草地学报

草地学报 ›› 2025, Vol. 33 ›› Issue (1): 307-316.DOI: 10.11733/j.issn.1007-0435.2025.01.034

• 技术研发 • 上一篇    

产生物碱苦豆子内生真菌筛选及其发酵条件优化

王文凯1, 鞠明岫1, 金婧2, 刘冠兰1, 顾沛雯1   

  1. 1. 宁夏大学农学院, 宁夏 银川 750021;
    2. 银川市金凤区农业农村和水务局, 宁夏 银川 750001
  • 收稿日期:2024-03-29 修回日期:2024-05-14 发布日期:2025-01-22
  • 通讯作者: 顾沛雯,E-mail：gupeiwen2019@nxu.edu.cn
  • 作者简介:王文凯（2000-）,男,汉族,宁夏中卫人,硕士研究生,主要从事生物防治及微生物资源利用研究,E-mail：2430507742@qq.com
  • 基金资助:
    宁夏回族自治区重点研发计划项目“贺兰山东麓酿酒葡萄根部病害生物防控专用菌剂的研制与应用”（2023BCF01026）资助

Screening of Alkaloid-producing Endophytic Fungi from Sophora alopecuroides and Optimization of their Fermentation Conditions

WANG Wen-kai1, JU Ming-xiu1, JIN Jing2, LIU Guan-lan1, GU Pei-wen1   

  1. 1. Agricultral College, Ningxia University, Yinchuan, Ningxia Hui Autonomous Region 750021, China;
    2. Yinchuan Jinfeng District Agricultural Rural and Water Affairs Bureau, Yinchuan, Ningxia Hui Autonomous Region 750001, China
  • Received:2024-03-29 Revised:2024-05-14 Published:2025-01-22

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摘要： 本研究旨在筛选出能够产喹诺里西啶类生物碱的苦豆子（Sophora alopecuroides）内生真菌菌株,探究其发酵条件,提高碱产率。以前期从苦豆子健康种子中分离的50株内生真菌为材料,采用薄层色谱法、高效液相色谱法和酸性染料比色法进行产生物碱菌株的筛选,对获得的菌株进行形态学和多基因序列比对分析,确定分类地位。随后,通过单因素和响应面法探究添加L-哌啶酸、L-赖氨酸和α-酮戊二酸对产生物碱菌株碱产率的影响。结果表明,共筛选出3株高活性产槐定碱菌株DSD103,DSD201和DSD308,均属于互隔链格孢（Alternaria alternata）,其菌丝中槐定碱的产率分别为0.212,0.544和0.276 mg·g-1。碱产率最高的菌株DSD201在L-哌啶酸浓度为1.079 μmol·L-1、L-赖氨酸浓度为6.251 μmol·L-1和α-酮戊二酸浓度为0.153 μmol·L-1时,菌丝中碱产率达到最大,为0.949 mg·g-1,比对照提高了74.12%。所筛选的产生物碱菌株为通过微生物发酵生产喹诺里西啶类生物碱提供了一种新方法。

关键词: 苦豆子内生真菌, 产生物碱菌株, 筛选, 发酵条件优化, 喹诺里西啶类生物碱

Abstract: The aim of this study was to screen the endophytic fungal strains of Sophora alopecuroides that can produce Quinolizidine alkaloid, to explore the fermentation conditions and improve alkaloid production. Fifty endophytic fungal strains isolated from healthy seeds of S. alopecuroides in the previous period were used as materials. The alkaloid-producing strains were screened by Thin-Layer Chromatography, High-Performance Liquid Chromatography and Acid-dye colorimetry. The alkaloid-producing strains obtained were analyzed by morphology and multi-gene sequence comparison to determine the taxonomic status. Subsequently, the effects of the addition of precursors such as L-pipericolic acid, L-lysine and α-ketoglutaric acid on the alkaloid yields of alkaloid producing strains were explored by one-way and response surface methodology. The results showed that a total of three highly active Sophoridine-producing strains, DSD103, DSD201 and DSD308, all belonging to Alternaria alternata, were screened, and the alkaloid yields of Sophoridine in their mycelia were 0.212, 0.544 and 0.276 mg·g-1. The highest alkaloid yielding strain, DSD201, was obtained with the addition of the precursors L-pipericolic acid at a concentration of 1.079 μmol·L-1, L-lysine at a concentration of 6.251 μmol·L-1, and α-ketoglutaric acid at a concentration of 0.153 μmol·L-1. Under this conditions, the alkaloid yield in mycelium reached the maximum of 0.949 mg·g-1, which was 74.12% higher than that under the control conditions. The screened alkaloid-producing strains provide a new method for the production of Quinolizidine alkaloid by microbial fermentation.

Key words: Endophytic fungi of Sophora alopecuroides, Alkaloid-producing strains, Screening, Optimization of fermentation conditions, Quinolizidine alkaloid

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