两种海洋真菌混合发酵的次级代谢产物及杀虫活性研究

曹云; 侯宗敏; 董存柱; 曹凤勤; 陶敏; 余森泉; 夏玉莲; 伍显锋

doi:10.16801/j.issn.1008-7303.2021.0179

两种海洋真菌混合发酵的次级代谢产物及杀虫活性研究

doi: 10.16801/j.issn.1008-7303.2021.0179

1.
海南大学植物保护学院，热带农林生物灾害绿色防控教育部重点实验室，海口 570228

基金项目: 农业部财政项目 “海南重大入侵生物-草地贪夜蛾与红火蚁监测与防控” (1630042020020).

详细信息

作者简介:
曹云，1779913607@qq.com

通讯作者:
董存柱，czd@hainanu.edu.cn.

中图分类号: S482.3；O656.2
计量
- 文章访问数: 106
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- PDF下载量: 16
- 被引次数: 0
出版历程
- 收稿日期: 2021-06-22
- 录用日期: 2021-09-13
- 网络出版日期: 2021-12-06
- 刊出日期: 2022-04-01

Study on secondary metabolites and insecticidal activities of mixed fermentation of two marine fungi

1.
Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, College of Plant Protection, Hainan University, Haikou 570228, China

Funds: Major Invasive Species in Hainan—Monitoring and Control of Spodoptera frugiperda and Solenopsis invicta(1630042020020)

摘要

摘要: 为了研究海洋真菌哈茨木霉Trichoderma harzianum ABC19819和短柄青霉菌Penicillium Brefeldianum ABC190807混合发酵产生的次级代谢产物，利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱和薄层色谱等多种分离纯化手段对混合发酵所产生的次级代谢产物进行分离纯化，结合波谱分析技术(¹H NMR、¹³C NMR、ESI-MS)及文献对比鉴定了化合物的结构。以埃及伊蚊Aedes aegypti 3龄幼虫为研究对象，对分离鉴定得到的化合物进行活性评价。结果表明，从2株真菌混合培养的发酵产物中分离得到6个已知化合物，分别鉴定为agathic acid ( 1 )、4-羟基苯甲醛 ( 2 )、nafuredin ( 3 )、4-羟基-3,6-二甲基-2H-吡喃酮 ( 4 )、甲瓦龙酸内酯 ( 5 )和brefeldin A ( 6 )，其中化合物 1 为首次由哈茨木霉与短柄青霉菌的混合发酵物中分离得到。化合物 1 和 3 对埃及伊蚊3龄幼虫表现出明显的毒杀活性，处理72 h后的LC₅₀值分别为162 mg/L和240 mg/L。该研究对于通过混合发酵得到具有杀虫活性的化合物具有重要参考价值。
- 海洋真菌 /
- 哈茨木霉 /
- 短柄青霉菌 /
- 混合发酵 /
- 次级代谢产物 /
- 杀虫活性
Abstract: In order to study the secondary metabolites produced by the mixed fermentation of marine fungi Trichoderma harzianum ABC19819 and Penicillium brefeldianum ABC190807, silica gel, Sephadex LH-20 gel and thin-layer chromatography were used to isolate and purify the secondary metabolites produced by the mixed fermentation. The structures of the compounds were identified by ¹H NMR, ¹³C NMR, ESI-MS and compared with the data in the literatures. The activities of the compounds were evaluated using the third instar larvae of Aedes aegypti. The results showed that six known compounds were isolated from the mixed culture fermentation products of the two strains and were identified as Agathic acid ( 1 ), 4-hydroxybenzaldehyde ( 2 ), nafuredin ( 3 ), 4-hydroxy-3,6-dimethyl-2H-pyranone ( 4 ). Mevalonolactone ( 5 ) and brefeldin A ( 6 ), of which compound 1 was isolated for the first time from the mixed culture fermentation products of the two strains. Compounds 1 and 3 showed significant insecticidal activity against the third instar larvae of A. aegypti, with LC₅₀ values of 162 mg/L and 240 mg/L after 72 h of treatment. This study is of great value for obtaining compounds with insecticidal activities through mixed fermentation.
- marine fungi /
- Trichoderma harzianum /
- Penicillium brefeldianum /
- mixed fermentation /
- secondary metabolites /
- insecticidal activities

HTML全文

1 化合物1~6的化学结构

1. Structures of compounds 1-6

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表 1 500 mg/L化合物1~6对埃及伊蚊3龄幼虫的杀虫活性

Table 1. The insecticidal activities of compounds 1-6 at 500 mg/L against the third instar larvae of Aedes aegypti ($ \overline {\boldsymbol X}{\bf \pm {SD}}$, n = 3)

成分 Compounds	校正死亡率 ($ \overline { X}\pm { \rm {SD}}$, n = 3) Adjusted mortality rate/%
成分 Compounds	12 h	24 h	36 h	72 h
1	73.33±7.64 b	83.33±2.89 b	86.67±2.89 b	100.00±0.00 a
2	—	—	—	6.67±3.33 c
3	58.33±2.89 c	70.00±5.00 c	78.33±5.77 c	81.66±2.89 b
4	—	—	—	—
5	—	—	—	8.33±2.89 c
6	—	—	—	—
CK	—	—	—	—
鱼藤酮 rotenone	100.00±0.00 a	100.00±0.00 a	100.00±0.00 a	100.00±0.00 a
注：同列数据后不同小写字母表示差异显著 (P<0.05，DMRT法) ；“—” 表示无毒杀作用。Note: Data followed different letters within colum indicate significant differences at 0.05 level. "—" indicate no larvicidal activity.

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表 2 72 h化合物1和3对埃及伊蚊3龄幼虫的LC₅₀值

Table 2. LC₅₀ value of compound 1 and 3 after 72 h against the third instar larvae of Aedes aegypti

成分 Compound	毒力回归曲线 Regression equation of toxicity	相关系数 Correlation coefficient	LC₅₀/(mg/L)	95% 置信区间 95% confidence interval /(mg/L)
1	y = 2.1967x + 0.1413	0.9366	162	137~192
3	y = 2.2440x − 0.3418	0.9700	240	205~280
鱼藤酮 rotenone	y = 1.4031x + 3.4167	0.9894	13.4	5.72~17.5

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