副干酪乳杆菌与芽孢杆菌属共培养种间关系对产细菌素的影响

doi:10.11924/j.issn.1000-6850.casb2020-0724

中国农学通报 ›› 2021, Vol. 37 ›› Issue (24): 124-132.doi: 10.11924/j.issn.1000-6850.casb2020-0724

副干酪乳杆菌与芽孢杆菌属共培养种间关系对产细菌素的影响

姜雪雍¹^,²(), 岳元春¹, 孙养存¹^,², 高冬妮¹^,², 平文祥¹^,², 葛菁萍¹^,²()

¹黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 150080
²微生物黑龙江省高校重点实验室/黑龙江大学生命科学学院,哈尔滨 150500

收稿日期:2020-11-30 修回日期:2020-12-28 出版日期:2021-08-25 发布日期:2021-08-27
通讯作者: 葛菁萍
作者简介:姜雪雍,女,1996年出生,黑龙江佳木斯人,硕士,研究方向：微生物资源挖掘与利用。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号224信箱黑龙江大学生命科学学院,Tel：0451-86609016,E-mail： jiangxueyong7@126.com。
基金资助:
国家自然科学基金面上项目“乙酸溢流代谢与群体感应的协同效应对副干酪乳杆菌HD1.7种群稳定性影响机制的探究”(32071519);黑龙江省自然科学基金重点项目“乙酸代谢与副干酪乳杆菌群体感应互作探究及对细菌素产生的影响”(ZD 2020C 008);“黑龙江大学研究生创新科研项目”(YJSCX2020-201HLJU)

Interspecific Relationship of Lactobacillus paracasei Co-cultured with Bacillus sp. Affects Bacteriocin Production

Jiang Xueyong¹^,²(), Yue Yuanchun¹, Sun Yangcun¹^,², Gao Dongni¹^,², Ping Wenxiang¹^,², Ge Jingping¹^,²()

¹Agricultural Microorganisms Technology Education Engineering Research Center, Harbin 150080
²Key Laboratory of Microbiology of Heilongjiang Province, Life Science College, Heilongjiang University, Harbin 150500

Received:2020-11-30 Revised:2020-12-28 Online:2021-08-25 Published:2021-08-27
Contact: Ge Jingping

摘要/Abstract

摘要：

为使芽孢杆菌属与副干酪乳杆菌(Lactobacillus paracasei)通过种间信息交流形成稳定的小生态环境,加大芽孢杆菌属的初始接种量,来探究L. paracasei HD1.7与其共培养的生态学关系。结果表明,将枯草芽孢杆菌(Bacillus subtilis)与L. paracasei HD1.7以5%：1%的初始接种比例共培养,L. paracasei HD1.7所产细菌素为其单培养的1.21倍,群体感应相关基因luxS,prcK和prcR分别上调表达3.14、4.98和5.41倍。同时,B. subtilis形成更多的芽孢以回避细菌素的攻击,芽孢形成相关基因spo0A,sigE,sigF和sigG分别上调2.37、2.71、3.15和2.56倍。此时,二者形成了一种协同合作的生态关系。冗余分析结果表明,共培养体系中与细菌素产生关系最为密切的是芽孢数量、培养时间以及B. subtilis活菌数。L. paracasei HD1.7与芽孢杆菌属之间存在不同的生态关系,有与之共培养后对L. paracasei HD1.7偏利的芽孢杆菌,也存在对其偏害的芽孢杆菌。

关键词: 副干酪乳杆菌, 芽孢杆菌属, 共培养, 细菌素, 种间关系, 次生代谢产物, 生物胁迫

Abstract:

To form a stable ecological environment through interspecific information exchange between Lactobacillus paracasei HD1.7 and Bacillus sp., the initial inoculation amount of Bacillus sp. was increased to study the ecological relationship of Lactobacillus paracasei co-cultured with Bacillus sp.. When Bacillus subtilis was co-cultured with L. paracasei HD1.7 at the initial inoculation ratio of 5%:1%, the bacteriocin produced by L. paracasei HD1.7 was 1.21 times that of monoculture. L. paracasei HD1.7 quorum sensing-related genes luxS, prcK, and prcR were upregulated (up to 3.14, 4.98 and 5.41 folds, respectively) while B. subtilis formed more spores to avoid the attack of bacteriocins. Expression levels of spo0A, sigE, sigF, and sigG, genes related to B. subtilis endospore formation, were correspondingly upregulated (up to 2.37, 2.71, 3.15 and 2.56 folds, respectively). At this time, a cooperative ecological relationship was formed between L. paracasei HD1.7 and Bacillus sp.. The redundancy analysis revealed that the number of spores, incubation time and B. subtilis viable counts were the most closely related to the production of bacteriocin. Different ecological relationships were found between L. paracasei HD1.7 and Bacillus sp., and some Bacillus are beneficial to L. paracasei HD1.7 and some are harmful to it.

Key words: Lactobacillus paracasei, Bacillus sp., co-culture, bacteriocin, interspecific relationship, secondary metabolite, biotic stress

中图分类号:

S182

姜雪雍, 岳元春, 孙养存, 高冬妮, 平文祥, 葛菁萍. 副干酪乳杆菌与芽孢杆菌属共培养种间关系对产细菌素的影响[J]. 中国农学通报, 2021, 37(24): 124-132.

Jiang Xueyong, Yue Yuanchun, Sun Yangcun, Gao Dongni, Ping Wenxiang, Ge Jingping. Interspecific Relationship of Lactobacillus paracasei Co-cultured with Bacillus sp. Affects Bacteriocin Production[J]. Chinese Agricultural Science Bulletin, 2021, 37(24): 124-132.

图/表 9

图1. 不同接种比例单培养以及共培养体系中pH的变化(A)和B. subtilis活菌数(B)

图2. 不同接种比例单培养以及共培养体系中L. paracasei HD1.7活菌数(A)及其抑菌活性(B) *：P<0.05,**：P<0.01

比例/时间	0h	4h	8h	12h	24h	36h	48h	60h
3%:1%	1.750	0.875	2.000	5.923	8.250	3.682	1.724	2.563
5%:1%	0.556	0.444	0.900	8.429	6.444	5.947	5.957	3.829
5%:2%	0.556	0.556	0.600	0.786	4.389	4.880	4.044	2.714

表1. 不同接种比例共培养与单培养中B. subtilis产芽孢数之比

图3. L. paracasei HD1.7与B. subtilis共培养体系中物种与环境变量相关关系的冗余分析(RDA)

图4. 不同比例下L. paracasei HD1.7群体感应相关基因luxS、prcK和prcR(A)以及B. subtilis芽孢形成基因spo0A、sigE、sigF和sigG(B)相对表达量的差异

图5. 6种芽孢杆菌5%接种量单培养(A)及与L. paracasei HD1.7以5%:1%的比例共培养(B)的发酵液pH变化情况

图6. 6种芽孢杆菌5%接种量单培养(A)及与L. paracasei HD1.7以5%:1%的比例共培养(B)菌体生长变化情况

菌株/时间	0h	4h	8h	12h	24h	36h	48h	60h
B. licheniformis	0	0.13	1.80	4.00	3.57	3.02	2.93	4.00
B. megaterium	1.00	0.13	0.80	1.37	3.65	2.23	1.76	1.88
B. pumilus	0.18	0.10	0.32	2.70	5.50	6.00	3.46	4.88
B. cereus	0.76	0.29	0.42	0.40	0.40	1.05	1.01	1.02
B. laterosporus	0.45	0.15	0.43	1.55	1.30	7.50	1.55	1.24
B. thuringiensis	0.75	0.33	1.20	1.50	2.33	1.48	1.26	1.14

表2. 芽孢杆菌属产芽孢共培养与单培养之比

图7. L. paracasei HD1.7与6种芽孢杆菌以5:1的比例共培养L. paracasei HD1.7菌体生长情况以及产细菌素情况 *：P<0.05,**：P<0.01

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副干酪乳杆菌与芽孢杆菌属共培养种间关系对产细菌素的影响

Interspecific Relationship of Lactobacillus paracasei Co-cultured with Bacillus sp. Affects Bacteriocin Production

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