天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (7): 1149-1154.doi: 10.16333/j.1001-6880.2018.7.009

• 研究论文 • 上一篇    下一篇

一株川贝母内生真菌的分离鉴定及其产贝母类生物碱分析

潘峰,苏雪梅,杨楠,蔡世美,侯凯,吴卫*   

  1. 四川农业大学农学院, 成都 611130
  • 出版日期:2018-08-08 发布日期:2018-08-08
  • 基金资助:

    高等学校博士学科点专项(20115103110009)

     

Isolation,Identification and the Analysis of Fritillaria-type Alkaloid-producing of an Endophytic Fungus from Fritillaria cirrhosa D.Don  

PAN Feng,SU Xue-mei,YANG Nan,CAI Shi-mei,HOU Kai,WU Wei*   

  1. Agronomy College,Sichuan Agricultural University,Chengdu 611130,China
  • Online:2018-08-08 Published:2018-08-08

摘要: 本文利用组织匀浆法从川贝母鳞茎中分离出一株内生真菌CBY4,采用形态学和分子生物学方法,该菌株被鉴定为Fusarium tricinctum。采用生物碱显色沉淀反应,高效液相色谱-蒸发光检测器(HPLC-ELSD)色谱法和超高效液相色谱-质谱仪(UPLC-MS)色谱法对其产贝母生物碱特征进行分析,结果显示F.tricinctum CBY4能够产生贝母类生物碱贝母辛和贝母素乙。同时利用HPLC-ELSD测定目标生物碱在发酵液中的产量,其分别为0.020 6 mg/L和 0.010 4 mg/L。本文首次报道川贝母内生真菌产生物碱特性,该研究表明内生真F.tricinctum CBY4通过液体发酵能够产生贝母辛和贝母素乙,并且具有利用生物发酵技术生产贝母辛和贝母素乙的潜在利用价值。

关键词: 川贝母, 分离鉴定, 内生真菌, 生物碱

Abstract: An endophytic fungus CBY4 was isolated from the bulbus of Fritillaria cirrhosa D.Don (FC) using tissue homogenate method.And this strain was identified as Fusarium tricinctum based on phylogenetic analysis and molecular biology analysis.In addition,the characteristics of fritillaria-type alkaloid (FTA) production were tested using alkaloid-chromogenic reaction,high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) and ultra performance liquid chromatography-mass spectrometry (UPLC-MS),followed by determining the yield of FTA using HPLC-ELSD method.And the results showed that F.tricinctum CBY4 could produce two kinds of FTA (peimisine and peiminine),and whose yields were 0.020 6 mg/L,0.010 4 mg/L in the culture filtrate,respectively.It is the first report that the characteristic of fritillaria-type alkaloid production of endophytic fungus from FC.And the work showed that endophytic fungal F.tricinctum CBY4 produced peimisine and peiminine,and has the potential value to produce peimisine and peiminine by fermentation techniques.

Key words: Fritillaria cirrhosa D.Don, isolation and identification, endophytic fungi, alkaloid

中图分类号: 

R284.1 Q946.91