• Title/Summary/Keyword: $\beta$-tubulin

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Aspergillus caninus (Syn: Phialosimplex caninus): a New Isolate from Field Soils in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Sang Woo;Lee, Hyun Goo;Ju, Han Jun;Gwon, Byeong Heon;Kosol, San;Bazie, Setu;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.383-392
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    • 2018
  • During the study of indigenous fungal communities in soil samples collected from various field soils in Sancheong, Gyeongsangnam-do, Korea in 2017, several species of Aspergillus were discovered. Aspergillus caninus (KNU17-7) was isolated, identified, and described based on the results from macro and micro morphological characteristics and molecular characterization. Morphologically, it was identified using five different growth media: potato dextrose agar, oatmeal agar, yeast extract sucrose agar, czapek yeast extract agar, and malt extract agar. For the molecular identification, sequencing of internal transcribed spacer, ${\beta}-tubulin$, and calmodulin genes was performed. Based on this characterization, our study isolate was identified as Aspergillus caninus. This fungal species has not been officially reported in Korea before, and we report here with its morphological and molecular phylogenetic characterization.

Genetic Diversity of the Pear Scab Fungus Venturia nashicola in Korea

  • Choi, Eu Ddeum;Kim, Gyoung Hee;Park, Sook-Young;Song, Jang Hoon;Lee, Young Sun;Jung, Jae Sung;Koh, Young Jin
    • Mycobiology
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    • v.47 no.1
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    • pp.76-86
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    • 2019
  • Scab disease caused by Venturia nashicola is of agroeconomic importance in cultivation of Asian pear. However, little is known about the degree of genetic diversity in the populations of this pathogen. In this study, we collected 55 isolates from pear scab lesions in 13 major cultivation areas in Korea and examined the diversity using sequences of internal transcribed spacer (ITS) region, ${\beta}$-tubulin (TUB2), and translation elongation factor-$1{\alpha}$ ($TEF-1{\alpha}$) genes as molecular markers. Despite a low level of overall sequence variation, we found three distinctive subgroups from phylogenetic analysis of combined ITS, TUB2, and $TEF-1{\alpha}$ sequences. Among the three subgroups, subgroup 1 (60% of isolates collected) was predominant compared to subgroup 2 (23.6%) or subgroup 3 (16.4%) and was distributed throughout Korea. To understand the genetic diversity among the subgroups, RAPD analysis was performed. The isolates yielded highly diverse amplicon patterns and none of the defined subgroups within the dendrogram were supported by bootstrap values greater than 30%. Moreover, there is no significant correlation between the geographical distribution and the subgroups defined by molecular phylogeny. Our data suggest a low level of genetic diversification among the populations of V. nashicola in Korea.

Ascospore Infection and Colletotrichum Species Causing Glomerella Leaf Spot of Apple in Uruguay

  • Alaniz, Sandra;Cuozzo, Vanessa;Martinez, Valentina;Stadnik, Marciel J.;Mondino, Pedro
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.100-111
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    • 2019
  • Glomerella leaf spot (GLS) caused by Colletotrichum spp. is a destructive disease of apple restricted to a few regions worldwide. The distribution and evolution of GLS symptoms were observed for two years in Uruguay. The recurrent ascopore production on leaves and the widespread randomized distribution of symptoms throughout trees and orchard, suggest that ascospores play an important role in the disease dispersion. The ability of ascospores to produce typical GLS symptom was demonstrated by artificial inoculation. Colletotrichum strains causing GLS did not result in rot development, despite remaining alive in fruit lesions. Based on phylogenetic analysis of actin, ${\beta}$-tubulin and glyceraldehyde-3-phosphate dehydrogenase gene regions of 46 isolates, 25 from fruits and 21 from leaves, C. karstii was identified for the first time causing GLS in Uruguay and C. fructicola was found to be the most frequent (89%) and aggressive species. The higher aggressiveness of C. fructicola and its ability on to produce abundant fertile perithecia could help to explain the predominance of this species in the field.

Unreported Post-harvest Disease of Apples Caused by Plenodomus collinsoniae in Korea

  • Das, Kallol;Kim, Yeong-Hwan;Yoo, Jingi;Ten, Leonid N.;Kang, Sang-Jae;Kang, In-Kyu;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.511-518
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    • 2020
  • This study was conducted to isolate and identify the fungal pathogen caused unreported post-harvest disease on apples (cv. Fuji) fruit in Korea. The disease symptoms on apples appeared as irregular, light to dark brown, slightly sunken spots. The three fungal strains were isolated from infected tissues of apple fruits and their cultural and morphological characteristics were completely consistent with those of Plenodomus collinsoniae. The phylogenetic analysis using the internal transcribed spacer (ITS) regions, beta-tubulin (TUB), and the second largest subunit of RNA polymerase II (RPB2) sequences revealed the closest relationship of the isolates with Plenodomus collinsoniae at the species level. The pathogenicity test showed the same dark brown spots on Fuji apple cultivar. Therefore, P. collinsoniae is a newly reported fungal agent causing post-harvest disease on apples in Korea.

Isolation and Characterization of Eleven Unrecorded Pezizomycotina Species from Freshwater Ecosystems in Korea

  • Goh, Jaeduk;Jeon, Yu-Jeong;Mun, Hye Yeon;Chung, Namil;Park, Young-Hwan;Park, Sangkyu;Hwang, Hyejin;Cheon, Wonsu
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.423-443
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    • 2020
  • Freshwater fungi are a poly-phylogenetic group of taxonomically diverse organisms. In this study, we isolated diverse fungal strains from various environmental samples obtained from freshwaters in Korea. These strains were identified by performing molecular phylogenetic analyses of rDNA and/or other sequences (beta-tubulin, RNA polymerase II, and translation elongation factor 1). In addition, we examined their morphological characteristics microscopically and cultural characteristics using different media. We identified eleven unrecorded Pezizomycotina species: Cladosporium angulosum, Pseudorobillarda phragmitis, Paraconiothyrium estuarinum, Pseudopithomyces palmicola, Pyrenochaetopsis paucisetosa, Thelebolus globosus, Plagiostoma mejianum, Trichoderma cremeum, Fusarium tanahbumbuense, Coniochaeta endophytica, and Chaetomium tenue. Environmental samples obtained from different freshwater ecosystems in Korea could thus be a good source for isolating and investigating novel fungal species.

Diversity of the Bambusicolous Fungus Apiospora in Korea: Discovery of New Apiospora Species

  • Sun Lul Kwon;Minseo Cho;Young Min Lee;Hanbyul Lee;Changmu Kim;Gyu-Hyeok Kim;Jae-Jin Kim
    • Mycobiology
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    • v.50 no.5
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    • pp.302-316
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    • 2022
  • Many Apiospora species have been isolated from bamboo plants - to date, 34 bambusicolous Apiospora species have been recorded. They are known as saprophytes, endophytes, and plant pathogens. In this study, 242 bambusicolous Apiospora were isolated from various bamboo materials (branches, culms, leaves, roots, and shoots) and examined using DNA sequence similarity based on the internal transcribed spacer, 28S large subunit ribosomal RNA gene, translation elongation factor 1-alpha, and beta-tubulin regions. Nine Apiospora species (Ap. arundinis, Ap. camelliae-sinensis, Ap. hysterina, Ap. lageniformis sp. nov., Ap. paraphaeosperma, Ap. pseudohyphopodii sp. nov., Ap. rasikravindrae, Ap. saccharicola, and Ap. sargassi) were identified via molecular analysis. Moreover, the highest diversity of Apiospora was found in culms, and the most abundant species was Ap. arundinis. Among the nine Apiospora species, two (Ap. hysterina and Ap. paraphaeosperma) were unrecorded in Korea, and the other two species (Ap. lageniformis sp. nov. and Ap. pseudohyphopodii sp. nov.) were potentially novel species. Here, we describe the diversity of bambusicolous Apiospora species in bamboo organs, construct a multi-locus phylogenetic tree, and delineate morphological features of new bambusicolous Apiospora in Korea.

Molecular and Morphological Characterization of Three New Fungal Species of the Class Sordariomycetes from Korea

  • Kallol Das;Jae-Ho Ban;So-Young Choi;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.51 no.1
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    • pp.25-38
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    • 2023
  • Three fungal strains belonging to the class Sordariomycetes were isolated from soil collected on Jeju Island and Gyeongsangbuk-do, Korea. They were identified as Diaporthe endophytica (KNU-JJ-1809), Faurelina indica (KNU-JJ-1830), and Trichoderma ivoriense (KNU-4-KH1). KNU-JJ-1809 produced beta conidia that were straight, curved, hyaline, smooth-walled, with a diameter of 16.5-25.0×0.6-1.7 ㎛. The conidia of strain KNU-JJ-1830 were hyaline to light green, thin, clavate, round, truncate base, had guttules at both ends, with a diameter of 2.5-5.2×1.7-3.8 ㎛. The conidia of strain KNU-4-KH1 were oblong or ellipsoidal, smooth-walled, greenish, with a diameter of 2.2-4.4×2.2-3.6 ㎛. Internal transcribed spacer regions, partial large subunit, translation elongation factor 1-alpha, β-tubulin, and calmodulin genes were used to confirm the strains, and their cultural and morphological characteristics. To our knowledge, this is the first report on D. endophytica, F. indica, and T. ivoriense in Korea.

Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (Anser anser domestica) Gene Expression

  • Ji, Hong;Wang, Jianfa;Liu, Juxiong;Guo, Jingru;Wang, Zhongwei;Zhang, Xu;Guo, Li;Yang, Huanmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.3
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    • pp.423-432
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    • 2013
  • Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.

Selection and evaluation of reference genes for gene expression using quantitative real-time PCR in Mythimna separata walker (Lepidoptera: Noctuidae)

  • ZHANG, Bai-Zhong;LIU, Jun-Jie;CHEN, Xi-Ling;YUAN, Guo-Hui
    • Entomological Research
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    • v.48 no.5
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    • pp.390-399
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    • 2018
  • In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

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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