放线菌4-1的鉴定及其发酵液抑菌作用和稳定性探究

doi:10.11924/j.issn.1000-6850.casb2023-0135

中国农学通报 ›› 2024, Vol. 40 ›› Issue (9): 117-123.doi: 10.11924/j.issn.1000-6850.casb2023-0135

所属专题：生物技术

放线菌4-1的鉴定及其发酵液抑菌作用和稳定性探究

郭宇豪¹(), 刘绍波¹, 葛菁萍¹^,²(), 平文祥¹^,²()

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，黑龙江省普通高校分子生物学重点实验室，哈尔滨 150080
² 河北环境工程学院，河北省农业生态安全重点实验室，河北秦皇岛 066102

收稿日期:2023-03-23 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:
葛菁萍，女，1972年出生，黑龙江齐齐哈尔人，教授，博士，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号黑龙江大学224信箱，Tel：0451-86609016，E-mail：gejingping@126.com。
平文祥，男，1959年出生，黑龙江哈尔滨人，教授，博士，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号黑龙江大学224信箱，Tel：0451-86609016，E-mail：wenxiangp@aliyun.com。
作者简介:
郭宇豪，男，2000年出生，黑龙江牡丹江人，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号黑龙江大学生命科学学院，Tel：0451-86609016，E-mail：guoyuhao0628@163.com。
基金资助:
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”(ZD2020C008); 黑龙江省农业废弃物综合利用体系岗位专家项目“黑龙江省现代农业产业技术协同创新推广体系作物秸秆低温降解菌株的研制”; 黑龙江省生态环境厅2022年度生态环境保护科研项目“人工重组细菌菌群对黑土磷障碍的解控机制”(HST2022TR004); 龙江省高校基本科研业务费“群体感应介导下重组根圈细菌菌群对土壤磷养分循环的调控机制”(2023-KYYWF-1448)

Identification of Actinomyces 4-1 and Study on Bacteriostasis and Stability of Its Fermentation Broth

GUO Yuhao¹(), LIU Shaobo¹, GE Jingping¹^,²(), PING Wenxiang¹^,²()

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 Molecular Biology, College of Heilongjiang Province/School of Life Sciences, Heilongjiang University, Harbin 150080
² Hebei University of Environmental Engineering/Hebei Key Laboratory of Agroecological Safety, Qinhuangdao, Hebei 066102

Received:2023-03-23 Revised:2023-05-15 Published:2024-03-25 Online:2024-03-22

摘要/Abstract

摘要：

旨在挖掘盐碱地菌种资源并对其抑菌功能进行检测。以自行分离的放线菌4-1为候选菌株，利用形态学、生理生化以及分子生物学手段对其种属地位进行综合鉴定；采用琼脂扩散法和菌丝生长速率抑制法对其发酵液抑菌活性和稳定性进行评价。结果表明，菌株4-1与紫赤链霉菌(Streptomyces puniceus)最为相似，同源性高达99.93%，建立的系统发育树中二者亲缘关系最为相近；菌株4-1发酵液的抑菌谱比较广泛，对病原性细菌（6株）和病原性真菌（5株）都具有一定的抑制效果；酸碱、热、紫外线对发酵液稳定性影响较小，但当温度高于90℃或pH < 4时，发酵液抑菌活性下降。该菌种资源的获得及功能评价，为进一步扩大根际促生细菌种类及应用奠定了菌种基础。

关键词: 放线菌, 紫赤链霉菌, 菌株鉴定, 抑菌作用, 稳定性

Abstract:

In order to explore the bacteria resources of saline-alkali soil and test their antibacterial function, actinomycete 4-1 was isolated and used as the original strain. Combined with morphological characteristics, physiological and biochemical indicators, as well as molecular biology techniques, the species status of 4-1 was comprehensively identified, while the antibacterial activity and stability of its fermentation broth were evaluated using agar diffusion method and mycelial growth rate inhibition method. The results showed that strain 4-1 showed the highest similarity to Streptomyces puniceus, with a homology of 99.93%, and the constructed phylogenetic tree showed that they were most closely related. The fermentation broth of strain 4-1 had a wide range of antibacterial spectrum and showed certain inhibitory effects on pathogenic bacteria (6 strains) and pathogenic fungi (5 strains). The stability of the fermentation broth was minimally affected by acidity, alkalinity, heat, or ultraviolet radiation. However, the antibacterial activity of the fermentation broth decreased when the temperature exceeded 90℃ or the pH was below 4. The availability and functional evaluation of this strain laid a foundation for further expanding rhizosphere growth-promoting bacteria resources and applications.

Key words: actinomyces, Streptomyces puniceus, strain identification, bacteriostasis, stability

郭宇豪, 刘绍波, 葛菁萍, 平文祥. 放线菌4-1的鉴定及其发酵液抑菌作用和稳定性探究[J]. 中国农学通报, 2024, 40(9): 117-123.

GUO Yuhao, LIU Shaobo, GE Jingping, PING Wenxiang. Identification of Actinomyces 4-1 and Study on Bacteriostasis and Stability of Its Fermentation Broth[J]. Chinese Agricultural Science Bulletin, 2024, 40(9): 117-123.

图/表 7

参考文献 30

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供试细菌	革兰氏染色	抑菌圈直径/mm
枯草芽孢杆菌Bacillus subtilis	阳性	15.5 ± 0.17 (＋＋＋)
金黄色葡萄球菌Staphylococcus aureus	阳性	16.5 ± 0.18 (＋＋＋)
蜡样芽胞杆菌Bacillus cereus	阳性	12 ± 0.45 (＋＋＋)
短小芽孢杆菌Bacillus pumilus	阳性	16 ± 0.20 (＋＋)
铜绿假单胞菌Pseudomonas aeruginosa	阴性	13 ± 0.34 (＋＋)
变形杆菌Proteus vulgaris	阴性	12 ± 0.50 (＋＋)
大肠杆菌Escherichia coil	阴性	0 (－)
肺炎克雷伯氏菌Klebsiella pneumoniae	阴性	0 (－)
阴沟肠杆菌Enterobacter cloacae	阴性	0 (－)

供试细菌	革兰氏染色	抑菌圈直径/mm
枯草芽孢杆菌Bacillus subtilis	阳性	15.5 ± 0.17 (＋＋＋)
金黄色葡萄球菌Staphylococcus aureus	阳性	16.5 ± 0.18 (＋＋＋)
蜡样芽胞杆菌Bacillus cereus	阳性	12 ± 0.45 (＋＋＋)
短小芽孢杆菌Bacillus pumilus	阳性	16 ± 0.20 (＋＋)
铜绿假单胞菌Pseudomonas aeruginosa	阴性	13 ± 0.34 (＋＋)
变形杆菌Proteus vulgaris	阴性	12 ± 0.50 (＋＋)
大肠杆菌Escherichia coil	阴性	0 (－)
肺炎克雷伯氏菌Klebsiella pneumoniae	阴性	0 (－)
阴沟肠杆菌Enterobacter cloacae	阴性	0 (－)

供试病原真菌	对照菌落直径/mm	处理菌落直径/mm	抑制率/%
小麦赤霉菌Gibberella zeae	85.0 ± 0.30	61.0 ± 0.17	29.41
禾谷镰刀菌Fusarium graminearum	60.0 ± 0.17	18.0 ± 0.17	70.00
辣椒根腐菌Phytophthora pepper	42.30 ± 0.20	37.65 ± 0.17	11.00
串珠镰刀菌Fusarium moniliforme sheld	74.50 ± 0.10	51.00 ± 0.17	31.54
瓜果腐霉菌Pythium melonii	84.50 ± 0.20	41.5 ± 0.17	50.90

供试病原真菌	对照菌落直径/mm	处理菌落直径/mm	抑制率/%
小麦赤霉菌Gibberella zeae	85.0 ± 0.30	61.0 ± 0.17	29.41
禾谷镰刀菌Fusarium graminearum	60.0 ± 0.17	18.0 ± 0.17	70.00
辣椒根腐菌Phytophthora pepper	42.30 ± 0.20	37.65 ± 0.17	11.00
串珠镰刀菌Fusarium moniliforme sheld	74.50 ± 0.10	51.00 ± 0.17	31.54
瓜果腐霉菌Pythium melonii	84.50 ± 0.20	41.5 ± 0.17	50.90

放线菌4-1的鉴定及其发酵液抑菌作用和稳定性探究

Identification of Actinomyces 4-1 and Study on Bacteriostasis and Stability of Its Fermentation Broth

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