地球科学进展 ›› 2021, Vol. 36 ›› Issue (12): 1247 -1257. doi: 10.11867/j.issn.1001-8166.2021.118

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活体底栖有孔虫鉴别方法及其应用
张涵 1 , 2 , 3( ), 许博超 1 , 2( ), 郭肖伊 1 , 2 , 3, 张晓洁 1 , 2 , 3, 于志刚 1 , 2   
  1. 1.深海圈层与地球系统前沿科学中心,海洋化学理论与工程技术教育部重点实验室,中国海洋大学,山东 青岛 266100
    2.青岛海洋科学与技术试点国家实验室,海洋生态与环境科学功能实验室,山东 青岛 266071
    3.中国海洋大学 化学化工学院,山东 青岛 266100
  • 收稿日期:2021-08-09 修回日期:2021-11-22 出版日期:2021-12-10
  • 通讯作者: 许博超 E-mail:zhhan926@163.com;xubc@ouc.edu.cn
  • 基金资助:
    国家自然科学基金项目“多核素示踪海底地下水排放对长江口外低氧形成的贡献”(41876075);高校基本科研业务费中国海洋大学研究生自主科研项目“有孔虫壳体Mn/Ca比值对长江口季节性低氧现象的指示研究”(201961012)

Methods and Applications for Identifying Living Benthic Foraminifera

Han ZHANG 1 , 2 , 3( ), Bochao XU 1 , 2( ), Xiaoyi GUO 1 , 2 , 3, Xiaojie ZHANG 1 , 2 , 3, Zhigang YU 1 , 2   

  1. 1.Frontiers Science Center for Deep Ocean Multispheres and Earth System,Key Laboratory of Marine Chemistry Theory and Technology,Ministry of Education,Ocean University of China,Qingdao 266100,China
    2.Laboratory for Marine Ecology and Environmental Science,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266071,China
    3.College of Chemistry and Chemical Engineering,Ocean University of China,Qingdao 266100,China
  • Received:2021-08-09 Revised:2021-11-22 Online:2021-12-10 Published:2022-01-20
  • Contact: Bochao XU E-mail:zhhan926@163.com;xubc@ouc.edu.cn
  • About author:ZHANG Han (1998-), female, Taian County, Shandong Province, Ph.D student. Research area include micropalaeontology. E-mail: zhhan926@163.com
  • Supported by:
    the National Natural Science Foundation of China "Using multi-isotopes to study the contribution of Submarine Groundwater Discharge to hypoxia formation off the Yangtze River Estuary"(41876075);The Fundamental Research Funds for the Central Universities, Independent Research Project for Graduate Students of Ocean University of China "Using benthic foraminiferal Mn/Ca ratio as a proxy for coastal seasonal hypoxia in the Changjiang Estuary"(201961012)

活体底栖有孔虫在底栖生态系统的环境指示研究等诸多研究领域都极具应用价值,但如何准确快速有效地识别活体有孔虫具有挑战性。目前,研究人员已经建立了多种方法用于识别活体底栖有孔虫。在调研相关研究的基础上,对这些方法的优缺点及其应用中的注意事项进行了总结。虎红染色法操作快速简便、实验成本低,是最为常用的活体有孔虫鉴别方法。苏丹黑B染色法和MTT染色法比虎红染色法准确性更高,但操作略显复杂,在野外调查研究中应用较少。荧光探针法不影响有孔虫生命活动且准确性更高,但其操作复杂、成本较高,常被推荐用于低氧/缺氧条件下的有孔虫研究。三磷酸腺苷分析法和超显微结构法均具有较高的准确性,但操作复杂、设备要求高,前者被广泛应用于活体有孔虫数量密度的估算和生物量的计算,并被推荐用于低氧/缺氧条件下的有孔虫相关研究,后者主要用于分析评价有孔虫对缺氧、重金属污染等环境压力的适应性。研究者应根据研究目的、研究成本等条件选择一种或多种合适的研究方法,以获得更为可靠的研究结果。

Living benthic foraminifera have significant values in many research fields, such as indication of environmental factors of benthic ecosystem. In the study of benthic foraminifera, one of the most controversial issues is how to identify living ones accurately and quickly. Researchers have developed a variety of methods for identifying living foraminifera. Here, we summarized the advantages and disadvantages and cautions during their applications. Rose Bengal is relatively quick and inexpensive, becoming the most commonly used method for identifying living foraminifera. Sudan black B and MTT are more accurate than Rose Bengal, but their operation is slightly complicated and often overlooked as a method to distinguish foraminifera. Fluorogenic probes are not harmful to foraminiferal living activities and have higher accuracy, but complicated and expensive. They are often recommended for studies of foraminifera under hypoxic/anoxic conditions. Adenosine Triphosphate Assay and Ultrastructural Studies have high accuracy, but the operation is complicated and the equipment requirements are high. Adenosine Triphosphate Assay is widely used in estimating numerical density of living foraminifera and calculation of foraminifera biomass, and also recommended for the study of foraminifera under hypoxic/anoxic conditions. Ultrastructural Studies can analyze the adaptability of foraminifera to specific environmental conditions such as hypoxia and heavy metal pollution. Researchers should choose one or more appropriate methods according to research objectives, research costs and other conditions to obtain more reliable research results.

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表1 活体底栖有孔虫鉴别的方法原理、优缺点及其应用
Table 1 Principles, advantages and disadvantages, application methods for identifying living benthic foraminifera
方法 原理 优点 缺点 应用
虎红染色法 虎红染色剂与蛋白质分子发生超分子显色反应形成玫瑰红色的超分子复合物 设备简单;操作快速;成本低 可能产生“假阳性”的结果;不透明有孔虫的染色结果难以确认 最为常用的活体有孔虫鉴别方法
苏丹黑B 染色法 苏丹黑B染色剂溶解于细胞内的含脂结构使脂类物质呈现棕黑或深黑色 比虎红染色结果更为可靠 可能产生“假阳性”的结果;不透明有孔虫的染色结果难以确认 应用较少
MTT染色法 MTT被活细胞内的酶促反应转化为红紫色结晶甲臜并沉积在细胞中 比虎红染色结果更为可靠 需要在检查样品采集海水温度下活体和死亡有孔虫之间显色的差异 应用较少
荧光探针法 CTG被活细胞内的非特异性酯酶分解产生荧光物质,以特定的波长激发显示绿色荧光 准确性高;不会对活体有孔虫的生命活动产生不良影响 试剂设备昂贵;操作略为繁琐 低氧/缺氧条件下的有孔虫 研究
ATP分析法 生物死亡后细胞内的三磷酸腺苷能够迅速发生去磷酸化反应 与染色法相比具有更高的准确性 操作复杂、设备要求高;难以确定活体与死体之间的精确界限 估算活体有孔虫的数值密度,评估有孔虫生物量;低氧/缺氧条件下的有孔虫研究
超显微结构法 有孔虫细胞内细胞器的外观、丰度和组织结构等超显微结构可作为细胞生理状态的指标 具有较高的可靠性 操作复杂且耗时;专业知识和操作技能要求高;不适用于壳体较厚的多房室有孔虫 活体有孔虫生理状态研究,分析有孔虫对特定环境条件的适应性
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