工业大麻内生真菌HDM07挥发性成分分析

doi:10.11924/j.issn.1000-6850.casb2024-0055

中国农学通报 ›› 2024, Vol. 40 ›› Issue (30): 103-113.doi: 10.11924/j.issn.1000-6850.casb2024-0055

工业大麻内生真菌HDM07挥发性成分分析

邢翔宇¹(), 丛添玥¹, 关昕¹, 祁可香¹, 李宛儒¹, 张赫¹, 尤梦瑶¹, 吴桐²(), 郑春英¹()

¹ 黑龙江大学，生命科学学院，农业微生物技术教育部工程研究中心，黑龙江省寒区植物基因与生物发酵重点实验室，黑龙江省普通高校微生物重点实验室，哈尔滨 150080
² 黑龙江大学，生命科学学院，农业微生物技术教育部工程研究中心，黑龙江省寒区植物基因与生物发酵重点实验室，黑龙江省普通高校分子生物学重点实验室，哈尔滨 150080

收稿日期:2024-01-10 修回日期:2024-04-15 出版日期:2024-10-25 发布日期:2024-10-23
通讯作者:
郑春英，女，1968年出生，黑龙江哈尔滨人，教授，博士，主要从事食品和药物生物活性挖掘及研发。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院，Tel：0451-86608586，E-mail：zhengchunying68@163.com；
吴桐，女，1979年出生，黑龙江哈尔滨人，讲师，学士，主要从事寒区植物基因与生物发酵研究。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院，Tel：0451-86608586，E-mail：523791254@qq.com。
作者简介:
邢翔宇，男，2002年出生，黑龙江大庆人，硕士研究生，研究方向：食品和药物生物活性挖掘及研发。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院，Tel：0451-886608586，E-mail：930504521@qq.com。
基金资助:
黑龙江省种业创新发展资金项目“寒地龙药重点中药材种子种苗DNA指纹图谱库、特征库及质量控制体系研发与推广”(20230202); 黑龙江省现代农业产业技术协同创新体系项目“麻类食品与药物研发协同创新项目”(YYM19STX-1); 黑龙江大学横向项目“工业大麻内生真菌及其次生代谢物提取、分离、鉴定”(21192); 黑龙江大学横向项目“一株产大麻酚的大麻内生真菌及其应用”(23114); 黑龙江大学横向项目“一种利用甘草内生真菌发酵工业大麻的方法”(23113)

Analysis of Volatile Components of Endophytic Fungus HDM07 in Cannabis sativa L.

XING Xiangyu¹(), CONG Tianyue¹, GUAN Xin¹, QI Kexiang¹, LI Wanru¹, ZHANG He¹, YOU Mengyao¹, WU Tong²(), ZHENG Chunying¹()

¹ Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080
² Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education &Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Molecular Biology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2024-01-10 Revised:2024-04-15 Published:2024-10-25 Online:2024-10-23

摘要/Abstract

摘要：

旨在开发利用工业大麻内生真菌HDM07挥发性成分，探索工业大麻内生真菌HDM07次生代谢产物生物合成途径。以工业大麻内生真菌HDM07为研究对象，采用GC-MS法和检索质谱数据库分析鉴定内生真菌HDM07发酵液和菌丝体中的挥发性成分，采用形态学观察法和18S rDNA序列分析法鉴定菌种。结果表明，经GC-MS法和质谱数据库检索分析，从内生真菌HDM07发酵液中鉴定出11种挥发性成分，在内生真菌HDM07菌丝体中鉴定出14种挥发性成分，工业大麻叶中鉴定了12种成分，空白对照品溶液（PDB培养基）中鉴定了7种成分；经鉴定内生真菌HDM07为链格孢菌(Alternaria alternate)。上述结果表明，HDM07发酵液挥发性成分与菌丝体中挥发性成分两者的合成途径既有联系又有区别。内生真菌HDM07的发酵液及菌丝体中均含有与工业大麻相同或相似的挥发性成分，也含有其特有的挥发性成分，说明该菌株与宿主植物工业大麻在合成挥发性成分方面具有一定的相关性。

关键词: 工业大麻, 工业大麻内生真菌HDM07, 18S rDNA序列分析, 气相色谱质谱法, 挥发性成分

Abstract:

This study aims to exploit volatile components of endophytic fungus HDM07 isolated from Cannabis sativa L., and to explore the biosynthetic pathway of secondary metabolites from HDM07. The research took the endophytic fungus HDM07 obtained from Cannabis sativa L. as the subject. GC-MS and retrieval mass spectrometry database were employed to identify the volatile components in both the fermentation broth and mycelium of HDM07. Morphological observation and 18S rDNA sequence analysis were conducted for strain identification. Results indicated that the method of GC-MS and mass spectrometry analysis, along with database retrieval from endophytic fungi HDM07, determined 11 volatile components in the fermented liquid. Furthermore, fungus HDM07 revealed 14 volatile constituents in mycelium and Cannabis sativa L. leaf identified 12 elements. In a blank reference substance solution (PDB medium), appraisal revealed 7 ingredients. The endophytic fungus HDM07 was identified as Alternaria alternate. These findings suggest that the synthesis pathways of volatile components in HDM07 fermentation broth and mycelia are both related yet distinct. Both the fermentation broth and mycelium of endophytic fungus HDM07 contained similar or identical volatile components to those found in Cannabis sativa L., and also contained unique ones, indicating a certain correlation between the strain and its host plant's synthesis of volatile components.

Key words: Cannabis sativa L., endophytic fungus HDM07 from Cannabis sativa L., 18S rDNA sequence analysis, GC-MS, volatile component

邢翔宇, 丛添玥, 关昕, 祁可香, 李宛儒, 张赫, 尤梦瑶, 吴桐, 郑春英. 工业大麻内生真菌HDM07挥发性成分分析[J]. 中国农学通报, 2024, 40(30): 103-113.

XING Xiangyu, CONG Tianyue, GUAN Xin, QI Kexiang, LI Wanru, ZHANG He, YOU Mengyao, WU Tong, ZHENG Chunying. Analysis of Volatile Components of Endophytic Fungus HDM07 in Cannabis sativa L.[J]. Chinese Agricultural Science Bulletin, 2024, 40(30): 103-113.

图/表 13

参考文献 33

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仪器	型号规格	生产厂家
电子精密天平	PL303	梅特勒-托利多仪器有限公司
超净工作台	SZX	南通科学仪器有限公司
空气浴振荡器	HZQ-C	哈尔滨市东联电子技术开发有限公司
旋转蒸发仪	R-201	上海申胜生物技术有限公司
超声波清洗器	KQ-100DE	昆山市超声仪器有限公司
紫外可见波长检测器	FL2000	浙江温岭福立分析仪器有限公司
高效液相色谱仪	FL2000	浙江温岭福立分析仪器有限公司

仪器	型号规格	生产厂家
电子精密天平	PL303	梅特勒-托利多仪器有限公司
超净工作台	SZX	南通科学仪器有限公司
空气浴振荡器	HZQ-C	哈尔滨市东联电子技术开发有限公司
旋转蒸发仪	R-201	上海申胜生物技术有限公司
超声波清洗器	KQ-100DE	昆山市超声仪器有限公司
紫外可见波长检测器	FL2000	浙江温岭福立分析仪器有限公司
高效液相色谱仪	FL2000	浙江温岭福立分析仪器有限公司

序号	分子式	化合物名称	相对含量/%	保留时间/min
1	C₄H₆S	硫化二乙烯	0.81	12.34
2	C₈H₁₀O₂	对羟基苯乙醇	1.40	27.99
3	C₂₄H₃₉FO₂	邻氟苯甲酸十七烷基酯	0.63	53.51
4	C₁₀H₁₄O₂	3-异丙氧基-5-甲基苯酚	0.16	80.17
5	C₂₆H₄₂O₄	丁基十四烷酞酸酯	0.04	92.28
6	C₁₆H₂₂O₄	邻苯二甲酸二丁酯	3.50	104.20
7	C₂₇H₄₂O₄	2-(3-乙酰氧基-4,4,14-三甲基雄甾-8-烯-17-基)-丙酸	0.04	115.39
8	C₂₂H₄₂O₄	己二酸二辛酯	6.43	140.00
9	C₂₂H₄₃NO	芥酸酰胺	0.69	181.47
10	C₂₈H₄₀O	麦角甾-4,6,8(14),22-四烯-3-酮	0.08	212.19
11	C₃₅H₆₂O₃	3,5-二(1,1-二甲基乙基)-4-羟基-苯丙酸十八烷基酯	0.51	228.79

序号	分子式	化合物名称	相对含量/%	保留时间/min
1	C₄H₆S	硫化二乙烯	0.81	12.34
2	C₈H₁₀O₂	对羟基苯乙醇	1.40	27.99
3	C₂₄H₃₉FO₂	邻氟苯甲酸十七烷基酯	0.63	53.51
4	C₁₀H₁₄O₂	3-异丙氧基-5-甲基苯酚	0.16	80.17
5	C₂₆H₄₂O₄	丁基十四烷酞酸酯	0.04	92.28
6	C₁₆H₂₂O₄	邻苯二甲酸二丁酯	3.50	104.20
7	C₂₇H₄₂O₄	2-(3-乙酰氧基-4,4,14-三甲基雄甾-8-烯-17-基)-丙酸	0.04	115.39
8	C₂₂H₄₂O₄	己二酸二辛酯	6.43	140.00
9	C₂₂H₄₃NO	芥酸酰胺	0.69	181.47
10	C₂₈H₄₀O	麦角甾-4,6,8(14),22-四烯-3-酮	0.08	212.19
11	C₃₅H₆₂O₃	3,5-二(1,1-二甲基乙基)-4-羟基-苯丙酸十八烷基酯	0.51	228.79

序号
1	C₁₂H₃₆O₆Si₆	十二甲基环六硅氧烷	1.32	17.13
2	C₁₄H₄₂O₇Si₇	十四甲基环七硅氧烷	0.78	32.52
3	C₂₄H₃₉FO₂	邻氟苯甲酸十七烷基酯	1.26	53.51
4	C₁₈H₃₆O	十八烷醛	0.29	69.13
5	C₁₆H₂₂O₄	邻苯二甲酸二丁酯	3.69	104.14
6	C₁₆H₃₂O₂	棕榈酸	1.13	111.93
7	C₁₈H₃₅NO	油酸酰胺	0.55	137.29
8	C₂₀H₆₀O₁₀Si₁₀	二十甲基环十硅氧烷	0.50	150.60
9	C₂₁H₃₈O₄	顺9,12-十八碳二烯酸-2,3-二羟基丙基酯	0.43	171.02
10	C₃₀H₅₀	2,6,10,15,19,23-六甲基-2,6,10,14,18,22-二十四碳六烯	1.37	186.14
11	C₂₈H₄₂O	脱氢麦角甾醇	1.81	193.35
12	C₂₈H₄₄O	麦角甾醇	1.45	205.00
13	C₂₈H₄₀O	麦角甾-4,6,8(14),22-四烯-3-酮	0.40	212.19
14	C₃₅H₆₂O₃	3,5-二(1,1-二甲基乙基)-4-羟基-苯丙酸十八烷基酯	0.10	228.79

工业大麻内生真菌HDM07挥发性成分分析

Analysis of Volatile Components of Endophytic Fungus HDM07 in Cannabis sativa L.

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序号	分子式	化合物名称	相对含量/%	保留时间/min
1	C₅H₁₀O₄	1,2,3-丙三醇-乙酸酯	0.63	8.94
2	C₁₅H₂₄	石竹烯	1.66	25.84
3	C₂₄H₃₉FO₂	邻氟苯甲酸十七烷基酯	0.14	53.51
4	C₆H₁₂O₆	d-甘露糖	0.06	66.75
5	C₁₂H₂₀O	5,5,8a-三甲基-3,5,6,7,8,8a-六氢-2H-色烯	0.22	80.60
6	C₁₈H₃₆O	6,10,14-三甲基-2-十五烷酮	0.35	91.30
7	C₁₆H₂₂O₄	邻苯二甲酸二丁酯	7.07	104.89
8	C₂₀H₄₀O	叶绿醇	1.87	116.88
9	C₂₁H₃₀O₂	大麻二酚	16.26	142.76
10	C₃₀H₅₀	2,6,10,15,19,23-六甲基-2,6,10,14,18,22-二十四碳六烯	0.77	185.87
11	C₃₄H₇₀	三十四烷	2.78	194.92
12	C₃₀H₅₀O	α-香树脂醇	1.06	209.19

序号		分子式	化合物名称			相对含量/%		保留时间/min
1		C₈H₈O	苯乙醛			0.22		7.9
2		C₆H₆N₂O	烟酰胺			0.10		23.53
3		C₁₄H₂₂O	2,6-二叔丁基苯酚			0.03		36.39
4		C₁₆H₂₂O₄	邻苯二甲酸二丁酯			1.52		103.58
5		C₂₃H₃₂O₂	2,2'-亚甲基二[6-(1,1-二甲基乙基)-4-甲基苯酚			0.06		139.09
6	C₂₄H₃₈O₄			邻苯二甲酸二(2-乙基己)酯	0.12		152.63
7	C₂₂H₄₃NO			芥酸酰胺	0.22		180.84

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