一株溶藻菌CBA02的分离、鉴定及溶藻特性研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0394

生物技术通报 ›› 2020, Vol. 36 ›› Issue (11): 55-62.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0394

一株溶藻菌CBA02的分离、鉴定及溶藻特性研究

杨冰洁¹^,²(), 向文洲¹^,³, 金雪洁¹, 陈子硕¹^,², 王灵¹^,², 吴后波¹^,³()

1.中国科学院南海海洋研究所中国科学院热带海洋生物资源与生态重点实验室,广州 510301
2.中国科学院大学,北京 100049
3.南方海洋科学与工程广东省实验室,广州 510301

收稿日期:2020-04-08 出版日期:2020-11-26 发布日期:2020-11-20
作者简介:杨冰洁,女,硕士研究生,研究方向：藻类生物技术;E-mail： ybj125660@163.com
基金资助:
国家自然科学青年科学基金项目(31602182);南方海洋科学与工程广东省实验室(广州)人才团队引进重大专项(GML2019ZD0406);中国科学院重点部署项目(ZDRW-ZS-2017-2-2);国家海洋经济创新示范城市项目(2017BH0005)

Isolation and Identification of an Algicidal Bacterium CBA02 and Its Algae-lysing Characteristics

YANG Bing-jie¹^,²(), XIANG Wen-zhou¹^,³, JIN Xue-Jie¹, CHEN Zi-shuo¹^,², WANG Ling¹^,², WU Hou-bo¹^,³()

1. Key Laboratory of Tropical Marine Bio-resources and Ecology,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301
2. University of Chinese Academy of Sciences,Beijing 100049
3. Southern Marine Science and Engineering Guangdong Laboratory,Guangzhou 510301

Received:2020-04-08 Published:2020-11-26 Online:2020-11-20

摘要/Abstract

摘要：

微藻在大规模养殖中易受溶藻微生物污染,为了鉴定一株溶藻菌CBA02的物种分类,研究该溶藻菌溶藻效应和溶藻机制。利用2216E平板法分离纯化该株细菌;采用16S rDNA基因序列对法鉴定细菌种类;以盐生杜氏藻为实验对象,探究其溶藻特性和溶藻方式。经16S rDNA鉴定,菌株CBA02初步定名为Ponticoccus sp. CBA02;CBA02对盐生杜氏藻的溶藻率随菌液发酵时间和添加量的增加而提高,随藻初始浓度的提高而降低;菌液、无菌上清液及高温灭菌处理的无菌上清液均能显著性降低盐生杜氏藻叶绿素a浓度（P<0.05）,且经pH处理之后的无菌上清液的溶藻效率随pH的升高而提高。菌株Ponticoccus sp. CBA02对盐生杜氏藻具有的强烈的溶藻作用,其溶藻活性受菌液发酵时间、添加量、藻细胞初始浓度和pH等因素的影响,CBA02是通过分泌溶藻物质间接溶藻,溶藻物质具有热稳定、适宜较高pH的特性。

关键词: 溶藻菌, 微藻养殖, 盐生杜氏藻, 溶藻特性, 溶藻物质

Abstract:

Microalgae is susceptible to be contaminated by algae-lysing microorganisms in large-scale culture. This work aims to isolate and identify an algae-lysing bacterial strain CBA02,and to study its algae-lysing effect and mechanism. The 2216E plate method was used to isolate and purify this strain,and 16S rDNA sequences to identify the strain. The algae-lysing characteristics and approaches were determined using Dunaliella salina. The strain CBA02 was identified as Ponticoccus sp. CBA02 by 16S rDNA sequences. The algicidal efficiency of CBA02 to D. salina increased with the increasing of the fermentation time and the dose of bacterial solution,while decreased with the increasing of the initial concentration of algae. The concentration of chlorophyll a of D. salina significantly decreased in the group of bacterial solution,cell-free supernatant and high-temperature treated cell-free supernatant（P<0.05）. The algicidal efficiency of the cell-free supernatant after pH treatment increased with the increasing of pH. The strain Ponticoccus sp. CBA02 presented a strong algicidal effect on D. salina,and its algicidal effect was influenced by fermentation time of bacterial solution,the dose of bacterial solution,initial concentration of algal cells and pH. CBA02 indirectly causes the lysis of D. salina by releasing water-soluble algicidal substances which have thermal stability and suitable for high pH.

Key words: algicidal bacteria, microalgae cultivation, Dunaliella salina, algae-lytic characteristics, algicidal substance

杨冰洁, 向文洲, 金雪洁, 陈子硕, 王灵, 吴后波. 一株溶藻菌CBA02的分离、鉴定及溶藻特性研究[J]. 生物技术通报, 2020, 36(11): 55-62.

YANG Bing-jie, XIANG Wen-zhou, JIN Xue-Jie, CHEN Zi-shuo, WANG Ling, WU Hou-bo. Isolation and Identification of an Algicidal Bacterium CBA02 and Its Algae-lysing Characteristics[J]. Biotechnology Bulletin, 2020, 36(11): 55-62.

图/表 7

图1 菌株CBA02的16S rDNA序列与相关菌株的系统发育树

图2 菌株Ponticoccus sp. CBA02的生长曲线

图3 Ponticoccus sp. CBA02菌液不同发酵时间对溶藻活性的影响

图4 Ponticoccus sp. CBA02菌液不同添加添加量对溶藻活性的影响

图5 盐生杜氏藻不同藻液初始浓度对溶藻活性的影响

图6 Ponticoccus sp. CBA02菌液不同处理对溶藻活性的影响标有不同小写字母者表示组间有显著性差异（P<0.05）,标有相同小写字母者表示组间无显著性差异（P>0.05）

图7 Ponticoccus sp. CBA02无菌滤液经不同pH处理后对盐生杜氏藻的溶藻效率的影响

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一株溶藻菌CBA02的分离、鉴定及溶藻特性研究

Isolation and Identification of an Algicidal Bacterium CBA02 and Its Algae-lysing Characteristics

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