引用本文:	王建龙,侯天琪,王欣欣,庄昀筠,陈洪举,刘光兴.固定剂和保存条件对桡足类及其肠道内容物DNA质量的影响[J].海洋科学,2025,49(12):104-114.

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固定剂和保存条件对桡足类及其肠道内容物DNA质量的影响
王建龙¹, 侯天琪¹, 王欣欣¹, 庄昀筠^1,2, 陈洪举^1,2, 刘光兴^1,2
1.中国海洋大学环境科学与工程学院/海洋环境与生态教育部重点实验室, 山东青岛 266100;2.青岛海洋科技中心海洋生态与环境科学功能实验室, 山东青岛 266237

摘要:

DNA条形码技术已广泛应用于桡足类多样性及其食物组成研究,是解析浮游生物群落结构和海洋食物网动态的有力工具。样本的有效保存是开展PCR扩增和基因测序等DNA下游分析的关键前提。本研究评估了不同固定剂与保存时长对桡足类及其肠道内容物DNA含量和质量的影响,并通过qPCR定量不同长度的18S rDNA片段以评估DNA的完整性。结果显示, RNAlater是保存桡足类DNA的最佳固定剂,在2个月保存期内能够维持最高的DNA含量和完整性;乙醇对肠道内容物DNA的固定和保存效果最佳,表明其具有良好的穿透性,可迅速抑制桡足类的生理活动及肠道微生物的代谢活性。中性鲁哥氏液亦可即时或短期固定桡足类肠道内容物DNA,但不宜于长期保存。本研究结果为桡足类及其现场食物组成研究采样策略制定、固定方法选择及保存条件优化提供了科学依据,建议根据研究对象优先级(桡足类或肠道内容物DNA)选择相应的固定剂和保存条件。

关键词: 桡足类肠道内容物固定剂保存条件 DNA

DOI：10.11759/hykx20250430002

分类号:P735

基金项目:崂山实验室科技创新项目(LSKJ202202804);国家自然科学基金项目(41876156)

Effects of the fixative and storage duration on the quality of DNA obtained from copepods and their gut contents

WANG Jianlong¹, HOU Tianqi¹, WANG Xinxin¹, ZHUANG Yunyun^1,2, CHEN Hongju^1,2, LIU Guangxing^1,2

1.Key Laboratory of Marine Environment and Ecology, Ministry of Education, College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China;2.Laboratory for Marine Ecology and Environmental Science, Qingdao Marine Science and Technology Center, Qingdao 266237, China

Abstract:

The DNA barcoding technique has been widely used in studying the diversity of copepods and their diet in situ, serving as a powerful tool for understanding the plankton community structure and marine food web dynamics. Effective sample preservation is a critical prerequisite for downstream DNA analyses, including PCR amplification and gene sequencing. In this study, we evaluated the effects of the fixative and storage duration on both the yield and quality of DNA extracted from copepods and their gut contents. DNA integrity was assessed through the qPCR quantification of 18S rDNA fragments with different lengths. Our results showed that RNAlater exhibited optimal performance in fixing and preserving copepod DNA with the highest yield and integrity over a two-month storage period. Ethanol demonstrated the best efficacy in the fixation and preservation of gut content DNA as it exhibits excellent permeability and rapidly terminates the physiological and metabolic activities of copepods and gut microbiota, respectively. Neutral Lugol's solution can achieve immediate or short-term fixation of gut content DNA; however, it is unsuitable for long-term preservation. These findings provide insights into sampling strategies, fixation protocols, and storage conditions for investigating copepods and their diet in situ. They guide the use of appropriate fixatives and preservation conditions based on research priorities(copepod DNA or gut content DNA).

Key words: copepod gut content fixation storage DNA