龙脑樟树叶内生真菌的分离及生物活性菌株的筛选鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2018-0750

生物技术通报 ›› 2019, Vol. 35 ›› Issue (3): 53-58.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0750

龙脑樟树叶内生真菌的分离及生物活性菌株的筛选鉴定

金宏杰^1,2, 曹红², 刘红¹, 郑爽², 姜超²

1. 石河子大学生命科学学院,石河子 832000;
2. 嘉兴学院生物与化学工程学院,嘉兴 314000

收稿日期:2018-08-28 出版日期:2019-03-26 发布日期:2019-04-03
作者简介:金宏杰,男,硕士研究生,研究方向：生物学;E-mail：395894596@qq.com
基金资助:
国家自然基金项目（21266029）,浙江省自然科学基金项目（LY17B060011）

Isolation of Endophytic Fungi from Cinnamomum camphora Leaves,Screening and Identification of Biologically Active Strains

JIN Hong-jie^1,2, CAO Hong², LIU Hong¹, ZHENG Shuang², JIANG Chao²

1. College of Life Sciences,Shihezi University,Shehezi 832000;
2. College of Biological,Chemical Sciences and Engineering,Jiaxing University,Jiaxing 314000

Received:2018-08-28 Published:2019-03-26 Online:2019-04-03

摘要/Abstract

摘要： 旨在从龙脑樟树叶片中分离内生真菌并对其发酵产物的体外抑菌、抗肿瘤和抗氧化活性进行测定,筛选鉴定具有较强生物活性的菌株。对分离出的内生真菌,用K-b纸片法测定其抑菌活性,MTT法测定抗肿瘤活性,紫外分光光度法测定其抗氧化活性,ITS（Internal transcribed spacer）序列测序分析确定活性菌株的种属。结果显示,共从龙脑樟树叶片中分离得到内生真菌39株,其中具有对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、铜绿假单胞菌较强抑制活性的菌株7株。又从7株真菌中筛选出具有较强抗肿瘤活性和抗氧化活性的菌株ZS14,其对人乳腺癌细胞MCF-7抑制率达到73.00%,对DPPH清除率70.92%,对羟自由基清除率78.83%,过氧化氢清除率81.36%,据ITS测序结果将其命名为Diaporthales sp. ZS14。分离得到的菌株Diaporthales sp. ZS14具有较强的抑菌、抗肿瘤、抗氧化活性,可以作为药物开发的潜在来源进行进一步研究。

关键词: 内生真菌, 抗菌, 抗肿瘤, 抗氧化, 分离鉴定

Abstract: This work aims to isolate endophytic fungi from leaves of Cinnamomum camphora,to determine its anti-bacterial activity,anti-tumor activity and anti-oxidation activity in vitro,and to screen and identify strains with strong biological activity. On the isolated endophytic fungi,K-b disk method was used to determine its anti-bacterial activity,MTT assay to determine its anti-tumor activity,ultraviolet spectrophotometry to determine its anti-oxidation activity,and ITS（Internal transcribed spacer）to analyze and determine the species of active strains. Results showed that total 39 endophytic fungi were isolated from the leaves of C. camphora.Among them,there were 7 strains highly inhibiting Escherichia coli,Staphylococcus aureus,Bacillus subtilis and Pseudomonas aeruginosa. Further,the strain ZS14 with anti-tumor and anti-oxidation activities from the 7 strains was screened,its inhibition rate on human breast cancer cell MCF-7 reached 73.00%,clearance rate of DPPH was 70.92%,scavenging rate of hydroxyl radical was 78.83%,and clearance rate of hydrogen peroxide was 81.36%. With ITS sequencing results it was designated as Diaporthales sp. ZS14. In sum,the isolated strain Diaporthales sp. ZS14 had strong anti-bacterial,anti-tumor and anti-oxidation activities,and it can be used as potential sources for drug development in further study.

Key words: endophytic fungi, anti-bacterial, anti-tumor, anti-oxidation, isolation and identification

金宏杰, 曹红, 刘红, 郑爽, 姜超. 龙脑樟树叶内生真菌的分离及生物活性菌株的筛选鉴定[J]. 生物技术通报, 2019, 35(3): 53-58.

JIN Hong-jie, CAO Hong, LIU Hong, ZHENG Shuang, JIANG Chao. Isolation of Endophytic Fungi from Cinnamomum camphora Leaves,Screening and Identification of Biologically Active Strains[J]. Biotechnology Bulletin, 2019, 35(3): 53-58.

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龙脑樟树叶内生真菌的分离及生物活性菌株的筛选鉴定

Isolation of Endophytic Fungi from Cinnamomum camphora Leaves,Screening and Identification of Biologically Active Strains

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