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中国农学通报 ›› 2022, Vol. 38 ›› Issue (1): 75-81.doi: 10.11924/j.issn.1000-6850.casb2021-0117

所属专题: 生物技术 植物保护 园艺

• 林学·园艺·园林 • 上一篇    下一篇

药用植物内生真菌的分离及拮抗菌的筛选与鉴定

王占斌(), 周暄, 马鑫博, 王靖琳   

  1. 东北林业大学林学院,哈尔滨 150040
  • 收稿日期:2021-02-02 修回日期:2021-09-26 出版日期:2022-01-05 发布日期:2022-02-24
  • 作者简介:王占斌,男,黑龙江哈尔滨人,副教授,博士,研究方向:森林病理学。通信地址:150080 黑龙江省哈尔滨市和兴路26号,E-mail: wangzbchina@163.com
  • 基金资助:
    中央高校基本业务专项资金项目“水曲柳重大病虫害预警与防控新技术的研究”(2572020DR09)

Isolation of Endophytic Fungi from Medicinal Plants and Screening and Identification of Antagonism

WANG Zhanbin(), ZHOU Xuan, MA Xinbo, WANG Jinglin   

  1. College of Forestry, Northeast Forestry University, Harbin 150040
  • Received:2021-02-02 Revised:2021-09-26 Online:2022-01-05 Published:2022-02-24

摘要:

为更好地研究药用植物内生拮抗菌种,以6种药用植物(连钱草、小白酒草、毛茛、土三七、唐松草、白屈菜)作为植物材料进行菌种提取分离,并检测其抑菌活性。选择其中3种抑菌活性较高菌株通过18S rDNA序列测定,构建系统发育树,进行分子生物学鉴定。结果表明,6种药用植物均含有多种内生真菌,连钱草分离得到5株内生真菌,小白酒草2株,毛茛4株,土三七2株,唐松草1株,白屈菜2株。共获得对杨树叶枯病原真菌抑制作用较强的7株拮抗菌,分别为LJ3、MJ2、MY1、TJ1、TJ2、J1、BY1。其中MJ2对杨叶枯病的抑制率最高,可达56.47%。经鉴定MY1与BY1为粉红粘帚霉(Clonostachys rosea),TJ2为镰孢霉属(Fusarium)的一个菌株。抑菌试验认为,本试验药用植物内生菌对杨树叶枯病具有比较明显的拮抗作用,可用于植物病害的生物防治。

关键词: 药用植物, 内生真菌, 分离培养, ITS, 鉴定, 拮抗

Abstract:

To study the endophytic antagonistic strains, six medicinal plants, including Glechoma longituba, Conyza canadensis, Ranunculus japonicus, Gynura segetum, Thalictrum aquilegifolium and Chelidonium majus, were used as plant materials to extract and isolate the strains, and the antibacterial activities were tested. Three strains with high antibacterial activity were selected for molecular identification through 18S rDNA sequencing and phylogenetic tree construction. The results showed that all the six medicinal plants contained endophytic fungi, and 5 endophytic fungi were isolated from Glechoma longituba, 2 from Conyza canadensis, 4 from Ranunculus japonicus, 2 from Gynura segetum, 1 from Thalictrum aquilegifolium, and 2 from Chelidonium majus. Seven strains of antagonistic bacteria were obtained, which were LJ3, MJ2, MY1, TJ1, TJ2, J1 and BY1. MJ2 had the highest inhibitory rate of 56.47% against Alternaria alternata. MY1 and BY1 were identified as Clonostachys rosea, and TJ2 as a strain of Fusarium. It is proved that the endophytic bacteria have obvious antagonistic effect on Alternaria alternata and could be used for biological control of plant diseases.

Key words: medicinal plants, endophytic fungi, isolation and culture, ITS, identification, antagonism

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