生物预处理对油樟精油组成、油细胞和内生菌群的影响

doi:10.3969/j.issn.0253-2417.2021.05.008

林产化学与工业 ›› 2021, Vol. 41 ›› Issue (5): 51-57.doi: 10.3969/j.issn.0253-2417.2021.05.008

生物预处理对油樟精油组成、油细胞和内生菌群的影响

程贤¹(), 毕良武¹^,*(), 李胜男¹, 陈玉湘¹, 赵振东¹, 莫开林²

1. 中国林业科学研究院林产化学工业研究所; 生物质化学利用国家工程实验室; 国家林业和草原局林产化学工程重点实验室; 江苏省生物质能源与材料重点实验室; 江苏省林业资源高效加工利用协同创新中心, 江苏南京 210042
2. 四川省林业科学研究院, 四川成都 610081

收稿日期:2021-02-24 出版日期:2021-10-28 发布日期:2021-11-04
通讯作者: 毕良武 E-mail:chengx9013@163.com;biliangwu@126.com
作者简介:毕良武, 研究员, 博士, 博士生导师, 主要从事天然产物化学与利用研究; E-mail: biliangwu@126.com
程贤(1990-), 女, 山东济宁人, 助理研究员, 博士, 主要从事天然产物化学与利用研究; E-mail: chengx9013@163.com
基金资助:
四川省院省校合作项目(2019YFSY0002);中国林科院林产化学工业研究所研究团队建设创新工程项目(LHSXKQ2)

Effects of Biological Pretreatment on Constituents, Oil Cells Morphology and Endophytic Microbial Communities of Cinnamomum longepaniculatum

Xian CHENG¹(), Liangwu BI¹^,*(), Shengnan LI¹, Yuxiang CHEN¹, Zhendong ZHAO¹, Kailin MO²

1. Institute of Chemical Industry of Forest Products, CAF; National Engineering Lab. for Biomass Chemical Utilization; Key Lab. of Chemical Engineering of Forest Products, National Forestry and Grassland Administration; Key Lab. of Biomass Energy and Material, Jiangsu Province; Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing 210042, China
2. Sichuan Academy of Forestry Sciences, Chengdu 610081, China

Received:2021-02-24 Online:2021-10-28 Published:2021-11-04
Contact: Liangwu BI E-mail:chengx9013@163.com;biliangwu@126.com

摘要/Abstract

摘要：

分别使用加水、加入酵母菌和加入复合菌3种方式对新鲜采集的油樟叶进行预处理，提取预处理后的油樟叶的油樟精油，并利用GC-MS鉴定油樟精油的化学成分；然后利用组织透明法对油樟叶中油细胞进行形态观察，统计油细胞密度；最后通过基因测序，表征油樟叶内生菌群落结构，分析内生菌群落与生物预处理的相关性。研究结果表明：酵母菌预处理和复合菌预处理使油樟精油提取得率分别提高33.6%和49.8%，并且使主成分1，8-桉叶素的相对含量略有降低，β-蒎烯、α-松油醇和γ-松油烯等次要成分略有提高；水预处理、酵母菌预处理和复合菌预处理后油樟叶油细胞密度分别为(37±2)、(27±2)和(25±5)个/mm²，低于新鲜油樟叶的油细胞密度，(37±3个)/mm²；基于距离冗余分析(dbRDA)和相关性热图分析预处理方式对油樟叶的内生菌群落结构影响较大，其中γ-变形菌、α-变形菌、蓝细菌和酸杆菌为受影响最大的菌群，3种预处理方式对内生菌群落结构影响的程度从大到小顺序为复合菌预处理＞酵母菌预处理=水预处理；复合菌种的加入与多种核心内生菌种显著相关，尤其对浮霉菌、酸杆菌和梭状芽孢杆菌丰度的影响最为显著。

关键词: 油樟, 精油, 油细胞, 内生菌

Abstract:

The leaves of Cinnamomum longepaniculatum(Gamble) N.Chao ex H. W. Li. were pretreated by adding water, yeast and complex bacteria respectively, and the essential oil of C. longepaniculatum leaves was extracted after pretreatment. The GC-MS was used to characterize the chemical composition of essential oil. The tissue transparency method was used to observe the morphology of the oil cells after pretreatment, and calculate the density of oil cells; finally, through gene sequencing, characterize the endophytic community structure of C. longepaniculatum leaves and analyze the correlation between the endophytic bacteria community and biological pretreatment. The results showed that yeast pretreatment and complex bacteria pretreatment increased the extraction yield of essential oil by 33.6% and 49.8%, respectively, and slightly reduced the relative content of 1, 8-cineole, the minor components such as α-terpineol, β-pinene and γ-terpinene were slightly increased; the oil cell density of C.longepaniculatum leaves after pretreatment with water, yeast, and complex bacteria were (37±2), (27±2) and (25±5) per 1 mm², which were less than that of fresh leaves, (37±3) per 1 mm²; Based on distance-based redundancy analysis(dbRDA) and correlation heatmap analysis, it is found that pretreatment methods havd a greater impact on the endophytic community structure of camphor leaves. γ-Proteobacteria, α-proteobacteria, cyanobacteria and acidobacteria were the most affected species in class level. The order of the degree of influence was complex bacteria pretreatment>yeast pretreatment=water pretreatment. The addition of complex bacteria was significantly related to a variety of endophytes, especially the abundance of planktomycetes, acid bacilli and bacillus.

Key words: Cinnamomum longepaniculatum, essential oil, oil cells, endophytic microbe

中图分类号:

TQ35

程贤, 毕良武, 李胜男, 陈玉湘, 赵振东, 莫开林. 生物预处理对油樟精油组成、油细胞和内生菌群的影响[J]. 林产化学与工业, 2021, 41(5): 51-57.

Xian CHENG, Liangwu BI, Shengnan LI, Yuxiang CHEN, Zhendong ZHAO, Kailin MO. Effects of Biological Pretreatment on Constituents, Oil Cells Morphology and Endophytic Microbial Communities of Cinnamomum longepaniculatum[J]. Chemistry and Industry of Forest Products, 2021, 41(5): 51-57.

图/表 6

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图2

图3

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预处理方式 pretreatment methods	水 water	酵母菌 yeast	复合菌 complex bacteria
无blank(B)
水water (W)	1
酵母菌yeast(Y)	1	1
复合菌complex bacteria (C)	1	1	1

序号 No.	t_R/min	化合物 compound	分子式 fomula	相对峰面积relative area¹⁾/%
序号 No.	t_R/min	化合物 compound	分子式 fomula	空白 blank	水预处理 water	酵母菌预处理 yeast	复合菌预处理 complex bacteria
1	6.955	α-蒎烯α-pinenne	C₁₀H₁₆	0.31	0.42	0.29	0.50
2	7.217	β-蒎烯β-pinenne	C₁₀H₁₆	2.23	2.57	3.44	3.94
3	9.460	β-水芹烯β-phellandrene	C₁₀H₁₆	5.36	5.83	4.47	7.13
4	9.530	莰烯camphene	C₁₀H₁₆	0.05	0.15	0.44	0.28
5	12.623	邻伞花烃o-cymene	C₁₀H₁₄	0.29	0.18	0.27	0.49
6	13.415	对伞花烃p-cymene	C₁₀H₁₄	0.33	0.58	0.73	0.51
7	13.887	1, 8-桉叶素1, 8-cineole	C₁₀H₁₈O	57.25	53.89	54.58	51.56
8	19.995	罗勒烯ocimene	C₁₀H₁₆	0.05	0.37	0.36	0.78
9	31.724	侧栢烯thujene	C₁₀H₁₆	0.30	0.15	0.68	0.50
10	32.865	4-松油醇4-terpineol	C₁₀H₁₈O	4.99	3.87	2.11	2.80
11	33.945	α-松油醇α-terpineol	C₁₀H₁₈O	24.91	24.63	25.58	26.04
12	35.413	α-水芹烯α-phellandrene	C₁₀H₁₆	0.23	0	0.19	0
13	36.075	γ-松油烯γ-terpinene	C₁₀H₁₆	2.30	4.33	4.16	3.78
14	36.454	石竹烯caryophyllene	C₁₅H₂₄	0.79	1.08	0.89	0.83
15	36.659	甘香烯elixene	C₁₅H₂₄	0.37	0.71	0.22	0.43
16	36.887	大根香叶烯D germacrene D	C₁₅H₂₄	0.24	1.24	1.59	0.43

内生菌 endophyte	纲水平上内生菌相对丰度/% percent of community abundance on class level
内生菌 endophyte	空白 blank	水 water	酵母菌 yeast	复合菌 complex bacteria
γ-变形菌γ-proteobacteria	79.84	44.51	64.11	16
α-变形菌α-proteobacteria	1.32	3.13	6.04	42.52
放线菌actinobacteria	15.22	26.74	22.94	7.35
蓝细菌cyanobacteria	0	0	0	17.88
杆菌bacilli	2.64	22.74	4.12	1
酸杆菌acidobacteria	0.11	0.03	0.29	9.99
拟杆菌bacteroidia	0.51	2.16	1.6	0.88
嗜热油菌thermoleophilia	0.06	0.02	0.08	1.54
浮霉菌planctomycetes	0	0	0	1.39
其他others	0.3	0.69	0.83	1.44

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生物预处理对油樟精油组成、油细胞和内生菌群的影响

Effects of Biological Pretreatment on Constituents, Oil Cells Morphology and Endophytic Microbial Communities of Cinnamomum longepaniculatum

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