沤竹过程中细菌菌群的结构变化和作用

doi:10.13475/j.fzxb.20211202308

纺织学报 ›› 2023, Vol. 44 ›› Issue (03): 96-103.doi: 10.13475/j.fzxb.20211202308

沤竹过程中细菌菌群的结构变化和作用

李宇¹^,², 傅佳佳¹^,²(), CAVACO-PAULO Artur¹^,², 王鸿博¹^,², 高卫东¹^,²

1.江南大学江苏省功能纺织品工程技术研究中心, 江苏无锡 214122
2.江南大学生态纺织教育部重点实验室, 江苏无锡 214122

收稿日期:2021-12-13 修回日期:2022-12-21 出版日期:2023-03-15 发布日期:2023-04-14
通讯作者: 傅佳佳(1983—),女,教授,博士。研究方向为纤维制品绿色加工技术及清洁化生产。E-mail:kathyfjj@126.com。
作者简介:李宇(1993—),女,博士生。主要研究方向为功能化纺织品。
基金资助:
国家自然科学基金面上项目(31470509);中国博士后科学基金资助项目(2019T120390);江苏省政策引导类计划项目(BZ2020010)

Structural changes and effects of bacterial community in bamboo retting

LI Yu¹^,², FU Jiajia¹^,²(), CAVACO-PAULO Artur¹^,², WANG Hongbo¹^,², GAO Weidong¹^,²

1. Jiangsu Engineering Technology Research Center of Functional Textile, Jiangnan University, Wuxi, Jiangsu 214122, China
2. Key Laboratory of Eco-Textiles (Jiangnan University), Ministry of Education, Wuxi, Jiangsu 214122, China

Received:2021-12-13 Revised:2022-12-21 Published:2023-03-15 Online:2023-04-14

摘要/Abstract

摘要：

为缩短竹材生物脱胶时间,得到能够满足后道加工的工艺纤维(竹束),并提高成品质量,探索了温水沤竹过程中细菌菌群结构的演替规律及作用机制。采用高通量测序与传统可培养技术相结合的方法,确定了沤竹过程中的优势菌;并结合水沤过程中竹块的形态变化以及竹块与竹束的化学成分差异,分析了菌群在水沤脱胶过程中的作用。结果表明:竹材的水沤脱胶过程以木质素的降解为主,以竹粉为唯一碳源筛选得到的5 株菌株,在属水平上仅占总菌群的0.70%~3.21%,优势可培养菌种为黏质沙雷氏菌(Serratia marcescens)和枯草芽孢杆菌枯草亚种(Bacillus subtilis subsp. subtilis)。筛选获得的可培养菌种中缺少起发酵作用使竹块膨胀的菌种,因此,预处理-优势菌种复配的联合工艺可以作为一种提高竹脱胶效率的手段。

关键词: 沤竹, 木质素降解, 细菌菌群, 高通量测序, 黏质沙雷氏菌, 枯草芽胞杆菌

Abstract:

Objective Bacteria produce enzymes, which can hydrolyze the ingredients from bamboo. Based on this understanding, the water-retting process is adopted to extract bamboo fiber bundles. Changes in the bacterial community structure of the water-retting liquid are investigated to reveal key functional bacteria in bamboo-retting process and to obtain high-quality bamboo fiber bundles within less time.

Method The images of the cross-section and longitudinal structure of bamboo fiber bundles were recorded by the optical microscope, and the main chemical components of bamboo fiber bundles, such as cellulose, hemicellulose, lignin, and pectin, were evaluated and compared with that of un-retted bamboo culms. High-throughput sequencing and culturable methodology were adopted to explore the dominant strains during bamboo retting. Furthermore, the changes in morphology and chemical components of bamboo were characterized to analyze the effect of bacterial community during the retting.

Results The obtained bamboo fiber bundles are composed of many single fibers which are as long as the un-retted bamboo culms. The cellulose content in bamboo fiber bundles increases by 6.56%, while the lignin content declines by 4.01% relative to un-retted bamboo culms(Tab.1). High-throughput sequencing results (Tab.2) provide 39636-57850 sequences, which are classified as 483-900 operational taxonomic units (OTUs) at the similarity level of 97%. Alpha diversity of retting liquid shows high microbial diversity in all the flora. From Fig.3 and Tab. 3, 14 phyla and 25 genera are detected from 6 samples collected at different retting time periods. At the phylum level, about 99% of the sequences belong to Proteobacteria and Firmicute. Also, Firmicute reaches the highest relative abundance of 29.25% on the 18th day of retting. Pseudomonas and Enterobacter with lignin-decomposing capability are founded as the dominant bacterial throughout the retting at the genus level. The abundances of other lignin-decomposing bacteria (Sphingomonas and Acinetobacter) are higher in the first six days. The bacteria related to the fermentation of bamboo culms belongs to Lactococcus, which is only detected on the 3rd day of retting. As the retting goes on, the abundances of Stenotrophomonas, Clostridium sensu stricto, and Clostridium IV appear to rise, resulting in the hydrolysis of hemicellulose and cellulose. The bacteria that can degrade pectin belongs to Ralstonia, taking a proportion of 10.97% on the 21st day of retting. When the bamboo powder is used as the only carbon source, 5 strains are able to multiply and account for 0.70%-3.21% of the total bacteria community at the genus level (Fig.4). Serratia marcescens and Bacillus subtilis subsp. Subtilis are regarded as key functional bacterial and these strains effectively degrade non-cellulose compounds with the cooperation of Citrobacter sp. and Klebsiella oxytoca. Macromolecule lignin in bamboo is broken into small molecules by Serratia marcescens and Citrobacter sp., and then decomposed by Bacillus subtilis subsp. Subtilis and Klebsiella oxytoca. With the degradation of lignin, Bacillus subtilis subsp. Subtilis can efficiently attack hemicellulose and pectin in the internal structure of bamboo. However, strains that can swell the bamboo culms are lacking among the culturable bacteria.

Conclusion Lignin degradation is crucial to bamboo retting. Bamboo fiber bundles in any required length can be obtained by retting with the purpose to meet the needs for subsequent processing. In all the flora, Pseudomonas and Enterobacter are the predominant bacteria and contribute to decomposing lignin, but they could not take advantage of bamboo powders as the only carbon source. Among the culturable bacteria, Serratia marcescens and Bacillus subtilis subsp. Subtilis are the primary bacteria, acting on the degradation of non-cellulose components. At the early stage of retting, bamboo swelling is not enough, so a combined treatment with pretreatment and complex use of dominant strains may efficiently improve the bamboo fiber extraction.

Key words: bamboo retting, lignin degradation, bacterial community structure, high-throughput sequencing, Serratia marcescens, Bacillus subtilis

中图分类号:

TS121.8

李宇, 傅佳佳, CAVACO-PAULO Artur, 王鸿博, 高卫东. 沤竹过程中细菌菌群的结构变化和作用[J]. 纺织学报, 2023, 44(03): 96-103.

LI Yu, FU Jiajia, CAVACO-PAULO Artur, WANG Hongbo, GAO Weidong. Structural changes and effects of bacterial community in bamboo retting[J]. Journal of Textile Research, 2023, 44(03): 96-103.

图/表 8

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图4

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竹样	纤维素/%	半纤维素/%	果胶/%	木质素/%
竹块(未经水沤)	42.05±0.61	32.94±1.26	—	18.32±0.19
竹束(水沤第24天)	48.61±0.64	33.75±1.14	0.61±0.11	14.31±0.85

样品编号	测序量	操作分类单元数量	多样性				覆盖率/%
样品编号	测序量	操作分类单元数量	香浓指数	辛普森指数	ACE	Chao1	覆盖率/%
D03	37 853	465	1.936 7	0.293 5	908.55	682.81	0.994 6
D06	49 064	900	2.063 2	0.357 2	1426.77	1295.00	0.991 9
D12	57 850	724	1.912 6	0.309 3	1575.55	1106.60	0.994 0
D18	48 518	541	2.272 2	0.273 4	886.48	833.57	0.994 9
D21	39 636	483	2.688 2	0.127 0	722.60	672.06	0.995 1
D24	41 887	504	2.503 4	0.185 0	1137.10	805.25	0.994 2

菌属名称	D03	D06	D12	D18	D21	D24
假单胞菌属	9.25	2.05	34.13	51.33	26.08	13.33
肠杆菌属	28.20	62.86	44.48	1.95	8.80	7.09
寡养单胞菌属	0.32	21.67	0.17	7.01	24.82	39.39
草螺菌属	45.75	0.06	0	2.56	14.95	10.72
乳球菌属	2.04	0	0	0	0	0.03
梭菌属IV	0	0	0.27	23.15	4.54	3.36
狭义梭菌属	0.08	0.02	5.67	2.33	2.65	0.49
未定义菌属	0.32	0.82	3.09	0.12	0.09	0.48
伯克氏菌属	0.44	0.06	0	2.61	0.67	3.89
布丘氏菌属	1.05	1.51	3.28	1.32	0.41	1.20
哈夫尼亚菌属	0.7	0.76	1.76	0.08	0.08	0
慢生根瘤菌属	0	0	0.05	0.02	0	4.14
无色杆菌属	0	0	0	0	0	4.41
不动杆菌属	3.25	1.25	0.08	0.05	0.12	0.26
沙雷氏菌属	1.52	0.73	1.86	0.12	0.06	0.54
罗尔斯顿菌属	0.03	0.01	0.03	0.04	10.97	0.51
鞘氨醇单胞菌属	4.22	1.25	0.37	0.06	0.18	0.03
苍白杆菌属	0	0	0	0.49	0	2.07
枯草杆菌属	0.01	1.38	0.99	0.8	0.69	0.03
梭菌属XlVa	0.03	0.22	0.56	2.24	2.29	0.04
欧文氏菌属	0.51	0.21	0.01	2.22	0	0.01
食酸菌属	0	0.07	0	0.08	0.03	1.54
丛毛单胞菌属	0	0	0	0.01	0	3.17
拉乌尔菌属	0.7	1.5	0.48	0.01	0	0.01

菌株编号	菌种名称	相似度/%	GenBank中的登录号	菌株所在样品
A	黏质沙雷氏菌	99	KM252937.1	D03、D06、D12、D18、D21、D24
B	枯草芽孢杆菌枯草亚种	98	GQ375227.1	D06、D12、D18、D21
C	柠檬酸杆菌	99	AB673462.1	D06
D	产酸克雷伯氏菌	100	KP420003.1	D12
E	产酸克雷伯氏菌	100	HM461887.1	D12

沤竹过程中细菌菌群的结构变化和作用

Structural changes and effects of bacterial community in bamboo retting

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