一株南海指海绵共附生真菌多样性及其抗菌活性
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摘要
本研究通过采用6种不同的培养基对南海指海绵上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究海绵共附生真菌多样性。共分离培养出55株(21种)真菌菌株,优势菌属为木霉菌属Trichoderma sp.(31株),其中的哈茨木霉Trichoderma harzianum 22株作为优势菌种,占真菌菌株总数的40%。其余真菌则属于枝孢菌属Cladosporium、迈耶氏酵母属Meyerozyma、茎点霉属Phoma、红酵母属Rhodotorula、青霉菌属Penicillium、曲霉属Aspergillus等10个属,还有两株未鉴定到种的子囊菌。选取金黄色葡萄球菌Staphylococcus aureus等7种病原指示菌,采用琼脂扩散法对21种真菌的代表性菌株进行抗菌活性筛选,其中有3种真菌至少对一种指示菌表现出抑菌效果,占菌株种类总数的11%。
Six different kinds of culture media are used to isolate and cultivate the fungi associated with the South China Sea sponge. The isolates obtained totaled 55, belonging to 22 species. The identification wasmade through morphological observation in combination with ITS-rDNA blast homology analysis. The dominant genus is Trichoderma(31 isolates), and Trichoderma harzianum is dominant species(22 isolates), accounting for 40% of the total number of isolates. The remainder belongs to Cladosporium, Meyerozyma, Rhodotorula, Microsphaeropsis, Penicillium, Aspergillus, Phoma, Candida, Pestalotiopsis, and Engyodontium. Besides, two ascomycete isolates are unidentified. Seven indicator bacteria including Staphylococcus aureus were selected for antibacterial activity test. By using agar diffusion method, the bacteriostatic effect of the fungal isolates was verified. The result demonstrated that three isolates showed antibacterial activity for at least an indicator bacteria.
Six different kinds of culture media are used to isolate and cultivate the fungi associated with the South China Sea sponge. The isolates obtained totaled 55, belonging to 22 species. The identification wasmade through morphological observation in combination with ITS-rDNA blast homology analysis. The dominant genus is Trichoderma(31 isolates), and Trichoderma harzianum is dominant species(22 isolates), accounting for 40% of the total number of isolates. The remainder belongs to Cladosporium, Meyerozyma, Rhodotorula, Microsphaeropsis, Penicillium, Aspergillus, Phoma, Candida, Pestalotiopsis, and Engyodontium. Besides, two ascomycete isolates are unidentified. Seven indicator bacteria including Staphylococcus aureus were selected for antibacterial activity test. By using agar diffusion method, the bacteriostatic effect of the fungal isolates was verified. The result demonstrated that three isolates showed antibacterial activity for at least an indicator bacteria.
引文
Bao J,Sun YL,Zhang XY,Han Z,Gao HC,He F,Qian PY,Qi SH,2013.Antifouling and antibacterial polyketides from marine gorgonian coral-associated fungus Penicillum sp.SCSGAF0023.The Journal of Antibiotics,66(4):219-223
Bao J,Zhang XY,Xu XY,He F,Nong XH,Qi SH,2013.New cyclic tetrapeptides and asteltoxins from gorgonian-derived fungus Aspergillus sp.SCSGAF 0076.Tetrahedron,69(9):2113-2117
Guo SS,Xu W,Alias SA,He GY,Luo ZH,2018.Diversity of cultivable fungi associated with four coral species from the South China Sea.Journal of Applied Oceanography,37(1):35-44(in Chinese)
He F,Bao J,Zhang XY,Tu ZC,Shi YM,Qi SH,2013.Asperterrestide A,a cytotoxic cyclic tetrapeptide from the marine-derived fungus Aspergillus terreus SCSGAF0162.Journal of Natural Products,76(6):1182-1186
Khamthong N,Rukachaisirikul V,Phongpaichit S,Preedanon S,Sakayaroj J,2012.Bioactive polyketides from the sea fan-derived fungus Penicillum citrinum PSU-F51.Tetrahedron,68(39):8245-8250
Li Q,Wang G,2009.Diversity of fungal isolates from three Hawaiian marine sponges.Microbiological Research,64(4):233-241
Liu N,Li YX,Li ZY,Zhong M,Lei XL,2014.Advances in the natural products from coral-associated fungi and their activites.Chinese Journal of Marine Drugs,33(4):73-84(in Chinese)
Nissimov J,Rosenberg E,Munn CB,2009.Antimicrobial properties of resident coral mucus bacteria of Oculina patagonica.FEMS Microbiology Letters,292(2):210-215
Reveillaud J,Maigenien L,Eren AM,Huber JA,Apprill A,Sogin ML,Vanreusel A,2014.Host-specificity among abundant and rare taxa in the sponge microbiome.The ISME Journal,8(6):1198-1209
Rosenberg E,Koren O,Reshef L,Efrony R,Zilber-Rosenberg I,2007.The role of microorganisms in coral health,disease and evolution.Nature Reviews Microbiology,5(5):355
Shao CL,Wu HX,Wang CY,Qing AL,Xu Y,Wei MY,Qian PY,Gu YC,Zheng CJ,She ZG,Lin YC,2011.Potent antifouling resorcylic acid lactones from the gorgonian-derived fungus Cochliobolus lunatus.Journal of Natural Products,74(4):629-633
Wang G,Li Q,Zhu P,2008.Phylogenetic diversity of culturable fungi associated with the Hawaiian sponges.Antonie van Leeuwenhoek,93(1-2):163-174
Wei MY,Li D,Shao CL,Deng DS,Wang CY,2013.(±)-Pestalachloride D,an antibacterial racemate of chlorinated benzophenone derivative from a soft coral-derived fungus Pestalotiopsis sp.Marine Drugs,11(4):1050-1060
Yu Z,Zhang B,Sun W,Zhang F,Li Z,2013.Phylogenetically diverse cultivable endozoic fungi in the South China Sea sponges and their potential in synthesizing bioactive natural products suggested by PKS gene and cytotoxic activity analysis.Fungal Diversity,58(1):127-141
Zhang YJ,Zhang S,Liu XZ,Wen HA,Wang M,2010.A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains.Letters in Applied Microbiology,52(1):114-118
Zhou K,2010.Polyketide synthasa(PKS)and non-ribosomal petide syntnase(NRPS)genes feom phylogenetically diverse cultivable fungi associated with the South China Sea sponges.Master Thesis,Shanghai Jiaotong University,Shanghai.1-92(in Chinese)
Zhou K,Zhang X,Zhang F,Li ZY,2011.Phylogenetically diverse cultivable fungal community and polyketide synthase(PKS),non-ribosomal peptide synthase(NRPS)genes associated with South China seasponges.Microbial Ecology,62(3):644-654
Zhuang Y,Teng X,Wang Y,Liu P,Wang H,Li J,Li G,Zhu W,2011.Cyclopeptides and polyketides from coral-associated fungus Aspergillus versicolor LCJ-5-4.Tetrahedron,67(37):7085-7089
郭双双,徐炜,Siti Aisyah Alias,何高阳,骆祝华,2018.南海4种珊瑚附生可培养真菌多样性分析.应用海洋学学报,37(1):35-44
刘楠,李英新,李志勇,钟敏,雷晓淩,2014.珊瑚来源真菌天然产物及其活性的研究进展.中国海洋药物,33(4):73-84
周康,2010.中国南海海绵可培养共生真菌种群多样性、抑菌活性的研究及PK,NRPS功能基因的筛选.上海交通大学硕士学位论文,上海.1-92
Bao J,Zhang XY,Xu XY,He F,Nong XH,Qi SH,2013.New cyclic tetrapeptides and asteltoxins from gorgonian-derived fungus Aspergillus sp.SCSGAF 0076.Tetrahedron,69(9):2113-2117
Guo SS,Xu W,Alias SA,He GY,Luo ZH,2018.Diversity of cultivable fungi associated with four coral species from the South China Sea.Journal of Applied Oceanography,37(1):35-44(in Chinese)
He F,Bao J,Zhang XY,Tu ZC,Shi YM,Qi SH,2013.Asperterrestide A,a cytotoxic cyclic tetrapeptide from the marine-derived fungus Aspergillus terreus SCSGAF0162.Journal of Natural Products,76(6):1182-1186
Khamthong N,Rukachaisirikul V,Phongpaichit S,Preedanon S,Sakayaroj J,2012.Bioactive polyketides from the sea fan-derived fungus Penicillum citrinum PSU-F51.Tetrahedron,68(39):8245-8250
Li Q,Wang G,2009.Diversity of fungal isolates from three Hawaiian marine sponges.Microbiological Research,64(4):233-241
Liu N,Li YX,Li ZY,Zhong M,Lei XL,2014.Advances in the natural products from coral-associated fungi and their activites.Chinese Journal of Marine Drugs,33(4):73-84(in Chinese)
Nissimov J,Rosenberg E,Munn CB,2009.Antimicrobial properties of resident coral mucus bacteria of Oculina patagonica.FEMS Microbiology Letters,292(2):210-215
Reveillaud J,Maigenien L,Eren AM,Huber JA,Apprill A,Sogin ML,Vanreusel A,2014.Host-specificity among abundant and rare taxa in the sponge microbiome.The ISME Journal,8(6):1198-1209
Rosenberg E,Koren O,Reshef L,Efrony R,Zilber-Rosenberg I,2007.The role of microorganisms in coral health,disease and evolution.Nature Reviews Microbiology,5(5):355
Shao CL,Wu HX,Wang CY,Qing AL,Xu Y,Wei MY,Qian PY,Gu YC,Zheng CJ,She ZG,Lin YC,2011.Potent antifouling resorcylic acid lactones from the gorgonian-derived fungus Cochliobolus lunatus.Journal of Natural Products,74(4):629-633
Wang G,Li Q,Zhu P,2008.Phylogenetic diversity of culturable fungi associated with the Hawaiian sponges.Antonie van Leeuwenhoek,93(1-2):163-174
Wei MY,Li D,Shao CL,Deng DS,Wang CY,2013.(±)-Pestalachloride D,an antibacterial racemate of chlorinated benzophenone derivative from a soft coral-derived fungus Pestalotiopsis sp.Marine Drugs,11(4):1050-1060
Yu Z,Zhang B,Sun W,Zhang F,Li Z,2013.Phylogenetically diverse cultivable endozoic fungi in the South China Sea sponges and their potential in synthesizing bioactive natural products suggested by PKS gene and cytotoxic activity analysis.Fungal Diversity,58(1):127-141
Zhang YJ,Zhang S,Liu XZ,Wen HA,Wang M,2010.A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains.Letters in Applied Microbiology,52(1):114-118
Zhou K,2010.Polyketide synthasa(PKS)and non-ribosomal petide syntnase(NRPS)genes feom phylogenetically diverse cultivable fungi associated with the South China Sea sponges.Master Thesis,Shanghai Jiaotong University,Shanghai.1-92(in Chinese)
Zhou K,Zhang X,Zhang F,Li ZY,2011.Phylogenetically diverse cultivable fungal community and polyketide synthase(PKS),non-ribosomal peptide synthase(NRPS)genes associated with South China seasponges.Microbial Ecology,62(3):644-654
Zhuang Y,Teng X,Wang Y,Liu P,Wang H,Li J,Li G,Zhu W,2011.Cyclopeptides and polyketides from coral-associated fungus Aspergillus versicolor LCJ-5-4.Tetrahedron,67(37):7085-7089
郭双双,徐炜,Siti Aisyah Alias,何高阳,骆祝华,2018.南海4种珊瑚附生可培养真菌多样性分析.应用海洋学学报,37(1):35-44
刘楠,李英新,李志勇,钟敏,雷晓淩,2014.珊瑚来源真菌天然产物及其活性的研究进展.中国海洋药物,33(4):73-84
周康,2010.中国南海海绵可培养共生真菌种群多样性、抑菌活性的研究及PK,NRPS功能基因的筛选.上海交通大学硕士学位论文,上海.1-92