凋落物真菌Neocucurbitaria salicis-albae及其代谢产物的抗细菌活性

doi:10.11924/j.issn.1000-6850.casb2023-0037

中国农学通报 ›› 2024, Vol. 40 ›› Issue (9): 145-149.doi: 10.11924/j.issn.1000-6850.casb2023-0037

所属专题：生物技术；植物保护

凋落物真菌Neocucurbitaria salicis-albae及其代谢产物的抗细菌活性

刘瑶(), 赵吴超, 杨骁, 寇雨心, 都婷婷, 陈秀青, 秦梦飞, 徐利剑()

黑龙江大学现代农业与生态环境学院，哈尔滨 150080

收稿日期:2023-01-10 修回日期:2023-05-15 出版日期:2024-03-25 发布日期:2024-03-22
通讯作者:
徐利剑，男，1981年出生，黑龙江齐齐哈尔人，教授，博士，研究方向：主要从事真菌资源与植物保护的研究。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学184信箱，Tel：0451-86609487，E-mail：xulijian@hlju.edu.cn。
作者简介:
刘瑶，男，1998年出生，黑龙江黑河人，研究生，研究方向：植物保护。通信地址：150080黑龙江省哈尔滨市南岗区学府路74号黑龙江大学现代农业与生态环境学院，Tel：0451-86609487，E-mail：1350441614@qq.com。
基金资助:
国家自然科学基金“大兴安岭林下凋落物慢速生长产孢真菌的隐秘代谢物及其抗菌活性”(31870528); 全国农业教指委2021年研究生教育研究面上课题“寒地作物绿色保护案例库建设”(2021-NYYB-64)

Antibacterial Activities of Litter Fungus Neocucurbitaria salicis-albae and Its Metabolite

LIU Yao(), ZHAO Wuchao, YANG Xiao, KOU Yuxin, DU Tingting, CHEN Xiuqing, QIN Mengfei, XU Lijian()

College of Modern Agricultural and Ecological Environment, Heilongjiang University, Harbin 150080

Received:2023-01-10 Revised:2023-05-15 Published:2024-03-25 Online:2024-03-22

摘要/Abstract

摘要：

研究旨在森林凋落物中获得具有抗菌活性的真菌资源，筛选并鉴定具有抗菌活性的真菌，纯化及鉴定其产生的抗菌化合物。以大兴安岭森林凋落物为试材，用颗粒法从中分离真菌，用基因序列比对分析测得分类学数据，用打孔药剂扩散法测得抗菌活性，采用柱层析法分离单体化合物，通过波谱分析法测得化学结构。分离得到真菌SGSF801，经多序列分析将其鉴定为Neocucurbitaria salicis-albae新株系。抗菌试验结果表明大米发酵产物对植物病原菌青枯劳尔氏菌、丁香假单胞杆菌及立枯丝核菌的抗菌活性较强。将真菌大米发酵产物分离纯化，得到一个单体化合物，通过¹H-NMR和¹³C-NMR将其鉴定为尾孢酰胺，分子式为C₁₆H₁₃NO₇。其对青枯劳尔氏菌的MIC值为31.25~62.5 μg/mL，对丁香假单胞菌的MIC值为125~250 μg/mL。本研究首次在Neocucurbitaria属真菌中分离得到尾孢酰胺，并且发现其具有抗植物病原细菌的活性，为凋落物真菌资源的进一步利用提供依据。

关键词: 真菌鉴定, 抗菌活性, 尾孢酰胺, 次生代谢产物, 大兴安岭

Abstract:

This study focuses on discovering the antimicrobial fungal resources from forest litter fungi, screening out and identifying the fungal strains with antimicrobial activities, and isolating and purifying their antimicrobial compounds. The forest litter from the Greater Hinggan Mountains was selected as test materials, then the fungi were isolated by particle separation method, the taxonomic data were measured by gene sequence comparison, the antibacterial activity was measured by well diffusion method, the monomeric compounds were isolated by column chromatography, and the chemical structure was determined by spectral analysis. Fungal strain SGSF801 was isolated and identified as a new strain of Neocucurbitaria salicis-albae by multiple sequence analysis. The results of the antimicrobial assay showed that the rice fermentation products had strong antimicrobial activities against plant pathogens Ralstonia solanacearum, Pseudomonas syringae and Rhizoctonia solani. The product of fungal rice fermentation was isolated and purified to obtain a monomeric compound, which was identified as Cercosporamide by ¹H-NMR and ¹³C-NMR, and its molecular formula was C₁₆H₁₃NO₇. Its MIC values were 31.25-62.5 μg/mL for Ralstonia solanacearum and 125-250 μg/mL for Pseudomonas syringae. In this study, Cercosporamide was isolated from fungal genus Neocucurbitaria for the first time, and it was found to have antibacterial activities against plant pathogens, which provided a basis for further utilization of litter fungi resources.

Key words: fungal identification, antibacterial activity, Cercosporamide, secondary metabolite, Greater Hinggan Mountains

刘瑶, 赵吴超, 杨骁, 寇雨心, 都婷婷, 陈秀青, 秦梦飞, 徐利剑. 凋落物真菌Neocucurbitaria salicis-albae及其代谢产物的抗细菌活性[J]. 中国农学通报, 2024, 40(9): 145-149.

LIU Yao, ZHAO Wuchao, YANG Xiao, KOU Yuxin, DU Tingting, CHEN Xiuqing, QIN Mengfei, XU Lijian. Antibacterial Activities of Litter Fungus Neocucurbitaria salicis-albae and Its Metabolite[J]. Chinese Agricultural Science Bulletin, 2024, 40(9): 145-149.

图/表 5

参考文献 22

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序列	最相近菌种	登录号	覆盖率/%	相似率/%
ITS	Neocucurbitaria salicis-albae	NR163365	100	99.10
LSU	Neocucurbitaria salicis-albae	MK442535	100	99.87
TUB2	Neocucurbitaria salicis-albae	MK442738	100	97.54

序列	最相近菌种	登录号	覆盖率/%	相似率/%
ITS	Neocucurbitaria salicis-albae	NR163365	100	99.10
LSU	Neocucurbitaria salicis-albae	MK442535	100	99.87
TUB2	Neocucurbitaria salicis-albae	MK442738	100	97.54

培养基	15℃	20℃	25℃	30℃
OA	16.00±1.63f	42.67±2.49a	41.3±1.89a	32.00±1.63bc
PDA	23.67±2.49de	29.67±1.25c	32.33±2.05bc	22.67±0.94de
YMA	20.33±2.62e	32.67±0.94bc	34.67±0.94b	30.67±0.94c
MEA	23.33±0.94de	34.67±0.94b	24.67±0.47d	24.33±0.47d

培养基	15℃	20℃	25℃	30℃
OA	16.00±1.63f	42.67±2.49a	41.3±1.89a	32.00±1.63bc
PDA	23.67±2.49de	29.67±1.25c	32.33±2.05bc	22.67±0.94de
YMA	20.33±2.62e	32.67±0.94bc	34.67±0.94b	30.67±0.94c
MEA	23.33±0.94de	34.67±0.94b	24.67±0.47d	24.33±0.47d

供试病原菌	PDA	PD	SM	大米
丁香假单胞菌	+	+	++	++
青枯劳尔氏菌	++	+	++	++
水稻黄单胞菌	—	—	—	—
玉米链格孢菌	—	—	—	—
立枯丝核菌	—	—	—	++

凋落物真菌Neocucurbitaria salicis-albae及其代谢产物的抗细菌活性

Antibacterial Activities of Litter Fungus Neocucurbitaria salicis-albae and Its Metabolite

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