上海交通大学：《微生物学 Microbiology》课程教学资源（双语课件）Lecture 14.2 Methods in Microbial Ecology and Test（MEASURING MICROBIAL ACTIVITIES IN NATURE）

上泽充通大￥ Shanghai Jiao Tong University 1896 1920 1987 2006 Lecture 14-2 Methods in Microbial Ecology CHAPTER22.inag后 BROCK BIOLOGY OF MICROORGANISMS IAO TONG Zhao Liping,Chen Feng School of Life Science and Biotechnology, Shanghai Jiao Tong University Email:cf2001@sjtu.edu.cn

1896 1920 1987 2006 Lecture 14-2 Methods in Microbial Ecology Zhao Liping, Chen Feng School of Life Science and Biotechnology, Shanghai Jiao Tong University Email: cf2001@sjtu.edu.cn CHAPTER 22 in BROCK BIOLOGY OF MICROORGANISMS

泽充通大￥ Shanghai Jiao Tong University 1896 1920 1987 2006 I. MEASURING MICROBIAL ACTIVITIES IN NATURE 风月y 1日9G nnvv

1896 1920 1987 2006 III. MEASURING MICROBIAL ACTIVITIES IN NATURE

上游充廷大￥ Shanghai Jiao Tong University Methods for activity measurement Radioisotopes放射性同位素 Microelectrodes微电极 Stable isotope analyses稳定同位素分析 Microbiology.Shanghai Jiao Tong University

Microbiology, Shanghai Jiao Tong University Methods for activity measurement Radioisotopes放射性同位素 Microelectrodes微电极 Stable isotope analyses 稳定同位素分析

浒充通大学 上 Shanghai Jiao Tong University 14.8 Chemical Assays,Radioisotopes, Microelectrodes In many studies,direct chemical measurements are sufficient Higher sensitivity can be achieved with radioisotopes Proper killed cell controls must be used Microbiology.Shanghai Jiao Tong University

Microbiology, Shanghai Jiao Tong University 14.8 Chemical Assays, Radioisotopes, & Microelectrodes In many studies, direct chemical measurements are sufficient • Higher sensitivity can be achieved with radioisotopes • Proper killed cell controls must be used

Sulfate reduction Photosynthesis Formalin-killed control 坚 Light Microbial activity H2S measurements Killed Lactate Dark Time Time (a)Chemical assay (b)Photosynthesis measured in natural seawater with 14C02 Sulfate reduction 14C-Glucose respiration H2 present 圣 uolnJona Killed、 H2 absent Killed Time Time (c)Sulfate reduction in mud (d)Production of 14CO2 from 14C- 0measured with 35S02- glucose

Chemical assay Photosynthesis measured in natural seawater with 14CO2 Sulfate reduction in mud measured with 35SO4 2- Production of 14CO2 from 14C￾glucose Microbial activity measurements

上帝充通大学 Shanghai Jiao Tong University 14.8.1 Radioisotopes放射性同位素 ©Advantages When 1)very high sensitivity is required,or 2) turnover rates need to be determined,or 3)the fate of portions of a molecule need to be followed, radioisotopes are very useful. @Importance of a proper controli设置对照很重要 To exclude the possibility of chemical transformation排除化 学转化的可能性 ·Killed cell control采用死细胞为对照 ·4%formalin is commonly used.福尔马林 Microbiology.Shanghai Jiao Tong University

Microbiology, Shanghai Jiao Tong University 14.8.1 Radioisotopes 放射性同位素 Advantages • When 1) very high sensitivity is required, or 2) turnover rates need to be determined, or 3) the fate of portions of a molecule need to be followed, radioisotopes are very useful. Importance of a proper control设置对照很重要 • To exclude the possibility of chemical transformation排除化 学转化的可能性 • Killed cell control采用死细胞为对照 • 4% formalin is commonly used.福尔马林

上泽克通大学 Shanghai Jiao Tong University 14.8.2 Microelectrodes微电极 Microelectrodes for activities in microenvironments 用于微环境的活性测定 Measurements taken every 50-100um Chemical species that can be measured by electrodes pH 02 N,O(nitrous oxide) C02 、H2 ·H2S Microbiology.Shanghai Jiao Tong University

Microbiology, Shanghai Jiao Tong University 14.8.2 Microelectrodes 微电极 Microelectrodes for activities in microenvironments可 用于微环境的活性测定 • Measurements taken every 50–100mm Chemical species that can be measured by electrodes • pH • O2 • N2O (nitrous oxide) • CO2 • H2 • H2S

Figure 22.19 Gold Glass Platinum 5μm (a) Membranes Glass 50-100μm NO3 NO 2e1N20+H20→N2+2OH Bacteria Cathode Nutrient solution (b) 2012 Pearson Education,Inc

Figure 22.19 Gold Glass Platinum 5 mm Membranes Glass Bacteria Cathode Nutrient solution 50–100 mm NO3  N2O 2e 1 N2O  H2O N2  2 OH © 2012 Pearson Education, Inc

2 pH 0 12 H2S paeM'W PIne 3 52ot1irekaegy时leorgns1ie 6 7 8 9 pH Microprofiles of O2, H,S and pH in a 200 400 600 800 1000 hot spring 02 or H2S (M) microbial mat Figure 18-19b Brock Biology of Microorganisms 11/e 2006 Pearson Prentice Hall,Inc. Activity research in microbial mats(layered microbial communities)微生物垫（多层次微生物生态系统)

Microprofiles of O2 , H2S and pH in a hot spring microbial mat Activity research in microbial mats (layered microbial communities)微生物垫（多层次微生物生态系统）

02 pH 0 12 H2S paeM'W pIne 3 6 7 8 9 pH 0 200 400 600 8001000 02 or H2S (M) Microbial mats Figure 18-19b Brock Biology of Microorganisms 11/e 2006 Pearson Prentice Hall,Inc. -Cyanobacteria:uppermost layers -Anoxygenic phototrophic bacteria:subsequent layers -Chemoorganotrophic bacteria:lower layers -Balance between photosynthesis and decomposition

Cyanobacteria:uppermost layers Anoxygenic phototrophic bacteria: subsequent layers Chemoorganotrophic bacteria: lower layers Balance between photosynthesis and decomposition Microbial mats