee, Jae Guem;Kim, Hyo Won;Kang, Eun Hye;Lee, Eun Hyeong
    • The Korean Journal of Mycology
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    • v.45 no.1
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    • pp.74-82
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    • 2017
  • Lisianthus (Eustoma grandiflorum) is a flowering ornamental plant used widely in Korea. In 2015, wilting, damping-off, stunting, and root rot symptoms were observed in lisianthus plants of Yeoju and Gimhae, Korea. Affected seedlings appeared yellow and showed poor development of root systems in the field and in nursery boxes. Furthermore, affected plants were yellow, stunted, and died at approximately 2-3 months after transplanting. Fusarium species were consistently isolated from the basal stems of diseased plants. Nine isolates were identified as Fusarium solani based on morphological characteristics. Macroconidia of isolates were relatively wide, straight-to-slightly curved, and microconidia formed in false heads on long monophialides. Abundant chlamydospores were produced at the middle or tips of hyphae. To confirm this identification, a molecular analysis of the translation elongation factor 1 alpha (TEF) and RNA polymerase II subunit (RPB2) genes was conducted. The sequences of TEF and RPB2 showed 99.2-99.9% and 98.0-98.1% similarity, respectively, to those of reference F. solani strains in NCBI GenBank. Pathogenicity was tested using root dipping inoculation of healthy lisianthus seedlings. Symptoms were observed within 7 days of inoculation only in inoculated plants. This is the first report of F. solani causing Fusarium root rot on lisianthus in Korea.

Mycological Characteristics of Botrytis cinerea Causing Gray Mold on Ginseng in Korea (인삼 잿빛곰팡이병균 Botrytis cinerea의 균학적 특성)

  • Cho, Hye-Sun;Jeon, Yong-Ho;Do, Gyung-Ran;Cho, Dae-Hui;Yu, Yun-Hyun
    • Journal of Ginseng Research
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    • v.32 no.1
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    • pp.26-32
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    • 2008
  • A total of 29 Botrytis were isolated from ginseng gray mold at 8 locations in Korea from June to July, 2004 and 2005. The causative agent of them was identified as Botrytis cinerea based on morphological characteristics of conidia and RPB2 gene sequence analysis. In inoculation experiments with sclerotia, one isolate was pathogenic to ginseng plant whereas two isolates were non-pathogenic. Scanning electron microscopy revealed that the sclerotia of pathogenic isolate has rough surface and that of non-pathogenic isolate very smooth surface. Optimum temperature and culture conditions for sclerotia production were $5{\sim}20^{\circ}C$ and darkness, respectively. The number of sclerotia was increased tenfold on media added with 0.5% old stem fragment of ginseng.