刺五加内生真菌分离鉴定及抗菌活性研究
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摘要
为开发利用刺五加内生真菌资源,寻找新型抗菌物质,从刺五加根茎叶中分离纯化得到内生真菌71株,经菌落形态和显微形态去重后进行ITS序列分子鉴定为18个种,来自于4目、8科、11属,其中链格孢属与镰孢霉属为优势属。根、茎、叶中内生真菌多样性指数分别为1. 112 31、1. 523 658、0. 264 1。18种刺五加内生真菌中,16种内生真菌至少对一种指示菌有抗菌活性。CWJ-5(镰孢霉属Fusarium oxysporum)对金黄色葡萄球菌抑菌效果显著,抑菌直径27. 17±0. 07 mm; CWJ-6(链格孢属Alternaria alternata)对大肠埃希菌抑菌效果显著,抑菌直径18. 07±0. 01 mm。结果表明刺五加内生真菌资源丰富,有很大的开发利用空间,部分菌种抑菌活性显著,值得进一步研究。
In order to develop and use the endophytic fungi resources of Acanthopanax senticosus( Rupr. Maxim.) Harms and to find novel antibacterial substances,71 strains of endophytic fungi were isolated and purified from the roots,stems and leaves of Acanthopanax senticosus( Rupr. Maxim.) Harms. Molecular identification of ITS sequences after decontamination by colony morphology and microscopic morphology,we got 18 species from 4 orders,8 families and 11 genuses,of which Alternaria and Fusarium are dominant genuses. The endophytic fungi diversity index in roots,stems and leaves were 1. 123 31,1. 523658,0. 264 1,respectively. Among the 18 species of endophytic fungi of Acanthopanax senticosus( Rupr. Maxim.) Harms,16 endophytic fungi have antibacterial activity against at least one indicator bacteria. CWJ-5( Fusarium oxysporum) has asignificant antibacterial effect against Staphylococcus aureus,with a bacteriostasis diameter of 27. 17 ± 0. 07 mm; CWJ-6( Alternaria alternata) has significant antibacterial effect against Escherichia coli.,and its antibacterial diameter can reach 18. 07 ± 0. 01 mm. The results showed that the endophytic fungi of Acanthopanax senticosus( Rupr. Maxim.) Harms was rich in resources,and there was a great deal of space for development and utilization. Some of the strains had significant antibacterial activity and deserved further study.
In order to develop and use the endophytic fungi resources of Acanthopanax senticosus( Rupr. Maxim.) Harms and to find novel antibacterial substances,71 strains of endophytic fungi were isolated and purified from the roots,stems and leaves of Acanthopanax senticosus( Rupr. Maxim.) Harms. Molecular identification of ITS sequences after decontamination by colony morphology and microscopic morphology,we got 18 species from 4 orders,8 families and 11 genuses,of which Alternaria and Fusarium are dominant genuses. The endophytic fungi diversity index in roots,stems and leaves were 1. 123 31,1. 523658,0. 264 1,respectively. Among the 18 species of endophytic fungi of Acanthopanax senticosus( Rupr. Maxim.) Harms,16 endophytic fungi have antibacterial activity against at least one indicator bacteria. CWJ-5( Fusarium oxysporum) has asignificant antibacterial effect against Staphylococcus aureus,with a bacteriostasis diameter of 27. 17 ± 0. 07 mm; CWJ-6( Alternaria alternata) has significant antibacterial effect against Escherichia coli.,and its antibacterial diameter can reach 18. 07 ± 0. 01 mm. The results showed that the endophytic fungi of Acanthopanax senticosus( Rupr. Maxim.) Harms was rich in resources,and there was a great deal of space for development and utilization. Some of the strains had significant antibacterial activity and deserved further study.
引文
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15 Chen HJ.Isolation and identification of endophytic fungi from wild Acanthopanax[J].J Pract Tradit Chin Med(实用中医药杂志),2016,32:740-741.
16 Liu J,Gao Y,Luo JH.Isolation and identification of endophytic fungi from wild Acanthopanax Senticosus[J].Heilongjiang Med and Pharm(黑龙江医药科学),2011,34(4):8-9.
17 Xiong YN,Xing ZB,Wu P,et al.Study on isolation and distribution of endophytic fungus in Eleutherococcus senticosus Harms[J].J Anhui Agric Sci(安徽农业科学),2009,37:11347-11348.
18 Lyu LX,Wang HW,Liang XF,et al.Effects of different chemotypes and seasonal dynamic variation on the species diversity of endophytic fungal communities harbored in Atractylodes lancea[J].Acta Ecol Sin(生态学报),2014,34:7300-7310.
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20 Li ZY,Chen YW,Yang LY,et al.Antipathogenic activity of endophytic fungi in Radixaconiti plant[J].Nat Prod Res Dev(天然产物研究与开发),2009,21:676-678.
21 Meng SX,Cao J,Zhang HR,et al.Antibacterial mechanisms of endophytic fungi from Gynostemma pentaphyllum on Staphylococcus aureus[J].J Chin Antibiotics(中国抗生素杂志),2015,40:268-273.
2 Cui JL,Guo SX,Xiao PG.Interaction between endophytes and host plant and the role of endophytes in genuineness analysis of medicinal plant[J].Acta Pharm Sin(药学学报),2017,52:214-221.
3 Stone JK,Bacon CW,White JF.An overview of endophytic microbes:endophytism defined,3[M].New York:Marcel Dekker,2000:267-274.
4 Guo SX.The recent progress and prospects of research on endophytic fungi in medicinal plants[J].Mycosystema(菌物学报),2018,37(1):1-13.
5 Qiu DY,Huang MJ,Fang XH,et al.Isolation of an endophytic fungus associated taxus yunnanensis et L.K.Fu[J].Aeto Myeologiea Sin(真菌学报),1994,4:314-316.
6 Li J),Zhang HR,Liu NY,et al.Study on isolation and identification endophytic fungi and antibacterial activity of Flos Lonicerae[J].J Chin Antibiotics(中国抗生素杂志),2010,35:236-237.
7 Zhu YG,Ouyang WY,Wu N,et al.Antibiotic resistance sources and mitigation[J].Bull Chin Acad Sci(中国科学院院刊),2015,30:509-516.
8 Shi QY,Chen XM,Guo SX.Research progress of antibacterial active substance derived from endophytic fungi from plants[J].Chin Pharm J(中国药学杂志)2007,11:804-807.
9 Chinese Pharmacopoeia Commission(国家药典委员会).Pharmacopoeia of the People’s Republic of China:VolⅠ(中华人民共和国药典:第一部)[M].Beijing:China Medical Science Press,2015:206-207.
10 Yang XD,Jing YE,Lu F.Research progress on chemical components of Acanthopanax senticosus[J].Chin Arcs Tradit Chin Med(中华中医药学刊),2015,33:316-318.
11 Bai X,Hu WZ,Jiang AL,et al.Research advance on main active substance and pharmacological action of Acanthopanax[J].Sci Tech Food Ind(食品工业科技),2016,37:396-399.
12 Huang J,Shao Q,Xiang YH,et al.Identification of phenylpropanoids in Ciwujia injection by HPLC-MS[J].Chin JChin Mater Med(中国中药杂志),2014,39:2513-2520.
13 Zheng P,Tan DG,Sun PX,et al.Morphology and phylogenetic position of an endophytic fungus ITBB2-1 from rubber tree[J].Chin J Trop Crops(热带作物学报),2009,30:314-319.
14 Pielou EC.Ecological diversity[M].New York:John Wiley and Sons Inc,1975.
15 Chen HJ.Isolation and identification of endophytic fungi from wild Acanthopanax[J].J Pract Tradit Chin Med(实用中医药杂志),2016,32:740-741.
16 Liu J,Gao Y,Luo JH.Isolation and identification of endophytic fungi from wild Acanthopanax Senticosus[J].Heilongjiang Med and Pharm(黑龙江医药科学),2011,34(4):8-9.
17 Xiong YN,Xing ZB,Wu P,et al.Study on isolation and distribution of endophytic fungus in Eleutherococcus senticosus Harms[J].J Anhui Agric Sci(安徽农业科学),2009,37:11347-11348.
18 Lyu LX,Wang HW,Liang XF,et al.Effects of different chemotypes and seasonal dynamic variation on the species diversity of endophytic fungal communities harbored in Atractylodes lancea[J].Acta Ecol Sin(生态学报),2014,34:7300-7310.
19 Li JB,Ding CH,Zhang S,et al.Preliminary study on the growth-promoting effect of endophytic fungi from rumex gmelini turcz[J].Inf Tradit Chin Med(中医药信息),2018,35(1):6-9.
20 Li ZY,Chen YW,Yang LY,et al.Antipathogenic activity of endophytic fungi in Radixaconiti plant[J].Nat Prod Res Dev(天然产物研究与开发),2009,21:676-678.
21 Meng SX,Cao J,Zhang HR,et al.Antibacterial mechanisms of endophytic fungi from Gynostemma pentaphyllum on Staphylococcus aureus[J].J Chin Antibiotics(中国抗生素杂志),2015,40:268-273.