• 제목/요약/키워드: ITS rDNA

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First Report of Root Rot Caused by Plectosphaerella cucumerina on Cabbage in China

  • Li, Pan-Liang;Chai, A-Li;Shi, Yan-Xia;Xie, Xue-Wen;Li, Bao-Ju
    • Mycobiology
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    • 제45권2호
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    • pp.110-113
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    • 2017
  • Severe root rot was observed in fields of cabbages (Brassica oleracea L.) in 2015 in China. Cardinal symptoms of this disease included root rot and wilting leaves. A fungus was isolated from diseased tissues consistently. Based on the morphological features and molecular analysis of the ITS-5.8S rDNA and D1/D2 domain of the 28S rRNA gene, it was identified as Plectosphaerella cucumerina. This is the first report of P. cucumerina causing cabbage root rot in China and the world.

Phylogenetic Relationships of the Mutualistic Fungi Associated with Macrotermes subhyalinus in Oman

  • Hilal S. AlShamakhi;Abdullah M. Al-Sadi;Lyn G. Cook
    • Mycobiology
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    • 제51권5호
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    • pp.281-287
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    • 2023
  • The symbiotic association between fungus-gardening termites Macrotermes and its fungal symbiont has a moderate degree of specificity-although the symbiotic fungi (Termitomyces) form a monophyletic clade, there is not a one-to-one association between termite species and their fungus-garden associates. Here, we aim to determine the origin and phylogenetic relationships of Termitomyces in Oman. We used sequences of the internal transcribed spacer region (ITS) and the nuclear large subunit ribosomal RNA (LSU rRNA, 25S) gene and analyzed these with sequences of Termitomyces from other geographic areas. We find no evidence for more than a single colonization of Oman by Termitomyces. Unexpectedly, we find Termitomyces in Oman is most closely related to the symbiont of M. subhyalinus in West Africa rather than to those of geographically closer populations in East Africa.

LITHOAUTOTROPHIC NITROGEN REMOVAL WITH ANAEROBIC GRANULAR SLUDGE AS SEED BIOMASS AND ITS MICROBIAL COMMUNITY

  • Ahn, Young-Ho;Lee, Jin-Woo;Kim, Hee-Chul;Kwon, Soo-Youl
    • Environmental Engineering Research
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    • 제11권4호
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    • pp.173-180
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    • 2006
  • Autotrophic nitrogen removal and its microbial community from a laboratory scale upflow anaerobic sludge bed reactor were characterized with dynamic behavior of nitrogen removal and sequencing result of molecular technique (DNA extraction, PCR and amplification of 16S rDNA), respectively. In the experiment treating inorganic wastewater, the anaerobic granular sludge from a full-scale UASB reactor treating industrial wastewater was inoculated as seed biomass. The operating results revealed that an addition of hydroxylamine would result in lithoautotrophic ammonium oxidation to nitrite/nitrate, and also hydrazine would play an important role for the success of sustainable nitrogen removal process. Total N and ammonium removal of 48% and 92% was observed, corresponding to nitrogen conversion of 0.023 g N/L-d. The reddish brown-colored granular sludge with a diameter of $1{\sim}2\;mm$ was observed at the lower part of sludge bed. The microbial characterization suggests that an anoxic ammonium oxidizer and an anoxic denitrifying autotrophic nitrifier contribute mainly to the nitrogen removal in the reactor. The results revealed the feasibility on development of high performance lithoautotrophic nitrogen removal process with its microbial granulation.

Streptococcus pneumoniae가 생산하는 pneumolysin의 EGFP 융합으로 인한 용혈활성 변화 (C-terminal Fusion of EGFP to Pneumolysin from Streptococcus pneumoniae modified its Hemolytic Activity)

  • 정경태;이재헌;조혜주
    • 생명과학회지
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    • 제28권1호
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    • pp.99-104
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    • 2018
  • Streptococcus pneumoniae는 pneumolysin과 같은 병원성 인자를 가지고 있으며, 지역사회에서 전파되는 심각한 병원성균에 포함된다. Pneumolysin (PLY)은 콜레스테롤 의존적으로 세포막에 구멍을 형성하는 세포독소로서 백신의 주요한 표적 항원이다. PLY의 연구를 위하여 Streptococcus pneumoniae D39 균주에서 추출한 genomic DNA를 주형으로 하여 PCR을 시행하였다. 합성된 PLY 유전자 DNA를 pQE-30 vector에 삽입하고, E. coli M15에 형질전환 시킨 후 LB 배지에 IPTG를 첨가하여 PLY 단백질을 생산하였다. 재조합 단백질은 $Ni^{2+}$-agarose column을 사용하여 정제하였다. 또한, EGFP를 PLY C-말단에 부착한 융합단백질도 동일한 방법으로 클로닝하여 재조합 단백질을 생산하였다. 500 ng/ml 농도의 재조합 PLY는 1.0% 적혈구 현탁액을 100% 용혈시켰으며, 240 ng/ml 농도는 50% 용혈을 나타내었다. 그러나 재조합 PLY-EGFP는 용혈 활성이 전혀 나타나지 않았으나 형광현미경으로 관찰하였을 때 적혈구 막에 결합되어 있었다. 즉, EGFP의 PLY C-말단 부착은 PLY의 세포막 결합능은 유지시켰으나 용혈기능은 방해하였다. PLY C-말단은 용혈기능에 아주 중요한 영역이며, 세포막 결합은 PLY의 다른 영역이 보다 중요하게 작용할 것으로 추측된다. 따라서, 육안으로 관찰이 가능한 결합능은 가졌으나 용혈 기능이 결여된 PLY-EGFP를 대조군으로 활용함으로써 두 재조합 단백질은 폐렴 유발에 있어서 PLY 작용 연구에 사용될 수 있을 것으로 기대된다.

Sclerotium rolfsii에 의한 녹두 흰비단병 (Sclerotium Rot of Mungbean (Phaseolus radiatus L.) Caused by Sclerotium rolfsii in South Korea)

  • 권진혁;김민근;강동완;한인영;이병정;김진우
    • 한국균학회지
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    • 제45권3호
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    • pp.246-250
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    • 2017
  • 2015년 9월 경상남도농업기술원 시험연구포장에 재배중인 녹두에서 흰비단병 증상이 발생하였다. 병징은 녹두의 줄기 지제 부위가 수침상으로 물러지고 부패되어 서서히 시들면서 포기 전체가 말라 죽었다. 줄기의 병반부와 토양 표면에 흰색의 곰팡이가 발생하며 갈색의 작은 둥근 균핵을 많이 형성하였다. 감자한천배지에서 균총은 흰색이고 잘 자라며 배양기간이 경과됨에 따라 갈색의 작은 둥근 균핵을 많이 형성하였다. 균핵의 크기는 1~3 mm이며 균사의 폭은 $4{\sim}9{\mu}m$였다. 균사 생육과 균핵 형성 적온은 $30^{\circ}C$이었다. 균사 특유의 clamp connection이 관찰되었다. 녹두에서 발생한 병징과 병원균의 균학적 특징, internal transcribed spacer rDNA 염기서열 비교분석 결과를 토대로 이 병을 Sclerotium rolfsii Saccardo에 의한 녹두 흰비단병으로 명명하고자 제안한다.

Identification of Clostridium perfringens AB&J and Its Uptake of Bromophenol Blue

  • Kim, Jeong-Dong;An, Hwa-Yong;Yoon, Jung-Hoon;Park, Yong-Ha;Fusako Kawai;Jung, Chang-Min;Kang, Kook_-Hee
    • Journal of Microbiology and Biotechnology
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    • 제12권4호
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    • pp.544-552
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    • 2002
  • Several microorganisms from rat and human feces and lumen fluid of cows were screened for their ability to decolorize the synthetic dyes. Consequently, a novel dye-degrading strain AB&J was isolated. Taxonomic identification including 165 rDNA sequencing and phylogenetic analysis indicated that the isolate had 99.9% homology in its 165 rDNA base sequence with Clostridium perfringens. After 27 h Incubation with the strain, brilliant blue R, bromophenol blue, crystal violet, malachite green, methyl green, and methyl orange were decolorized by about 69.3%, 97.7%, 96.3%, 97.9%, 75.1%, and 97.2%, respectively. The triphenlmethane dye, bromophenol blue, was decolorized extensively by growing Clostridium perfringens AB&J cells in liquid cultures under anaerobic condition, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly decolorized at a relatively lower concentration of below 50 $\mu g \;ml^{-1}$, however, the growth of the cells was mostly suppressed at a dye concentration of 100 $\mu g \;ml^{-1}$. The decolorization activity in cell-free extracts was much higher in cytoplasm than in periplasm and cytoplasmic membrane. Therefore, the enzyme related uptake of bromophenol blue seemed to be localized in cytoplasm. The optimal pH and temperature of bromophenol blue uptake fur decolorization activities were 7.0 and 4$0^{\circ}C$, respectively.

Identification and Characterization of an Antifungal Protein, AfAFPR9, Produced by Marine-Derived Aspergillus fumigatus R9

  • Rao, Qi;Guo, Wenbin;Chen, Xinhua
    • Journal of Microbiology and Biotechnology
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    • 제25권5호
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    • pp.620-628
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    • 2015
  • A fungal strain, R9, was isolated from the South Atlantic sediment sample and identified as Aspergillus fumigatus. An antifungal protein, AfAFPR9, was purified from the culture supernatant of Aspergillus fumigatus R9. AfAFPR9 was identified to be restrictocin, which is a member of the ribosome-inactivating proteins (RIPs), by MALDI-TOF-TOF-MS. AfAFPR9 displayed antifungal activity against plant pathogenic Fusarium oxysporum, Alternaria longipes, Colletotrichum gloeosporioides, Paecilomyces variotii, and Trichoderma viride at minimum inhibitory concentrations of 0.6, 0.6, 1.2, 1.2, and 2.4 μg/disc, respectively. Moreover, AfAFPR9 exhibited a certain extent of thermostability, and metal ion and denaturant tolerance. The iodoacetamide assay showed that the disulfide bridge in AfAFPR9 was indispensable for its antifungal action. The cDNA encoding for AfAFPR9 was cloned from A. fumigatus R9 by RT-PCR and heterologously expressed in E. coli. The recombinant AfAFPR9 protein exhibited obvious antifungal activity against C. gloeosporioides, T. viride, and A. longipes. These results reveal the antifungal properties of a RIP member (AfAFPR9) from marine-derived Aspergillus fumigatus and indicated its potential application in controlling plant pathogenic fungi.

Evaluation of MiR-34 Family and DNA Methyltransferases 1, 3A, 3B Gene Expression Levels in Hepatocellular Carcinoma Following Treatment with Dendrosomal Nanocurcumin

  • Chamani, Fatemeh;Sadeghizadeh, Majid;Masoumi, Mahbobeh;Babashah, Sadegh
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권sup3호
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    • pp.219-224
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    • 2016
  • Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver making up more than 80 percent of cases. It is known to be the sixth most prevalent cancer and the third most frequent cause of cancer related death worldwide. Epigenetic regulation constitutes an important mechanism by which dietary components can selectively activate or inactivate target gene expression. The miR-34 family members including mir-34a, mir-34b and mir-34c are tumor suppressor micro RNAs, which are expressed in the majority of normal tissues. Several studies have indicated silencing of miR-34 expression via DNA methylation in multiple types of cancers. Bioactive nutrients like curcumin (Cur) have excellent anticarcinogenic activity and minimal toxic manifestations in biological systems. This compound has recently been determined to induce epigenetic changes. However, Cur is lipophilic and has a poor systemic bioavailability and poor absorption. Its bioavailability is increased through employing dendrosome nanoparticles. The aim of the current study was to investigate the effect of dendrosomal nanocurcumin (DNC) on expression of mir-34 family members in two HCC cell lines, HepG2 and Huh7. We performed the MTT assay to evaluate DNC and dendrosome effects on cell viability. The ability of DNC to alter expression of the mir-34 family and DNA methyltransferases (DNMT1, DNMT3A and 3B) was evaluated using semi-quantitative and quantitative PCR. We observed the entrance of DNC into HepG2 and Huh7 cells. Gene expression assays indicated that DNC treatment upregulated mir34a, mir34b and mir34c expression (P<0.05) as well as downregulated DNMT1, DNMT3A and DNMT3B expression (P<0.05) in both HepG2 and Huh7 cell lines. DNC also reduced viability of Huh7 and HepG2 cells through restoration of miR-34s expression. We showed that DNC could awaken the epigenetically silenced miR-34 family by downregulation of DNMTs. Our findings suggest that DNC has potential in epigenetic therapy of HCC.

옥수수 종피의 안토시아닌 합성을 조절하는 R 유전자 구성요소의 구명 (Identification of the Maize R Gene Component Responsible for the Anthocyanin Biosynthesis of Kernel Pericarp)

  • 김화영
    • 한국육종학회지
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    • 제42권1호
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    • pp.50-55
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    • 2010
  • 옥수수 R 유전자의 대립인자 중 하나인 R-r:standard (R-r:std)는 종자 호분층의 안토시아닌 합성을 조절하는 S subcomplex와 종자 이외 식물체부위의 안토시아닌 합성을 조절하는 P component로 구성되어 있으며, S subcomplex는 S1 및 S2 component로 구성되어 있다. R 유전자의 대립인자 중 일부는 Pl 유전자가 존재할 경우 종피의 안토시아닌 합성을 유도한다. 따라서 Pl 유전자가 존재할 경우 옥수수 종피의 안토시아닌 합성을 유도하는 R 유전자의 구성요소를 구명하고자 종피의 안토시아닌 합성에 미치는 서로 다른 R 인자들의 효과를 분석하였다. R-ch와 r-ch 인자는 유사한 정도의 착색 효과를 보였으며, R-r:Ecuador (R-r:Ec)는 이들보다 짙은 착색효과를 나타내었다. S subcomplex의 기능은 상실하였으나 정상적인 P component를 보유하고 있는 것으로 추정되는 r-ch는 Pl 유전자가 존재할 경우 종피의 색소합성 기능을 유지하고 있으나, S subcomplex의 기능은 정상이나 P component의 기능은 잃어버린 것으로 추정되는 R-r:Ec 유래 인자 R-g:g1111는 Pl 유전자가 존재할 경우에도 종피가 착색되지 않았다. 더욱이 R-ch와 r-ch 인자의 PCR 분석 결과, R-ch는 P와 S1 component를 보유하고 있으나, r-ch는 S1을 보유하고 있지 않는 것으로 나타났다. 따라서 R 유전자의 구성요소 중 P component가 종피의 안토시아닌 합성에 관여하는 구성요소로 추정되었다.

Microstroma juglandis에 의한 호두나무 흰곰팡이병 발생 (Occurrence of Downy Leaf Spot on Juglans regia Caused by Microstroma juglandis in Korea)

  • 이상현;이승규;박지현;조성은;신현동
    • 식물병연구
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    • 제17권3호
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    • pp.386-390
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    • 2011
  • 2011년 6월에 김천과 무안의 호두나무 재배지에서 잎에 노균병 병징과 유사한 증상이 관찰되었다. 병든 잎의 뒷면에는 다각형의 병반에 흰색의 곰팡이가 눈꽃처럼 형성되었고, 상응하는 잎의 앞면에는 퇴록 부분이 생겼다. 병세가 진전되면서 병든 조직은 붕괴되었고 나중에는 괴사되었다. 이 병의 원인균은 형태적 및 배양적 특징으로 보아 Microstroma juglandis로 동정되었다. 또한 균주의 ITS rDNA 염기서열을 GenBank에 등록된 M. juglandis의 자료와 비교하여 100% 일치함을 확인하였다. 호두나무 묘목에 병원성 검정을 실시하여 코흐의 원칙을 만족시켰다. 이 병은 지금까지 북미, 유럽, 오세아니아 그리고 서아시아(인도, 파키스탄, 터키)에서 보고되었다. 이 연구를 통하여 한국을 포함한 동아시아에서는 이 병을 처음으로 보고하며, 병명은 '호두나무 흰곰팡이병'으로 제안한다.