• 제목/요약/키워드: chitin synthase gene

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Characterization and Phylogenetic Analysis of Chitin Synthase Genes from the Genera Sporobolomyces and Bensingtonia subrorea

  • Nam, Jin-Sik
    • 환경생물
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    • 제23권4호
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    • pp.335-342
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    • 2005
  • We cloned seven genes encoding chitin synthases (CHSs) by PCR amplification from genomic DNAs of four strains of the genus Sporobolomyces and of Bensingtonia subrosea using degenerated primers based on conserved regions of the CHS genes. Though amino acid sequences of these genes were shown similar as 176 to 189 amino acids except SgCHS2, DNA sequences were different in size, which was due to various introns present in seven fragments. Alignment and phylogenetic analysis of their deduced amino acid sequences together with the reported CHS genes of basidiomycetes separated the sequences into classes I, II and III. This analysis also permitted the classification of isolated CHSs; SgCHS1 belongs to class I, BsCHS1, SaCHS1, SgCHS2, SpgCHS1, and SsCHS1 belong to class II, and BsCHS2 belongs to class III. The deduced amino acid sequences involving in class II that were discovered from five strains were also compared with those of other basidiomycetes by CLUSTAL X program. The bootstrap analysis and phylogenetic tree by neighbor-joining method revealed the taxonomic and evolutionary position for four strains of the genus Sporobolomyces and for Bensingtonia subrosea which agreed with the previous classification. The results clearly showed that CHS fragments could be used as a valuable key for the molecular taxonomic and phylogenetic studies of basidiomycetes.

무 탄저병을 일으키는 Colletotrichum속의 종 동정 및 병원성 (Identification and Pathogenicity of Colletotrichum Species Causing Anthracnose of Korean Radish (Rhaphanus sativus) in Korea)

  • 최효원;김점순;홍성기;이영기
    • 한국균학회지
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    • 제47권4호
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    • pp.393-406
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    • 2019
  • 2017년 8월경, 강원도 홍천군, 정선군, 평창군 소재 무 재배 포장에서 잎자루와 잎의 주맥에 형성된 탄저병 병징이 관찰되었다. 회색 내지 짙은 갈색을 띤 1-2 mm 크기의 점무늬가 아래 잎과 잎의 주맥에 형성되었다. 이러한 점무늬는 점차 확대되고 합쳐지면서 갈색을 띤 불규칙한 병반으로 나타났다. 병든 무 잎에서 10개의 Colletotrichum균을 분리하였다. 분리 균주의 형태적 특성과 internal transcribed spacers and intervening 5.8S rDNA (ITS), partial beta-tubulin gene (TUB2), partial actin gene (ACT) 및 partial chitin synthase-1 gene (CHS-1) 부위를 사용한 다자위 분자 계통 분석(multilocus molecular phylogenetic analysis)에 의해 8개 균주는 Colletotrichum higginsianum, 2개 균주는C. truncatum으로 동정되었다. 무(시래기무, 오사리무)와 배추(품종: 추노, 스마트)를 대상으로 상처와 무상처로 구분하여 포자현탁액을 떨어뜨려서 접종하는 방법을 사용하여 병원성 검정을 수행하였다. 무에서는C. truncatum 1개 균주를 제외하고 상처와 무상처 접종에서 모두 병징을 나타냈고, 배추의 경우는C. higginsianum 접종구에서만 상처의 유무에 관계없이 병이 발생하였다. 국내에는 기존에 C. higginsianum은 무와 배추의 탄저병균으로 보고되었으며, C. truncatum에 의한 무 탄저병의 발생은 본 연구에서 국내 최초로 보고한다.

First Report of Anthracnose of Shine Muscat Caused by Colletotrichum fructicola in Korea

  • Lim, Yang-Sook;Hassan, Oliul;Chang, Taehyun
    • Mycobiology
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    • 제48권1호
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    • pp.75-79
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    • 2020
  • Anthracnose is one of the major problems for cultivating many crops, including vegetables, fruits, and trees. It is a continual threat for fruits grower worldwide. Colletotrichum fructicola was isolated from Shine Muscat berries showing typical anthracnose symptom in Korea. It was identified as C. fructicola based on morphology, pathological signs and concatenated sequences of internal transcribed spacer region of rDNA, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin-2, chitin synthase-1, calmodulin, and the Apn2-Mat1-2 intergenic spacer and partial mating type (Mat1-2) gene. To the best of our knowledge, this is the first report first report of anthracnose of Shine Muscat caused by C. fructicola in Korea.

Colletotrichum cymbidiicola Causing Anthracnose on Cymbidium Orchids in Korea

  • Park, Mi-Jeong;Back, Chang-Gi;Park, Jong-Han
    • Mycobiology
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    • 제48권4호
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    • pp.321-325
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    • 2020
  • A Colletotrichum species was isolated from leaves of Cymbidium exhibiting symptoms of anthracnose. In this study, the isolates obtained were identified based on recent taxonomic approaches for the genus Colletotrichum. The identity of the causal pathogen was confirmed using morphological data and phylogenetic analysis of combined multi-gene dataset (internal transcribed spacer, glyceraldehyde 3-phosphate dehydrogenase, chitin synthase-1, actin, histone3, beta-tubulin, and calmodulin). Pathogenicity testing revealed that the isolates were pathogenic to Cymbidium. Based on these results, the fungal pathogen occurring on Cymbidium orchids was identified as Colletotrichum cymbidiicola, which is a newly recorded species in Korea.

Colletotrichum fioriniae에 의한 호박 과실 탄저병의 발생 보고 (First Report of Colletotrichum fioriniae Causing Anthracnose on Fruit of Pumpkin (Cucurbita moschata) in Korea)

  • 김준영;김병섭
    • 식물병연구
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    • 제26권3호
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    • pp.190-193
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    • 2020
  • 강원도 강릉시 사천면 비닐하우스에서 재배중인 호박 과실에 탄저병이 발생하였다. 병든 호박 과실에 분홍색의 분생포자 층이 동심원으로 나타나 점차 확대되어 과실이 무르는 증상을 나타내었다. 원인균을 규명하기 위하여 순수 분리 후 균학적 특성 및 ITS, GAPDH, CHS-1, HIS3, ACT, TUB2 염기서열 분석결과 Colletotrichum fioriniae로 동정하였다. 또한, 병원성이 확인되었고 접종시험에서 동일한 균이 반복적으로 분리되었다. 따라서, 이러한 결과를 바탕으로 C. fioriniae에 의한 호박 과실에 발생하는 탄저병의 발생을 국내 처음으로 보고한다.

고온에서 배추좀나방 유충 지방체의 유전자 발현 변화 분석 (Analysis of Gene Expression in Larval Fat Body of Plutella Xylostella Under High Temperature)

  • 김광호;이대원
    • 한국환경농학회지
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    • 제37권4호
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    • pp.324-332
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    • 2018
  • 곤충은 변온동물로 육지생태계에서 주로 서식하면서, 식물의 생체량 조절, 종다양성 유지에 중요한 역할을 한다. 주변온도는 변온동물인 곤충의 생리적 반응속도, 뿐만 아니라 생존과 분포를 결정하며, 기후변화에 영향을 준다. 본 연구는 높은 온도에서 곤충의 적응성에 관련있는 유전자를 전사체를 이용하여 동정하였다. 고온에서 사육된 배추좀나방 유충의 지방체로부터 차세대 염기서열 분석법을 이용하여 전사체를 확보하였다. 대사중심인 지방체에서 구조단백질, 열충격단백질, 항산화단백질, 해독효소 들이 동정되었다. 이들 중에서 표피단백질(표피단백질, 키틴합성효소, 엑틴, 카이틴 합성), 스트레스관련단백질(시토크롬 P450), 열충격단백질, 한산화단백질은 발현이 증가되었으나, glutathione S transferase 발현은 오히려 감소되었다. 이상의 결과는 기후변화의 주요인인 온난화에 대한 해충의 생리적 대응과 온도적응을 이해하는데 필요한 기초자료를 제시한다.

가래나무 수액의 당성분 분석 및 생리활성 연구 (Studies on the Sugar Analysis and Biological Activity of Sap from Juglans mandshurica Maxim.)

  • 김대욱;최미나;이민성;정회석;변준기;김영수
    • 한국키틴키토산학회지
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    • 제23권4호
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    • pp.262-266
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    • 2018
  • 우리나라토종호두나무인가래나무(Juglans mandshurica Maxim.)의 수액의 유리당 함량은 약 3%로 다른 나무 수액에 비해 높게 나타났으며 미량 원소 중에는 Ca이 30 mg/kg의 농도로 포함되어 있는 것으로 확인되었다. 가래나무 수액 $200{\mu}g/ml$에서 elastase 저해 활성이 약 30% 정도를 보이고 있고 HaCaT 세포를 이용한 피부보습 효과에서는 자외선을 조사한 HaCaT세포의 aquaporin 3 유전자와 hyaluronan synthase 2 유전자 발현을 촉진시켜 hyaluronic acid의 생합성을 증가시킨 것으로 확인되었다. 본 연구를 통해 가래나무 수액의 피부 노화 방지 효능을 최초로 규명하였고 이는 가래나무 수액의 활용도를 높일 수 있는 가능성을 제시한 결과라 사료된다. 다만 물질 분석과 in-vivo 시험, 임상 연구 등 심도 있는 추가 연구가 필요할 것으로 판단된다.

Colletotrichum fructicola, a Member of Colletotrichum gloeosporioides sensu lato, is the Causal Agent of Anthracnose and Soft Rot in Avocado Fruits cv. "Hass"

  • Fuentes-Aragon, Dionicio;Juarez-Vazquez, Sandra Berenice;Vargas-Hernandez, Mateo;Silva-Rojas, Hilda Victoria
    • Mycobiology
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    • 제46권2호
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    • pp.92-100
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    • 2018
  • The filamentous Ascomycota Colletotrichum gloeosporioides sensu lato is a fungus that has been reported worldwide as a causal agent of anthracnose disease in avocado and other crops. In Mexico, this species affects fruits from an early stage of development in the orchard until the post-harvest stage. Although fungicides are continuously applied to control Colletotrichum species, pericarp cankers and soft rot mesocarp in fruits are still frequently observed. Considering the lack of a precise description of the causative agent, the aim of the current study was to determine the pathogens involved in this symptomatology. Twenty-four isolates were consistently obtained from the pericarp of avocado fruits cv. "Hass" collected in the central avocado-producing area of Mexico. Morphological features such as colony growth, conidia size, and mycelial appressorium were assessed. Bayesian multilocus phylogenetic analyses were performed using amplified sequences of the internal transcribed spacer region of the nuclear ribosomal DNA; actin, chitin synthase, glyceraldehyde-3-phosphate dehydrogenase partial genes; and APn2-Mat1-2 intergenic spacer and mating type Mat1-2 partial gene from the nine selected isolates. In addition, fruits were inoculated with a conidial suspension and reproducible symptoms confirmed the presence of Colletotrichum fructicola in this area. This pathogenic species can now be added to those previously reported in the country, such as C. acutatum, C. boninense, C. godetiae, C. gloeosporioides, and C. karstii. Disease management programs to reduce the incidence of anthracnose should include C. fructicola to determine its response to fungicides that are routinely applied, considering that the appearance of new species is affecting the commercial quality of the fruits and shifting the original population structure.

온도변화에 따른 담배나방 유충 지방체의 유전자 발현 비교 분석 (Comparison of Gene Expression in Larval Fat Body of Helicoverpa assulta in Different Temperature Conditions)

  • 차욱현;김광호;이대원
    • 한국응용곤충학회지
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    • 제57권3호
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    • pp.165-175
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    • 2018
  • 곤충은 넓은 범위의 온도영역에 사는 것으로 알려져 있으나, $40^{\circ}C$가 넘는 고온이나 빙결온도 이하의 저온에서는 생존이 어렵다. 본 연구는 사육온도 조건이 다른 환경에서 대사중심 조직인 지방체의 유전자 발현을 분석하기 위해, 온도조건을 달리하여 담배나방을 저온 사육충 ($3{\sim}10^{\circ}C$), 고온 사육충 ($35^{\circ}C$)로 나누고 상온 사육충 ($25^{\circ}C$)을 대조구로 사용하여 전사체 분석을 수행하였다. 저온에서 특이적으로 높은 발현을 보인 유전자는 표피단백질, ${\Delta}9$ 불포화효소, 글리세롤 3-인산 탈수소효소이며, 저온에서 발현이 낮아진 유전자는 키틴 합성효소, catalase, UDP-당전이 효소이다. 고온에서 특이적으로 높은 발현을 보인 유전자는 과산화물제거효소, metallothionein 2, phosphenolpyruvate carboxykinase, 트레할로스 운반단백질이었다. 고온에서 높고 저온에서 낮은 대조적 발현을 보인 유전자는 열충격단백질, glutathione peroxidase이었다. 이들 온도 특이적이거나 대조적 발현을 보이는 유전자는 기후변화에 관련한 특이마커로 활용이 가능할 것으로 사료된다.

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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    • 한국균학회 2014년도 추계학술대회 및 정기총회
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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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