蜂巢珊瑚共生真菌的分离、鉴定及其抗氧化活性
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摘要
为从蜂巢珊瑚中寻找具有高抗氧化活性的共生真菌,采用组织研磨法分离蜂巢珊瑚共生真菌,通过对DPPH、ABTS自由基清除及对高价铁离子的还原能力的测定筛选高抗氧化活性的菌株,通过形态学特征观察,18S rDNA序列分析对高抗氧化活性菌株进行鉴定。从蜂巢珊瑚中共分离9株共生真菌;抗氧化活性测定结果表明,9株真菌发酵液均具有较好的抗氧化活性,其中S-1-5菌株的抗氧化活性最好,其对DPPH和ABTS自由基清除率分别为70.60%和67.18%,对高价铁离子还原能力强。依据S-1-5菌株的形态学特征和18S rDNA序列分析结果,将菌株S-1-5鉴定为球孢枝孢(Cladosporium sphaerospermum)。该菌株具有进一步开发应用的潜在价值。
To screen fungi with high antioxidant activity from Favia speciosa,epiphyte fungi were isolated from Favia speciosa by tissue grinding method.DPPH free radical scavenging capacity,ABTS free radical scavenging capacity and ferric iron reduction capacity were determined.Then,the morphological characteristics and 18 S rDNA sequence analysis of screened isolates was identified.The result showed that nine kinds of fungi were isolated from Favia speciosa.The fermentation broth of 9 strains had good antioxidant activity,especially,S-1-5 strain represented the best antioxidant activity among the strains,with 70.60% of DPPH free radical scavenging rate and 67.18% of ABTS free radical scavenging rate.Based on the morphology and 18 S rDNA sequence, S-1-5 strain was identified as Cladosporium sphaerospermum,which has a potential application value.
To screen fungi with high antioxidant activity from Favia speciosa,epiphyte fungi were isolated from Favia speciosa by tissue grinding method.DPPH free radical scavenging capacity,ABTS free radical scavenging capacity and ferric iron reduction capacity were determined.Then,the morphological characteristics and 18 S rDNA sequence analysis of screened isolates was identified.The result showed that nine kinds of fungi were isolated from Favia speciosa.The fermentation broth of 9 strains had good antioxidant activity,especially,S-1-5 strain represented the best antioxidant activity among the strains,with 70.60% of DPPH free radical scavenging rate and 67.18% of ABTS free radical scavenging rate.Based on the morphology and 18 S rDNA sequence, S-1-5 strain was identified as Cladosporium sphaerospermum,which has a potential application value.
引文
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[13] DU B,XU B.Oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) of β-glucans from different sources with various molecular weight [J].Bioactive Carbohydrates and Dietary Fibre,2014,3(1):11-16.
[14] BOUAZIZ M,YANGUI T,SAYADI S,et al.Disinfectant properties of essential oils from Salvia officinalis L.cultivated in Tunisia [J].Food and Chemical Toxicology,2009,47(11):2755-2760.
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[17] 张先富,相立,王艳芳,等.草酸青霉A1菌株的鉴定及对苹果4种镰孢病菌的拮抗作用 [J].园艺学报,2016,43(5):841-852. ZHANG X F,XIANG L,WANG Y F,et al.Identification of Penicillium oxalicum A1 strain and antagonistic effects on four species of Fusarium pathogen of apple [J].Acta Horticulturae Sinica,2016,43(5):841-852.
[18] 楚文卉,谢清哲,杨超,等.1株镉耐受真菌C12的鉴定及其镉耐受性研究 [J].西北农林科技大学学报(自然科学版),2017,45(9):71-80. CHU W H,XIE Q ZH,YANG CH,et al.The identification and study on Cd tolerance of C12,a fungus which shows high tolerance to Cd [J].Journal of Northwest A&F University(Natural Science Edition),2017,45(9):71-80.
[19] ZALAR P,DE HOOG G S,SCHROERS H J,et al.Phylogeny and ecology of the ubiquitous saprobe Cladosporium sphaerospermum,with descriptions of seven new species from hypersaline environments [J].Studies in Mycology,2007,58:157-183.
[20] 刘玲枝,黄惠琴,鲍时翔.海绵共附生微生物的研究新进展 [J].生物技术通报,2007(3):93-96. LIU L ZH,HUANG H Q,BAO SH X.Progress in studies on sponge-associated microorganism [J].Biotechnology Bulletin,2007(3):93-96.
[21] RADJASA O K,VASKE Y M,NAVARRO G,et al.Highlights of marine invertebrate-derived biosynthetic products:their biomedical potential and possible production by microbial associants [J].Bioorganic & Medicinal Chemistry,2011,19(22):6658-6674.
[22] ROSENBERG E,KOREN O,RESHEF L,et al.The role of microorganisms in coral health,disease and evolution [J].Nature Reviews Microbiology,2007,5(5):355-362.
[23] MOU X F,LIU X,XU R F,et al.Scopuquinolone B,a new monoterpenoid dihydroquinolin-2 (1 H)-one isolated from the coral-derived Scopulariopsis sp.fungus [J].Natural Product Research,2018,32(7):773-776.
[24] ELNGGGAR M S,EBADA S S,ASHOUR M L,et al.Two new triterpenoids and a new naphthoquinone derivative isolated from a hard coral-derived fungus Scopulariopsis sp.[J].Fitoterapia,2017,116:126-130.
[25] XU W,GUO S,GONG L,et al.Phylogenetic survey and antimicrobial activity of cultivable fungi associated with five scleractinian coral species in the South China Sea [J].Botanica Marina,2018,61(1):75-84.
[26] ZHENG C J,SHAO C L,GUO Z Y,et al.Bioactive hydroanthraquinones and anthraquinone dimers from a soft coral-derived Alternaria sp.fungus [J].Journal of Natural Products,2012,75(2):189-197.
[27] 蔡隽妮.南海石珊瑚Diploria strigosa共生微生物活性次级代谢产物的研究 [D].广州:广东药学院,2012. CAI J N.Study on the bioactive secondary metabolites of symbiotic microorganism from stony coral Diploria strigosa of the South China Sea [D].Guangzhou:Guangdong Pharmaceutical University,2012.
[28] 陈艳丽.不同海洋生境来源微生物胞外多糖的结构及抗氧化活性研究[D].山东青岛:中国海洋大学,2013. CHEN Y L.Study on the structure and antioxidant activity of exopolysaccharide from different marine habitats[D].Qingdao Shandong:Ocean University of China,2013.
[2] SUN C,WANG J W,FANG L,et al.Free radical scavenging and antioxidant activities of EPS2,an exopolysaccharide produced by a marine filamentous fungus Keisslerella sp.4108 [J].Life Sciences,2004,75:1063-1073.
[3] 阎雪芬,黄丹虹,代宇博,等.海洋真菌抗氧化活性物质的初步研究 [J].厦门大学学报(自然科学版),2007,46(3):418-421. YAN X F,HUANG D H,DAI Y B,et al.Preliminary study on antioxidant substances from marine fungi [J].Journal of Xiamen University(Natural Science Edition),2007,46(3):418-421.
[4] SUN H H,MAO W J,CHEN Y,et al.Isolation,chemical characteristics and antioxidant properties of the polysaccharides from marine fungus Penicillium sp.F23-2 [J].Carbohydrate Polymers,2009,78(1):117-124.
[5] CROFT M T,LAWRENCE A D,RAUX-DEERY E,et al.Algae acquire vitamin B12 through a symbiotic relationship with bacteria [J].Nature,2005,438(7064):90-93.
[6] 刘楠,李英新,李志勇,等.珊瑚来源真菌天然产物及其活性的研究进展 [J].中国海洋药物,2014,33(4):73-84. LIU N,LI Y X,LI ZH Y,et al.Advances in the natural products from coral-associated fungi and their activities [J].Chinese Journal of Marine Drugs,2014,33(4):73-84.
[7] 姚远蓓,班海侠,雷晓凌,等.徐闻普哥滨珊瑚土曲霉XWC21-10代谢产物活性及其活性物质的初步研究 [J].微生物学杂志,2017,37(6):18-23. YAO Y B,BAN H X,LEI X L,et al.Preliminarily study on metabolites activity and bioactive substances of Aspergillus terreus XWC21-10 from Porites pukoensis in Xuwen [J].Journal of Microbiology,2017,37(6):18-23.
[8] 李耀西.蜂巢珊瑚类横板珊瑚的体壁构成及有关问题 [J].古生物学报,1991,30(2):231-238. LI Y X.On formation of the wall of favositids(tabulata) and other related problems [J].Acta Palaeontologica Sinica,1991,30(2):231-238.
[9] 刘苗苗,沈建伟,王月,等.雷州半岛徐闻西岸珊瑚岸礁造礁珊瑚群落结构及其演变 [J].海洋地质与第四纪地质,2011,31(6):37-45. LIU M M,SHEN J W,WANG Y,et al.Reef-building coral community and evolutionary characteristics of the fringing reefs on the west coast of Xuwen County,Leizhou Peninsula [J].Marine Geology & Quaternary Geology,2011,36 (6):37-45.
[10] 佚名.广东用角蜂巢珊瑚为原料制成人工骨 [J].医学信息(上旬刊),1997,10(7):8. YI M.Using horned hive coral as raw material to make artificial bone in Guangdong [J].Medical Information (First-10 Day Edition),1997,10(7):8.
[11] 肖胜蓝,雷晓凌,徐佳,等.徐闻8种珊瑚共附生真菌的分离及初步鉴定 [J].广东海洋大学学报,2011,31(6):91-95. XIAO SH L,LEI X L,XU J,et al.Isolation and identification of fungus associated with eight corals from Xuwen [J].Journal of Guangdong Ocean University,2011,31(6):91-95.
[12] 陈志敏,田小曼,吴云锋.火棘内生真菌的分离及抑菌活性测定 [J].西北农业学报,2010,19(1):64-67. CHEN ZH M,TIAN X M,WU Y F.Isolation of endophytes from Pyracantha fortuneana and the test on fungistasis of some endophytic fungi [J].Acta Agriculturae Boreali-occidentalis Sinica,2010,19(1):64-67.
[13] DU B,XU B.Oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) of β-glucans from different sources with various molecular weight [J].Bioactive Carbohydrates and Dietary Fibre,2014,3(1):11-16.
[14] BOUAZIZ M,YANGUI T,SAYADI S,et al.Disinfectant properties of essential oils from Salvia officinalis L.cultivated in Tunisia [J].Food and Chemical Toxicology,2009,47(11):2755-2760.
[15] THOO Y Y,HO S K,LIANG J Y,et al.Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu(Morinda citrifolia) [J].Food Chemistry,2010,120(1):290-295.
[16] 宋新月,汤冰雪,邱君志,等.竹叶兰内生真菌的分离鉴定及其抗氧化活性研究 [J].现代食品科技,2018,34(2):1-6 SONG X Y,TANG B X,QIU J ZH,et al.Isolation,identification and antioxidant activity of endophytes in Arundia graminifolia [J].Modern Food Science and Technology,2018,34 (2):1-6.
[17] 张先富,相立,王艳芳,等.草酸青霉A1菌株的鉴定及对苹果4种镰孢病菌的拮抗作用 [J].园艺学报,2016,43(5):841-852. ZHANG X F,XIANG L,WANG Y F,et al.Identification of Penicillium oxalicum A1 strain and antagonistic effects on four species of Fusarium pathogen of apple [J].Acta Horticulturae Sinica,2016,43(5):841-852.
[18] 楚文卉,谢清哲,杨超,等.1株镉耐受真菌C12的鉴定及其镉耐受性研究 [J].西北农林科技大学学报(自然科学版),2017,45(9):71-80. CHU W H,XIE Q ZH,YANG CH,et al.The identification and study on Cd tolerance of C12,a fungus which shows high tolerance to Cd [J].Journal of Northwest A&F University(Natural Science Edition),2017,45(9):71-80.
[19] ZALAR P,DE HOOG G S,SCHROERS H J,et al.Phylogeny and ecology of the ubiquitous saprobe Cladosporium sphaerospermum,with descriptions of seven new species from hypersaline environments [J].Studies in Mycology,2007,58:157-183.
[20] 刘玲枝,黄惠琴,鲍时翔.海绵共附生微生物的研究新进展 [J].生物技术通报,2007(3):93-96. LIU L ZH,HUANG H Q,BAO SH X.Progress in studies on sponge-associated microorganism [J].Biotechnology Bulletin,2007(3):93-96.
[21] RADJASA O K,VASKE Y M,NAVARRO G,et al.Highlights of marine invertebrate-derived biosynthetic products:their biomedical potential and possible production by microbial associants [J].Bioorganic & Medicinal Chemistry,2011,19(22):6658-6674.
[22] ROSENBERG E,KOREN O,RESHEF L,et al.The role of microorganisms in coral health,disease and evolution [J].Nature Reviews Microbiology,2007,5(5):355-362.
[23] MOU X F,LIU X,XU R F,et al.Scopuquinolone B,a new monoterpenoid dihydroquinolin-2 (1 H)-one isolated from the coral-derived Scopulariopsis sp.fungus [J].Natural Product Research,2018,32(7):773-776.
[24] ELNGGGAR M S,EBADA S S,ASHOUR M L,et al.Two new triterpenoids and a new naphthoquinone derivative isolated from a hard coral-derived fungus Scopulariopsis sp.[J].Fitoterapia,2017,116:126-130.
[25] XU W,GUO S,GONG L,et al.Phylogenetic survey and antimicrobial activity of cultivable fungi associated with five scleractinian coral species in the South China Sea [J].Botanica Marina,2018,61(1):75-84.
[26] ZHENG C J,SHAO C L,GUO Z Y,et al.Bioactive hydroanthraquinones and anthraquinone dimers from a soft coral-derived Alternaria sp.fungus [J].Journal of Natural Products,2012,75(2):189-197.
[27] 蔡隽妮.南海石珊瑚Diploria strigosa共生微生物活性次级代谢产物的研究 [D].广州:广东药学院,2012. CAI J N.Study on the bioactive secondary metabolites of symbiotic microorganism from stony coral Diploria strigosa of the South China Sea [D].Guangzhou:Guangdong Pharmaceutical University,2012.
[28] 陈艳丽.不同海洋生境来源微生物胞外多糖的结构及抗氧化活性研究[D].山东青岛:中国海洋大学,2013. CHEN Y L.Study on the structure and antioxidant activity of exopolysaccharide from different marine habitats[D].Qingdao Shandong:Ocean University of China,2013.