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鱼类环境DNA保存条件的比较研究
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作者 刘洋 应方 +3 位作者 杨俊 王焕英 洪文杰 王庭璋 《生物学杂志》 CAS CSCD 北大核心 2024年第3期114-120,共7页
为探索环境水样中鱼类DNA的最佳保存方法和保存时间,选取饲养鱼的水样为研究对象,在无核酸污染的环境中,采用0.45μm孔径滤膜对相同体积的水样进行预处理,并对富集环境DNA(eDNA)的滤膜进行不同保存方法和不同保存时间捕获的eDNA质量、鱼... 为探索环境水样中鱼类DNA的最佳保存方法和保存时间,选取饲养鱼的水样为研究对象,在无核酸污染的环境中,采用0.45μm孔径滤膜对相同体积的水样进行预处理,并对富集环境DNA(eDNA)的滤膜进行不同保存方法和不同保存时间捕获的eDNA质量、鱼类12S rRNA基因文库浓度、基于ASVs方法的鱼类12S rRNA扩增子测序结果的比较研究。研究结果显示:-20℃冷冻、-80℃冷冻、无水乙醇-常温、75%乙醇-常温、Longmire’s保存液-常温以及TK保存液-常温等6种保存方法捕获的eDNA效果最好的是TK保存液-常温,其次为冷冻保存(-20℃、-80℃)。-20℃冷冻、-80℃冷冻和TK保存液-常温等3种保存方法在同一保存时间下具有良好的稳定性,且在保存时间2周内的测试条件下,也能稳定保存滤膜,最大程度还原水样中鱼类的遗传信息。研究对eDNA样本最佳保存条件的探索,将为第三方基因检测实验室对鱼类eDNA保存策略提供技术基础,便于eDNA宏条形码技术的开展。 展开更多
关键词 EDNA 保存条件 DNA提取 扩增子测序 asvs方法
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Diversity and geographical distribution of haptophyte Phaeocystis in the Chinese seas based on metabarcoding analysis
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作者 Qingshang SONG Yue XUE +2 位作者 Yanying ZHANG Jiehui YIN Pingping SHEN 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2023年第6期2197-2207,共11页
Phaeocystis is an important bloom-forming species and over 100 blooms have occurred since 1997 along the Chinese coasts,while only the species P.globosa was described.In this project,a total of 246 phytoplankton sampl... Phaeocystis is an important bloom-forming species and over 100 blooms have occurred since 1997 along the Chinese coasts,while only the species P.globosa was described.In this project,a total of 246 phytoplankton samples collected from the Bohai Sea,the Yellow Sea,and the East China Sea in April 2021 were analyzed to evaluate the species diversity and geographical distribution of the genus Phaeocystis based on metabarcoding 18S r DNA sequence.Four described Phaeocystis species were recognized,including P.globosa,P.pouchetii,P.jahnii,and P.cordata,and each has distinguished geographical distribution characteristics.P.globosa was the most widespread and abundant species,and P.pouchetii was found in the Bohai Sea and the northern Yellow Sea with substantially elevated abundance.P.cordata was also a widespread species,but its abundance was relatively low,while P.jahnii gathered only in the southeastern East China Sea.Water temperature,phosphate as well as ammonium contents were found to be associated with the abundance of P.globosa,P.pouchetii,and P.jahnii significantly.Moreover,two uncharacterized Phaeocystis species were detected in the Chinese seas,indicating the diversity of the genus remains underestimated worldwide. 展开更多
关键词 HAPTOPHYTA PHAEOCYSTIS species diversity distribution Chinese seas amplicon sequence variant
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有害藻华的宏条形码分析:机会与挑战 被引量:11
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作者 陈楠生 《海洋科学》 CAS 北大核心 2020年第7期116-134,共19页
宏条形码分析(metabarcoding analysis)是通过对野外样本的分子标记(即条形码)进行PCR扩增、测序,并利用测序结果分析野外样本物种组成、物种相对丰度,及其时空动态变化。这项在近20年里逐步发展起来的分子技术,得益于DNA测序技术的快... 宏条形码分析(metabarcoding analysis)是通过对野外样本的分子标记(即条形码)进行PCR扩增、测序,并利用测序结果分析野外样本物种组成、物种相对丰度,及其时空动态变化。这项在近20年里逐步发展起来的分子技术,得益于DNA测序技术的快速发展,包括测序通量的提升和测序成本的降低,以及生物信息学分析方法的快速开发。基于可操作分类单元(operational taxonomic units,OTUs)分析方法,特别是最近的扩增子序列变异(amplicon sequence variants,ASVs)方法的开发,极大促进了宏条形码分析的应用。在世界及我国的海洋生态领域,宏条形码分析的应用还处于起步阶段。随着分析方法的日趋成熟,可适用于不同应用领域的分子标记不断增加,以及参考数据库的不断完善,宏条形码分析将释放出广阔的应用潜力。可以预期,宏条形码分析将在系统分析我国近岸海域有害藻华物种(及其他浮游生物物种)的组成和相对丰度中起关键作用,通过发现新物种,辨别隐存种,跟踪物种的生物地理学,进而帮助解析有害藻华暴发机理。 展开更多
关键词 有害藻华物种 分子标记 可操作分类单元 扩增子序列变异 宏条形码分析 生物地理学
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Microbiome Diversity Analysis of the Bacterial Community in Idah River, Kogi State, Nigeria
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作者 Deborah Ebunoluwa Adedire Adijat Ozohu Jimoh +10 位作者 Zainab Kashim-Bello Bilyaminu Adam Wase Shuaibu Oluwabukola Atinuke Popoola Kabiru Isa Pate Onyinye Stainless Uzor Eno Etingwa Jumoke Florence Joda Oyetola Olanrewaju Opaleye Victoria Adejoke Ogunlowo Kehinde Rukayyat Adeniran Oyekanmi Nashiru 《Advances in Microbiology》 2022年第5期343-362,共20页
The analysis of bacterial diversity in aquatic systems particularly in rivers, lakes, and streams can provide useful data on the effect of anthropogenic activities on such water bodies to humans and fishes. Idah River... The analysis of bacterial diversity in aquatic systems particularly in rivers, lakes, and streams can provide useful data on the effect of anthropogenic activities on such water bodies to humans and fishes. Idah River, the focal point of this study, is an offshoot of the two major Nigerian rivers characterized by observed human activities and pollution sources. Water samples were collected from four designated sites and assessed for their bacterial assemblages and structure, using PacBio Single-Molecule Real-Time (SMRT) sequencing technology. The full length of the 16S rRNA gene was sequenced, and Amplicon Sequence Variants were generated using the DADA2 workflow optimised for PacBio long-read amplicons in Rstudio. A total of 8751 high-quality reads obtained were taxonomically classified as 24 phyla, 42 classes, 84 orders, 125 families, 156 genera, and 106 species. Taxonomical composition revealed Proteobacteria as the most abundant phyla across all sample sites. At the genera level, Azospira (57.03%) was the most dominant ASV in Docking Point A, while Acinetobacter (66.67%) was the most abundant ASV in Docking Point B. In Idah Axis Confluence, hgcl clade (65.66%) was the most prevalent ASV, whereas Holophaga (42.86%) was the most common ASV in Idah Axis Midstream. Genera analysis also revealed that 12.9% of the total ASVs were discovered across all sample sites. Among these were pathogenic bacteria, reducers, and degraders of domestic and animal wastes. Observed results provide evidence that sampled sites of Idah River are contaminated, most likely through constant human activities and thus, could have an impact on resident fishes as well. This study, therefore, agrees with a previous report from the river, which used standard microbial procedures. However, next-generation sequencing techniques employed revealed more bacterial community than the former, including unresolved taxonomic sequences that may be novel. 展开更多
关键词 Idah River Dada2 PacBio Sequencing Bacterial Diversity amplicon sequence variants
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Main environmental drivers of abundance,diversity and community structure of comammox Nitrospira in paddy soils 被引量:1
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作者 Fuyun GAO Yaying LI +2 位作者 Haoxin FAN Jiantao XUE Huaiying YAO 《Pedosphere》 SCIE CAS CSCD 2023年第5期808-818,共11页
The recently discovered complete ammonia oxidizers comammox Nitrospira contain clades A and B that can establish an independent one-step nitrification process;however,little is known about their environmental drivers ... The recently discovered complete ammonia oxidizers comammox Nitrospira contain clades A and B that can establish an independent one-step nitrification process;however,little is known about their environmental drivers or habitat distributions in agricultural soils.Previous studies on comammox Nitrospira in paddy soils have mainly focused on small-scale samples,and there is a lack of multisite research on comammox Nitrospira in paddy soils.In this study,we conducted a survey of 36 paddy soils to understand the community structure,abundance,and diversity of comammox Nitrospira and the degree to which they are affected by environmental factors at a large scale.Comammox Nitrospira were found to be widely distributed among the paddy soils.The abundance of comammox Nitrospira clade A was mostly lower than that of clade B,whereas its diversity was mostly higher than that of clade B.Correlation analysis showed that multiple factors affected(P<0.05)the abundance of comammox Nitrospira,including soil pH,organic matter,total carbon,and total nitrogen,latitude,mean annual temperature,and mean annual precipitation.Moreover,there was a clear relationship between the comammox Nitrospira community and habitat,indicating that some amplicon sequence variants(ASVs)had a unique dominant position in specific habitats.Phylogenetic analysis showed that the ASVs of comammox Nitrospira clade A clustered with the known sequences in the paddy soils and were significantly different from the known sequences in other habitats,which may be related to the unique paddy field habitat.In contrast,comammox Nitrospira clade B showed no clear habitat dependence.These results support the wide distribution and high abundance of comammox Nitrospira in paddy soils and provide novel insights into nitrogen cycling and nutrient management in agricultural ecosystems. 展开更多
关键词 ammonia-oxidizing archaea ammonia-oxidizing bacteria amplicon sequence variant complete ammonia oxidizer environmental factor HABITAT NITRIFICATION nitrogen cycling
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