• Title/Summary/Keyword: 분자 계통분석

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버섯의 항암효과 -영자 균사체 다당류의 항종양 활성-

  • 이준우
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.13-14
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    • 1997
  • 영지의 약리효과는 triterpenoid계의 저분자와 polysaccharide인 고분자로 대별할 수 있다. 저분자 물질은 항염증작용, 항알러지작용, 간암세포 성장 억제작용, 혈압 강하작용, 혈소판 응집저해작용 등이 있으며, 고분자 물질은 혈압강하작용, 정혈작용, 고지혈증 개선작용, 혈당 강하작용, 면역 증강작용 및 항종양작용 등이 있는 것으로 알려졌다. 현재까지의 영지에 관한 연구는 주로 자실체에 대한 연구가 주류를 이루고 있으며 이들의 산지, 환경조건, 채집시기 및 실험자들에 따라 약리 효과 등에 많은 차이가 있다. 이들에 대한 효과가 영지의 대표적 약리효과라고 언급하기엔 부족한 점이 많으므로 약리효과가 우수하고 일관성이 있는 영지 유래 다당류를 얻기 위한 방법으로서는 균사체의 대량배양법 및 분리방법이 확립 되어야만 한다. 본 연구에서는 새로운 항암활성 다당류의 개발을 위해 국내 자생 영지를 채집하여 검색 및 계통적 분류를 행하였으며 분리된 균주의 최적 액체배양 조건, 다당류 분리 조건, 다당류의 물리화학적 특성 및 약리활성 등에 관한 연구를 실시하였다. 그 중 약리활성이 우수한 G- lucidum IY009 균주를 선발 하였다. 구조적 특징과 종양 면역 활성과의 관계 규명을 위해, G- lucidum IY009 배양균사체로부터 다당류를 T-AS, U-AS, M-AS, S 및 H로 분획 하였다. 추출조건에 따른 다당류 GLG의 수율은 열수 추출 분획이 2.98g/l로 가장 높았으며, U-AS및 T-AS에서 각각 2.32g/l, 2.07g/l이었다. GLG의 특성을 조사하기 위하여 수용성과 비수용성으로 분리한 결과 수용성 분획은 비수용성 분획보다 높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정 혈청단백을 증가시켰으며, 항체 생성능의 증가가 관찰되어 effect T 세포의 활성화가 나타나고 있음을 알 수 있었다. T-AS는 생체내 투여시에 대식세포의 탐식능이 증진되었으며, 대식세포 기능 저해제에 의한 대식세포의 기능 저해 현상이 회복되었다. 이와 같은 결과들로부터, T-AS의 항암 활성은 활성화된 보체 성분 및 당 수용체들이 존재하는 대식세포의 개입을 시사한다.

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Study on the Genetic Characteristics of Waterlogging Tolerant Pepper (Capsicum annuum L.) for Breeding Tolerant Varieties against Flooding Stress (내습성 고추 품종 육성을 위한 선발계통의 유전적 특성 구명)

  • Yang, Eun Young;Chae, Soo-Young;Hong, Jong-Pil;Lee, Hye-Eun;Park, Eun Joon;Moon, Ji-hye;Park, Tae-Sung;Roh, Mi-Young;Kim, Ok Rye;Kim, Sang Gyu;Kim, Dae Young;Lee, Sun Yi;Cho, Myeong Cheoul
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1111-1120
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    • 2017
  • This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

Two Unrecorded Endophytic Fungi Isolated from Funaria hygrometrica in Korea (표주박이끼(Funaria hygrometrica)에서 분리된 2종의 국내 미기록 내생균)

  • Choi, Hyun-Sook;Park, Hyeok;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.313-318
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    • 2019
  • In the present study, we isolated endophytic fungal strains from the rhizoids of the moss Funaria hygrometrica. The isolated strains were identified based on morphological characteristics and analysis of the internal transcribed spacer (ITS) and large subunit (LSU) rDNA sequence regions. Consequently, we confirmed the presence of two endophytic fungal species, Curvularia protuberata and Didymella anserina, which have not been reported in Korea previously. Here, we describe the morphological characteristics and molecular analysis results of these fungal species.

Unrecorded Endophytic Fungi Isolated from Mnium heterophyllum and Hypnum plumaeforme in Korea: Biscogniauxia petrensis and Cercophora thailandica (꼬마초롱이끼(Mnium heterophyllum)와 가는털깃털이끼(Hypnum plumaeforme)에서 분리한 국내 미기록 내생균: Biscogniauxia petrensis, Cercophora thailandica)

  • Choi, Hyun-sook;Park, Hyeok;Eo, Ju-Kyeong;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.48 no.1
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    • pp.55-61
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    • 2020
  • In this study, we isolated endophytic fungal strains from the rhizoid and the leaf of Mnium heterophyllum and Hypnum plumaeforme, respectively. The isolated strains were identified based on morphological characters and molecular analysis of the internal transcribed spacer, large subunit rDNA, β-tubulin, and RNA polymerase II subunit regions. From the results, we confirmed two endophytic fungal species, Cercophora thailandica, and Biscogniauxia petrensis, which to the best of our knowledge, have not yet been reported in Korea. We further describe the morphological characteristics and the results of the molecular analyses of these isolated fungal strains.

Pulmonary Candidiasis in a Dog Receiving Long-term Management for Atopic Dermatitis (아토피성 피부염의 장기 치료중인 개에서 발생한 폐 칸디다증)

  • Han, Jae-Ik;Lee, Chang-Min;Kim, Taewoo;Lee, In;Choi, Soo-Young;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.32 no.3
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    • pp.259-262
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    • 2015
  • A canine patient exhibited partial anorexia and sudden respiratory distress. Diagnostic imaging and cytology of tracheal-lavage fluids revealed fungal pneumonia. Molecular identification and phylogenetic analysis detected Candida catenulata. Treatment with oral itraconazole for 3 weeks was effective. This is the first report of C. catenulata infection in a dog.

Three Novel Endophytic Fungal Species Isolated from Roots of Medicinal Crops in Korea (약용작물의 뿌리에서 분리된 3종의 국내 미기록 내생균)

  • Park, Hyeok;Jung, Chung Ryul;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.47 no.2
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    • pp.113-120
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    • 2019
  • We isolated endophytic fungal strains from the roots of three medicinal crops, Ligusticum chuanxiong, Angelica gigas, and Cnidium officinale, cultivated at Yeongju, Korea. The fungal strains were identified based on their morphological characteristics and molecular analyses of their internal transcribed spacer, large subunit rDNA, and beta-tubulin regions. Thereby, we identified three previously unrecorded endophytic fungal species, Dactylonectria pauciseptata, Rhizopycnis vagum, and Sistotrema sernanderi, in Korea. In this report, we describe the morphological characteristics and phylogenetic analyses of these three fungal species.

Unrecorded mycoparasitic fungus Sepedonium laevigatum isolated from wild mushrooms in South Korea (야생버섯에서 분리한 미기록종 버섯기생균 Sepedonium laevigatum 보고)

  • Seung-Yoon, Oh
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.249-253
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    • 2022
  • Fungi that infect mushrooms and cause diseases are called mycoparasitic fungi. Understanding the diversity of mycoparasitic fungi in wild mushrooms is important for the mushroom industry, as they can parasitize cultivated mushrooms. However, few studies have been conducted on the diversity of mycoparasitic fungi in wild mushrooms in South Korea. Upon surveying mushroom flora, we found a mycoparasitic fungus and analyzed its strain using molecular phylogeny and morphological examination. The strain was identified as Sepedonium laevigatum, which is an unrecorded species in South Korea. In this study, we described the cultural and microscopic characteristics of this strain of fungus.

Phylogeny of the Yeast Species Isolated from Wild Tiger Lily (Lilium lancifolium Thunb.) (야생 참나리(Lilium lancifolium Thunb.)로부터 분리한 효모의 분자계통학적 분석)

  • Kim, Jong-Shik;Kim, Dae-Shin
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.149-154
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    • 2015
  • BACKGROUND: Yeast isolates associated with the leaves, stems, and flowers of the tiger lily needed to be identified using isolation methods that have previously been used effectively in yeast biotechnology. A culture-based approach was necessary for the isolation of many yeast strains associated with tiger lily. METHODS AND RESULTS: In this study, the homogenized leaves, stems, and flowers of tiger lily were spreaded onto GPY medium containing chloramphenicol, streptomycin, Triton X-100, and L-sorbose. A total of 82 yeast strains from the leaves, 94 and 97 yeast strains from the stems and flowers were isolated, respectively. Yeast isolates were identified by phylogenetic analysis based on internal transcribed spacer region sequencing. The yeast species isolated from the leaves comprised of 31 isolates of the genus Pseudozyma, 28 of Aureobasidium pullulans, and 11 of the genus Cryptococcus. Those isolated from the stems comprised of 40 of A. pullulans and 11 of Cryptococcus, and 95 of A. pullulans While, 1 isolate each of the genera Rhodotorula and Metschnikowia were isolated from the flowers. CONCLUSION: We identified site-specific yeast communities associated with tiger lily. These yeast isolates may have high potential for application in the field of biotechnology.

Incidence of Aphid-Transmitted Strawberry Viruses in Korea and Phylogenetic Analysis of Korean Isolates of Strawberry Mottle Virus (진딧물 전반 딸기 바이러스 발생조사 및 딸기모틀바이러스의 계통분석)

  • Kwon, Sun-Jung;Yoon, Jung-Beom;Cho, In-Sook;Yoon, Ju-Yeon;Kwon, Tae-Ryong
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.226-232
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    • 2019
  • Among more than 30 viruses infecting strawberry, aphid-transmitted viruses including Strawberry mild yellow edge virus (SMYEV), Strawberry mottle virus (SMoV), Strawberry crinkle virus (SCV), and Strawberry vein banding virus (SVBV) have been considered as the most economically important viruses of strawberry in the world. To determine the incidence of these four viruses in major Korean strawberry cultivars, field surveys were conducted in major production areas during 2018-2019. In our surveys, SMYEV and SMoV were detected with low infection rates of 0.7% and 1.3%, respectively, whereas SCV and SVBV were not detected. No obvious symptoms were observed in the strawberry plants infected by SMYEV or SMoV. Since no sequences of SMoV Korean isolates have been reported, we initially determined nucleotide sequence of the 3' untranslated region (UTR) of seven SMoV isolates obtained during the surveys. The 3' UTR sequences (782 nt) of seven Korean isolates were phylogenetically compared with those of the previously reported SMoV isolates. Phylogenetic analysis revealed that most Korean isolates are closely related to Canadian isolates and only little evolutionary differentiation occurred among the Koreans isolates. This might be due to the low incidence of SMoV in strawberry in Korea.

Isolation and Identification of Halotolerant Bacillus sp. SJ-10 and Characterization of Its Extracellular Protease (세포외 Protease를 생산하는 내염성 Bacillus sp. SJ-10 균주의 분리 동정 및 효소 특성)

  • Kim, Eun-Young;Kim, Dong-Gyun;Kim, Yu-Ri;Choi, Sun-Young;Kong, In-Soo
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.193-199
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    • 2009
  • A bacterium producing the halotolerant extracellular protease was isolated from squid jeotgal, and was identified as Bacillus sp. SJ-10 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The strain grew at $20^{\circ}C\sim55^{\circ}C$, pH 5~8, and 0%~14% NaCl and optimal growth conditions were $35{\pm}5^{\circ}C$, pH 7, and 5% NaCl. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and $C_{16:0}$ DNA G+C content was 50.58 mol% and menaquinone consisted of MK-7 Phylogenic analysis based on the 16S rRNA gene sequence indicated that SJ-10T belongs to the genus Bacillus. About 40 kDa of the salt-tolerant protease was purified by 40% ammonium sulfate saturation and Mono Q column chromatography. The optimal activity of the protease was pH 8 and stable at pH 5~10. The optimum temperature and NaCl concentration were $35{\pm}5^{\circ}C$ and $5{\pm}1%$, respectively.