荧光定量PCR(qPCR)检测“漂浮型”浒苔微观繁殖体  

Real-Time Fluorescence Quantitative PCR(qPCR)Detection of Micro-Propagules of“Floating Ecotype”Ulva prolifera

作　　者：张宝堂 肖洁 马晓君 缪晓翔 李梅 范士亮 臧宇 张学雷 王宗灵 ZHANG Baotang;XIAO Jie;MA Xiaojun;MIAO Xiaoxiang;LI Mei;FAN Shiliang;ZANG Yu;ZHANG Xuelei;WANG Zongling(First Institute of Oceanography,MNR,Qingdao 266061,China;Key Lab of Science and Engineering for Marine Ecological Environment,MNR,Qingdao 266061,China;Laboratory of Marine Ecology and Environmental Sciences,Laoshan Laboratory,Qingdao 266037,China)

机构地区：[1]自然资源部第一海洋研究所,山东青岛266061 [2]自然资源部海洋生态环境科学与技术重点实验室,山东青岛266061 [3]崂山实验室海洋生态与环境科学功能实验室,山东青岛266037

出　　处：《海洋科学进展》2025年第1期188-199,共12页Advances in Marine Science

基　　金：崂山实验室科技创新项目(LSKJ202203700);国家重点研发计划项目(2022YFC3106000);国家自然科学基金项目(42276221)。

摘　　要：浒苔绿潮是我国南黄海海域最严重的生态灾害。浒苔微观繁殖体的时空分布、种群动态,是黄海浒苔绿潮业务化监测工作的重要内容,但传统的培养法实验周期长、检测效率低,无法满足快速监测预警的要求。本文利用297 bp的漂浮浒苔特异性DNA片段,研发了浒苔微观繁殖体的荧光定量PCR(qPCR)检测技术,并以人工释放的浒苔配子为研究对象,研究了浒苔配子细胞中目标基因片段的拷贝数(240 copies/cell)。利用qPCR法对2023年4月苏北浅滩海域水样进行检测,并与传统培养法进行比较。结果显示,黄海大规模浒苔绿潮暴发前期,苏北浅滩水体中浒苔微观繁殖体丰度为2.1×10^(7) copies/L(约87602 cells/L,qPCR)和6.5株/L(传统培养法),且呈现近岸(靠近筏架区)高、离岸低的趋势。两种方法对浒苔微观繁殖体的检出效率和揭示的分布规律基本一致。qPCR法灵敏、高效,可用于黄海浒苔绿潮业务化监测工作。The Ulva prolifera green tide is one of the most severe ecological disasters in the Southern Yellow Sea.The spatio-temporal distribution and population dynamics of Ulva prolifera micro-propagules are important parts of the regular monitoring of green tide in the Yellow Sea.However,the traditional culture method has long experimental period and low detection efficiency,which cannot satisfy the requirement of rapid monitoring and early warning.In this paper,a fluorescence quantitative PCR(qPCR)technique based on one 297 bp DNA fragment specific to the floating Ulva prolifera was developed to detect the Ulva prolifera micro-propagules.The artificially released Ulva prolifera gametes were used to study the copy number of the target gene fragment in Ulva prolifera cells(240 copies/cell).The water samples of Subei Shoal were collected in April 2023 and the abundances of Ulva prolifera micro-propagules were screened based on both qPCR and traditional culturing methods.The results showed that the average abundances of Ulva prolifera micro-propagules were 2.1×10^(7) copies/L(approximately 87602 cells/L)and 6.5 inds/L by qPCR and traditional culturing method,respectively,and the abundances were higher in the near-shore water(around raft region)and lower in off-shore.The detection efficiency and the revealed distribution patterns of the Ulva prolifera micro-propagules were consistent through the two methods.This qPCR method is sensitive and efficient,and can be used for the regular monitoring on Ulva prolifera green tide in the Yellow Sea.

关 键 词：微观繁殖体 荧光定量PCR 绿潮 浒苔 黄海 

分 类 号：P714.5[天文地球—海洋科学]