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Related Concept Videos

Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

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Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
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Applications of Molecular Taxonomy01:20

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Molecular taxonomy has revolutionized the understanding and classification of bacteria, providing precise insights into their diversity, evolutionary relationships, and ecological roles. By utilizing molecular techniques such as DNA sequencing and fingerprinting, researchers have made significant strides in various fields related to bacterial studies.Resolving Taxonomic AmbiguitiesMolecular taxonomy has been instrumental in distinguishing closely related bacterial species initially thought to...
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Related Experiment Video

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Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing
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Pseudomonas-specific 16S rRNA insect gut-microbiome profiling using next-generation sequencing.

Ayushi Gupta1, Suresh Nair1

  • 1Plant-Insect Interaction Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110067, India.

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|December 17, 2022
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Summary

This study details a Pseudomonas-specific gut microbiome profiling protocol for brown planthopper (BPH) populations using next-generation sequencing and qPCR. The method enables comparative analysis of Pseudomonas species composition and titers in BPH.

Keywords:
GenomicsImmunologyMicrobiologyMolecular BiologySequencing

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Area of Science:

  • Microbiology
  • Entomology
  • Genomics

Background:

  • The gut microbiome of insects, like the brown planthopper (BPH), plays a critical role in their adaptation and survival.
  • Understanding the microbial communities, particularly specific bacterial groups like Pseudomonas, is essential for pest management strategies.
  • Environmental factors such as climate change and geographical distribution can influence insect-associated microbial communities.

Purpose of the Study:

  • To present a detailed protocol for Pseudomonas-specific 16S rRNA gut-microbiome profiling in brown planthopper (BPH) populations.
  • To establish a method for comparative analysis of Pseudomonas species structure and composition across diverse BPH populations.
  • To quantify Pseudomonas species titers in BPH using quantitative polymerase chain reaction (qPCR).

Main Methods:

  • Next-generation sequencing (NGS) for 16S rRNA gene profiling of the gut microbiome.
  • Development of a Pseudomonas-specific profiling technique.
  • Quantitative polymerase chain reaction (qPCR) for accurate enumeration of Pseudomonas titers.

Main Results:

  • A comprehensive protocol for analyzing the Pseudomonas gut microbiome in BPH populations was successfully developed.
  • The protocol allows for comparative analysis of Pseudomonas species composition and structure across different geographical locations and climatic conditions.
  • qPCR enabled the quantification of Pseudomonas species abundance within BPH individuals.

Conclusions:

  • The presented protocol provides a robust method for investigating Pseudomonas gut-microbiome dynamics in BPH.
  • This approach can be adapted for microbiome analysis and population monitoring of other pest species.
  • Understanding these microbial dynamics is crucial for biodiversity assessment, speciation studies, and pest adaptation research.