樟叶越桔1株内生真菌的鉴定及其抑菌活性
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摘要
从野生樟叶越桔叶中分离得到1株内生真菌VDL117,利用形态学分类方法、ITS序列测定以及系统发育分析鉴定其为曲霉属杂色曲霉。采用生长速率法测定了其发酵液对棉花枯萎菌等5种常见农作物病原真菌的抑菌活性,并对其发酵液进行了酸碱稳定性及热稳定性实验。结果表明:VDL117发酵液对5种病原真菌均有一定的抑制作用,发酵液在酸性条件下对5种病原真菌的抑制活性较强,碱性条件下活性减弱,并在70℃及以下温度能保持活性稳定。
An endophytic fungus,strain VDL117,was isolated from the leaves of wild Vaccinium dunalianum,and identified as Aspergillus versicolor by morphological classification method,ITS sequences and phylogenetic analysis. Using the growth rate method,the antifungal activity of its fermentation broth on 5 plant pathogenic fungi was tested on different temperature and p H conditions. The results show that the fermentation broth of strain VDL117 has a certain inhibitory effects on 5 plant pathogenic fungi. The antifungal activity can be reduced or even lost under alkaline condition,while acidic condition can enhance its inhibitory effect. The activity is stable at 70 ℃ and below the temperature.
An endophytic fungus,strain VDL117,was isolated from the leaves of wild Vaccinium dunalianum,and identified as Aspergillus versicolor by morphological classification method,ITS sequences and phylogenetic analysis. Using the growth rate method,the antifungal activity of its fermentation broth on 5 plant pathogenic fungi was tested on different temperature and p H conditions. The results show that the fermentation broth of strain VDL117 has a certain inhibitory effects on 5 plant pathogenic fungi. The antifungal activity can be reduced or even lost under alkaline condition,while acidic condition can enhance its inhibitory effect. The activity is stable at 70 ℃ and below the temperature.
引文
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[33]罗寒,李晓栋,李晓明,等.红树林来源内生真菌杂色曲霉Aspergillus versicolor MA-229次级代谢产物研究[J].中国抗生素杂志,2017,42(4):334-340.
[34]吴亚腾,吴曼莉,柳志强,等.大叶山楝拮抗内生真菌DYSJ3的鉴定及抑菌活性产物分析[J].江苏农业科学,2016,45(5):181-184.
[35]伊艳杰,时玉,吴兴泉,等.禾谷丝核菌拮抗真菌的鉴定及抗菌活性研究[J].河南农业科学,2010,39(8):86-89.
[36]董世豪,巩婷,朱平.相似蜂海绵相关真菌杂色曲霉F62活性代谢产物研究[J].菌物学报,2011,30(4):636-643.
[37]汪茜,胡春锦,柯仿钢,等.生防菌株1404的鉴定及其对采后柑橘炭疽病的防治效果[J].微生物学报,2010,50(9):1208-1217.
[2]Stierle A,Strobel G,Stierle D.Taxol and taxane production by Taxomyces andreanae,an endophytic fungus of Pacific yew[J].Science,1993,260(5105):214-216.
[3]Stierle A,Strobel G,Stierle D,et al.The search for a taxol-producing microorganism among the endophytic fungi of the Pacific yew,Taxus brevifolia[J].Journal of Natural Products,1995,58(9):1315-1324.
[4]Puri S C,Verma V,Amna T,et al.An endophytic fungus from Nothapodytes foetida that produces camptothecin[J].Journal of Natural Products,2005,68(12):1717-1719.
[5]Wang X,Radwan M M,Taráwneh A H,et al.Antifungal activity against plant pathogens of metabolites from the endophytic fungus Cladosporium cladosporioides[J].Journal of Agricultural&Food Chemistry,2013,61(19):4551-4555.
[6]易晓华.植物内生真菌次生代谢产物研究进展[J].中国农学通报,2009,25(21):255-260.
[7]Kumar S,Kaushik N.Metabolites of endophytic fungi as novel source of biofungicide:a review[J].Phytochemistry Reviews,2012,11(4):507-522.
[8]方瑞征.中国越桔属的研究[J].云南植物研究,1986,8(3):239-258.
[9]夏燕,赵平,刘云.彝族雀嘴茶饮料的研制[J].食品研究与开发,2015,36(9):76-80.
[10]Zhao P,Tanaka T,Hirabayashi K,et al.Caffeoyl arbutin and related compounds from the buds of Vaccinium dunalianum[J].Phytochemistry,2008,69(18):3087-3094.
[11]阮珍美,陈火根.高丛越桔内生真菌的分离及其优势种群的鉴定[J].现代园艺,2010,33(5):9-10.
[12]李亚东,张大鹏.越桔菌根真菌研究进展:文献综述[J].园艺学报,1996,23(2):133-138.
[13]李姝诺,李亚东,王琦.越橘产黄酮类化合物内生真菌的筛选[J].吉林农业大学学报,2009,31(5):587-591.
[14]苗迎秋,王贺新,李根柱,等.长白山笃斯越橘菌根形态结构及内生菌的分布特征[J].东北农业大学学报,2013,44(1):81-85.
[15]刘凤红,程显好,顾亮,等.长白山野生笃斯越橘根系内生菌资源调查[J].山东农业科学,2015,47(1):41-46.
[16]白永超,陈露,卫旭芳,等.大兴安岭笃斯越橘内生真菌及矿质养分特性分析[J].林业科学,2017,53(10):50-59.
[17]Li Z J,Shen X Y,Hou C L.Fungal endophytes of South China blueberry(Vaccinium dunalianum var.urophyllum)[J].Letters in Applied Microbiology,2016,63(6):482-487.
[18]Baba T,Hirose D,Sasaki N,et al.Mycorrhizal formation and diversity of endophytic fungi in hair roots of Vaccinium oldhamii Miq.in Japan[J].Microbes&Environments,2016,31(2):186-189.
[19]Li P Q,Wu Z,Liu T,et al.Biodiversity,phylogeny,and antifungal functions of endophytic fungi associated with Zanthoxylum bungeanum[J].International Journal of Molecular Sciences,2016,17(9):1541.
[20]张艳军.番茄早疫病菌拮抗菌发酵液稳定性分析[J].生物技术,2012,22(6):74-77.
[21]齐祖同.中国真菌志:第5卷[M].北京:科学出版社,1997.
[22]Siqueira J P,Sutton D A,García D,et al.Species diversity of Aspergillus section Versicolores in clinical samples and antifungal susceptibility[J].Fungal Biology,2016,120(11):1458-1467.
[23]Souza P N,Grigoletto T L,de Moraes L A,et al.Production and chemical characterization of pigments in filamentous fungi[J].Microbiology,2016,162(1):12-22.
[24]Samson R A,Varga J,Meijer M,et al.New taxa in Aspergillus,section Usti[J].Studies in Mycology,2011,69(1):81-97.
[25]Hajji M,Kanoun S,Nasri M,et al.Purification and characterization of an alkaline serine-protease produced by a new isolated Aspergillus clavatus ES1[J].Process Biochemistry,2007,42(5):791-797.
[26]You Y H,Kwak T W,Kang S M,et al.Aspergillus clavatus Y2H0002 as a new endophytic fungal strain producing gibberellins isolated from Nymphoides peltata in fresh water[J].Mycobiology,2015,43(1):87-91.
[27]Accinelli C,Mencarelli M,SaccàM L,et al.Managing and monitoring of Aspergillus flavus,in corn using bioplastic-based formulations[J].Crop Protection,2012,32(1358):30-35.
[28]Bajaj B K,Abbass M.Studies on an alkali-thermostable xylanase from Aspergillus fumigatus MA28[J].Biotech,2011,1(3):161-171.
[29]Chen Y,Wang H,Lu Z,et al.Emergence of TR46/Y121F/T289A in an Aspergillus fumigatus isolate from a Chinese patient[J].Antimicrobial Agents&Chemotherapy,2015,59(11):7148-7150.
[30]Paavanenhuhtala S,Hyvonen J,Bulat S A,et al.RAPDPCR,isoenzyme,r DNA RFLP and r DNA sequence analyses in identification of Finnish Fusarium oxysporum isolates[J].Mycological Research,1999,103(5):625-634.
[31]Wang Y,Qi S,Zhan Y,et al.Aspertetranones A-D,putative meroterpenoids from the marine algal-associated fungus Aspergillus sp.ZL0-1b14[J].Journal of Natural Products,2015,78(10):2405-2410.
[32]Xu J,Zhao S and Yang X,A new cyclopeptide metabolite of marine gut fungus from Ligia oceanic[J].Natural Product Research,2014,28(13):994-997.
[33]罗寒,李晓栋,李晓明,等.红树林来源内生真菌杂色曲霉Aspergillus versicolor MA-229次级代谢产物研究[J].中国抗生素杂志,2017,42(4):334-340.
[34]吴亚腾,吴曼莉,柳志强,等.大叶山楝拮抗内生真菌DYSJ3的鉴定及抑菌活性产物分析[J].江苏农业科学,2016,45(5):181-184.
[35]伊艳杰,时玉,吴兴泉,等.禾谷丝核菌拮抗真菌的鉴定及抗菌活性研究[J].河南农业科学,2010,39(8):86-89.
[36]董世豪,巩婷,朱平.相似蜂海绵相关真菌杂色曲霉F62活性代谢产物研究[J].菌物学报,2011,30(4):636-643.
[37]汪茜,胡春锦,柯仿钢,等.生防菌株1404的鉴定及其对采后柑橘炭疽病的防治效果[J].微生物学报,2010,50(9):1208-1217.