Assessment of different mcy genes for detecting the toxic to non-toxic Microcystis ratio in the field by multiplex qPCR  

作　　者：ZUO Jun CHEN Liting SHAN Kun HU Lili SONG Lirong GAN Nanqin 左俊;陈莉婷;闪锟;胡丽丽;宋立荣;甘南琴(State Key Laboratory of Freshwater Ecology and Biotechnology,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China;University of Chinese Academy of Sciences,Beijing 100049,China;Institute of Electronic Information Technology,Chongqing Institute of Green and Intelligent Technology,Chinese Academy of Sciences,Chongqing 400714,China)

机构地区：[1]State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China [2]University of Chinese Academy of Sciences, Beijing 100049, China [3]Institute of Electronic Information Technology, Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences, Chongqing 400714, China

出　　处：《Journal of Oceanology and Limnology》2018年第4期1132-1144,共13页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(Nos.31370418,41561144008);the Jiangxi Water Science and Technology Fund(No.KT201602);the State Key Laboratory of Freshwater Ecology and Biotechnology(No.2016FBZ07)

摘　　要：Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis and non-microcystin-producing Microcystis are key to predicting and treating Microcystis blooms. Multiplex qPCR is a useful tool to assess such issues. In this study, we developed multiplex qPCR methods with newly-designed probes and primers for the microcystin-synthesis related genes mcyA and mcyE. We used seven toxic Microcystis strains and four non-toxic Microcystis strains to compare the differences in the ratios of toxic and non-toxic Microcystis in mixed cultures, which were calculated using abundances of the genes mcyA, mcyB, mcyD, mcy E and phycocyanin( PC). We also compared traditional cell counting and multiplex qPCR. Hierarchical clustering and principal component analysis indicated that mcyD was the most suitable mcy gene for quantification in laboratory experiments. mcyB abundances were always higher; we suggest that the amount of toxic Microcystis measured using mcyB might overestimate the actual percentages.Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis and non-microcystin-producing Microcystis are key to predicting and treating Microcystis blooms. Multiplex qPCR is a useful tool to assess such issues. In this study, we developed multiplex qPCR methods with newly-designed probes and primers for the microcystin-synthesis related genes mcyA and mcyE. We used seven toxic Microcystis strains and four non-toxic Microcystis strains to compare the differences in the ratios of toxic and non-toxic Microcystis in mixed cultures, which were calculated using abundances of the genes mcyA, mcyB, mcyD, mcy E and phycocyanin（ PC）. We also compared traditional cell counting and multiplex qPCR. Hierarchical clustering and principal component analysis indicated that mcyD was the most suitable mcy gene for quantification in laboratory experiments. mcyB abundances were always higher; we suggest that the amount of toxic Microcystis measured using mcyB might overestimate the actual percentages.

关 键 词：Microcystis bloom toxic Microcystis multiplex qPCR mcyA mcyB mcyD meTE PCA 

分 类 号：S942.91[农业科学—水产养殖] Q94-4[农业科学—水产科学]