噬菌体DCEAV-31和DCEIV-9对溶藻菌溶藻特性的影响

doi:10.13560/j.cnki.biotech.bull.1985.2022-0009

摘要/Abstract

摘要：

水华是全球关注的环境问题，而微囊藻是造成水华的主要藻类。微囊藻消长与水体理化因子和生物因子密切相关，但对感染溶藻菌的噬菌体对微囊藻的调控作用缺乏认识。本研究的目的在于理解微囊藻，溶藻菌及其噬菌体三者间的相互关系。本研究对分离自滇池的微小杆菌噬菌体DCEAV-31进行生物学特征研究，采用微囊藻-微小杆菌-噬菌体共培养体系研究三者间的相互关系。DCEAV-31属长尾噬菌体科，爆发量为28 PFU/cell，宿主域较窄，对温度、pH较敏感，对氯仿、乙醇、蛋白酶 K 和SDS 非常敏感，对Triton X-100 不敏感。微囊藻-微小杆菌-噬菌体共培养实验表明噬菌体降低了溶藻菌对微囊藻的溶藻作用，噬菌体也直接抑制了微囊藻的增殖。这些结果表明噬菌体通过裂解溶藻菌或藻际菌调控微囊藻的增殖，为我们理解微囊藻的消长提供了新的视角。

关键词: 噬菌体, 铜绿微囊藻, 溶藻菌, 相互关系, 微小杆菌

Abstract:

Water bloom is a global concerned environmental issue，and Microcystis is the main algae causing water bloom. The growth and decline of Microcystis is closely related to the physico-chemical and biotic factors in aquatic ecosystems，while the regulatory effects of bacteriophage infecting algicidal bacterium to Microcystis remains insufficient understanding. The aim of this study is to understand the interactions among Microcystis，algicidal bacterium and bacteriophage. In this study，the biological characteristics of Exiguobacterium bacteriophage DCEAV-31 isolated from Dianchi Lake were studied，and interactions among Microcystis - Exiguobacterium - bacteriophage were investigated. DCEAV-31 belonged to the Siphoviridae. Its burst size was 28 PFU/infected cell and the host range was narrow. DCEAV-31 was sensitive to temperature and pH，extremely sensitive to chloroform，ethanol，proteinase K，and SDS，but not sensitive to Triton X-100. The co-cultivation of Microcystis-Exiguobacterium-bacteriophage showed that phages reduced the algal lysis activity of algicidal bacterium to Microcystis，and bacteriophages also directly inhibited the proliferation of Microcystis. These results indicated that bacteriophages regulate the proliferation of Microcystis by lysis algicidal bacterium，which provides a new perspective for us to understand the growth and decline of Microcystis.

Key words: bacteriophage, Microcystis aeruginosa, algicidal bacterium, interaction, Exiguobacterium

张俊锋, 李孟珂, 吴志浩, 崔晓龙, 肖炜, 张仕颖. 噬菌体DCEAV-31和DCEIV-9对溶藻菌溶藻特性的影响[J]. 生物技术通报, 2022, 38(11): 250-257.

ZHANG Jun-feng, LI Meng-ke, WU Zhi-hao, CUI Xiao-long, XIAO wei, ZHANG Shi-ying. Effects of Bacteriophages DCEAV-31 and DCEIV-9 on the Algicidal Characteristics of Algicidal Bacterium Against Microcystis[J]. Biotechnology Bulletin, 2022, 38(11): 250-257.

图/表 4

表1 共培养体系试验组合

Table 1 Different assemblage setups for co-culture experi-ments

试验编号 Test No.	藻 Algae	菌 Bacterium	噬菌体 Bacteriophage
试验A	M.aeruginosa 905
试验B	M.aeruginosa 905	EA-31/EI-9
试验C	M.aeruginosa 905		DCEAV-31/DCEIV-9
试验D	M.aeruginosa 905	EA-31/EI-9	DCEAV-31/DCEIV-9

图1 噬菌体 DCEAV-31 透射电镜形态（A）和一步生长曲线（B）

Fig. 1 TEM image（A）and one-step growth curve（B）of phage DCEAV-31

图2 pH（A）和温度（B）对噬菌体 DCEAV-31 的影响

Fig. 2 Effects of pH（A）and temperature（B）on phage DCEAV-31

图3 不同试验组中各细胞和病毒颗粒的浓度 A，B：铜绿微囊藻905的细胞浓度；C，D：微小杆菌EA-31、EI-9的细胞浓度；E，F：噬菌体DCEAV-31、DCEIV-9的滴度

Fig. 3 Concentration of cells and virus particles in different experimental groups A，B：Cells concentration of M. aeruginosa 905. C，D：Cells concentration of Exiguobacterium EA-31 and EI-9. E，F：Titer of phage DCEAV-31 and DCEIV-9

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