Evidence map›Paper›PMID 41728922›Full record

ArticleFEMS microbiology ecology2026

Optimizing primer selection for microbial diversity studies in freshwater lakes: a comprehensive evaluation of 16S rRNA, 18S rRNA, and ITS gene regions.

Chen Wang, Xinyu Chen, Yuxuan Lei, Zhen Shen, Yi Gong, Hai Xu, Xiangming Tang

Abstract read
In one paragraph

Article in FEMS microbiology ecology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Article
  2. Review
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Chen WangTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.ORCID 0009-0007-0969-6947
Xinyu ChenTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.
Yuxuan LeiTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.
Zhen ShenTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.
Yi GongTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.
Hai XuTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.
Xiangming TangTaihu Laboratory for Lake Ecosystem Research, State Key Laboratory of Lake and Watershed Science for Water Security, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 211135, China.ORCID 0000-0002-2816-1004

Funding

Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences NIGLAS2022GS03National Natural Science Foundation of China 42371065
6 · The paper itself

Abstract

Accurate assessment of microbial community structure via high-throughput sequencing is fundamentally dependent on primer selection; however, systematic primer optimization for freshwater lake ecosystems remains insufficiently explored. Using the representative Lake Qiandao as a model, this study provides an integrative comparison of multiple primer sets targeting the 16S rRNA gene (V1-V2, V3-V4, V4, and V5-V7), the 18S rRNA gene (V4-1 and V4-6), and the Internal Transcribed Spacer (ITS1 and ITS2) regions. Our results elucidate that the V3-V4 region yielded significantly higher bacterial diversity and a more biologically representative community structure. For eukaryotic microorganisms, the V4-1 primers exhibited superior performance, capturing a broader taxonomic spectrum of algae (e.g. Ochrophyta and Cryptophyta) and protozoan groups. Regarding fungi, the ITS2 region provided enhanced stability and more comprehensive taxonomic coverage compared to ITS1. This study establishes a standardized molecular framework for microbial diversity research in freshwater limnology and underscores the necessity of primer optimization to ensure the high-fidelity resolution of microbial community structures and their associated ecological function.

Indexed as

BacteriaDNA PrimersLakesRNA, Ribosomal, 16SRNA, Ribosomal, 18SBiodiversityDNA, Ribosomal SpacerFungiMicrobiotaPhylogenySequence Analysis, DNADNA PrimersDNA, Ribosomal SpacerRNA, Ribosomal, 16SRNA, Ribosomal, 18S16S rRNA18S rRNAfreshwater lakesITSmicrobial diversityprimer selection

Identifiers

PMID41728922
PMCPMC12934351

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.