基于聚球藻报告基因的水体营养盐生物可利用性检测  

DETECTION OF NUTRIENT BIOAVAILABILITY IN WATER BODIES BASED ON SYNECHOCOCCUS REPORTER GENES

作　　者：刘浩[1] 韦嘉宁 王鑫威[1] 姜海波 LIU Hao;WEI Jia-Ning;WANG Xin-Wei;JIANG Hai-Bo(School of Marine Sciences,Ningbo University,Ningbo 315211,China;Southern Marine Science and Engineering Laboratory(Zhuhai),Zhuhai 519080,China)

机构地区：[1]宁波大学海洋学院,宁波315211 [2]南方海洋科学与工程广东省实验室(珠海),珠海519080

出　　处：《水生生物学报》2023年第12期1899-1908,共10页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金(42188102、32170108和91951111);南方海洋科学与工程广东省实验室(珠海)自主科研项目(SML2021SP204);宁波市重点研发计划(2022Z189)资助。

摘　　要：研究从生物可利用性角度出发,筛选出2个高效且特异响应氮、磷生物可利用度的聚球藻启动子,驱动细菌荧光素酶报告基因,构建了用于直接检测环境样品中营养盐生物可利用度的微藻细胞生物报告器,获得了生物发光与标准氮(NO_(3)^(-))、磷(PO_(4)^(3-))营养盐浓度的关系曲线,可用于水体氮、磷生物可利用度的快速检测。同时,使用国标法与本方法对南海航次水样进行了测定,国标法测得的磷浓度为0.17—3.00μmol/L,氮浓度为1.04—49.34μmol/L,而本方法测定的磷浓度为0.10—1.13μmol/L,与国标法测定的结果接近;氮浓度为32.57—98.06μmol/L,比国标法测得的浓度高,可能是由于水样中还含有其他形式的氮。研究提出了利用生物报告器检测并量化海水氮、磷生物可利用性的方法,是对水体营养盐化学浓度指标的重要补充,为环境样品营养盐生物可利用性检测提供了新的方向和思路。Nitrogen and phosphorus nutrients are the basic components of marine phytoplankton and key factors in controlling marine primary productivity.Previous studies have focused on the determination of nutrient chemical concentrations and neglected the detection of the actual bioavailability of nutrients in the water column.In order to achieve the detection of nitrogen and phosphorus nutrient bioavailable concentrations,this study screened two efficient and specific response to nitrogen and phosphorus bioavailable promoters of Synechococcus,drove the expression of bacterial luciferase reporter genes,constructed a microalgal cell bioreporter for the direct detection of nutrient bioavailability in environmental samples,and obtained the relationship curves between bioluminescence and standard nitrogen (NO_(3)^(-)) and phosphorus (PO_(4)^(3-)) nutrient concentrations,which can be used for the rapid detection of nitrogen and phosphorus bioavailability in water bodies.Meanwhile,the water samples of the South China Sea cruise were measured using the national standard method and the present method,and the phosphorus concentration measured by the national standard method was 0.17—3.00μmol/L and the nitrogen concentration was 1.04—49.34μmol/L,while the phosphorus concentration measured by the present method was 0.10—1.13μmol/L,which was close to the results determined by the national standard method;the nitrogen concentration was 32.57—98.06μmol/L,which was different from the results determined by the national standard,probably because the water samples also contained other forms of nitrogen.The study proposes a method to detect and quantify the bioavailability of seawater nitrogen and phosphorus using biological reporters,which is an important complement to the chemical concentration index of nutrients in water bodies and provides new directions and ideas for the detection of nutrient bioavailability of environmental samples.

关 键 词：营养盐检测 生物可利用性 生物报告器 浮游植物 

分 类 号：Q178.1[生物学—水生生物学]