Direct sequencing of insect symbionts via nanopore adaptive sampling

Badger, Jonathan H, Giordano, Rosanna, Zimin, Aleksey, Wappel, Robert, Eskipehlivan, Senem M, Muller, Stephanie, Donthu, Ravikiran, Soto-Adames, Felipe, Vieira, Paulo, Zasada, Inga, Goodwin, Sara (February 2024) Direct sequencing of insect symbionts via nanopore adaptive sampling. Current Opinion in Insect Science, 61. p. 101135. ISSN 2214-5745

URL: https://www.ncbi.nlm.nih.gov/pubmed/37926187
DOI: 10.1016/j.cois.2023.101135

Abstract

Insect symbionts can alter their host phenotype and their effects can range from beneficial to pathogenic. Moreover, many insects exhibit co-infections, making their study more challenging. Less than 1% of insect species have high-quality referenced genomes available and fewer still also have their symbionts sequenced. Two methods are commonly used to sequence symbionts: whole-genome sequencing to concomitantly capture the host and bacterial genomes, or isolation of the symbiont's genome before sequencing. These methods are limited when dealing with rare or poorly characterized symbionts. Long-read technology is an important tool to generate high-quality genomes as they can overcome high levels of heterozygosity, repeat content, and transposable elements that confound short-read methods. Oxford Nanopore (ONT) adaptive sampling allows a sequencing instrument to select or reject sequences in real time. We describe a method based on ONT adaptive sampling (subtractive) approach that readily permitted the sequencing of the complete genomes of mitochondria, Buchnera and its plasmids (pLeu, pTrp), and Wolbachia genomes in two aphid species, Aphis glycines and Pentalonia nigronervosa. Adaptive sampling is able to retrieve organelles such as mitochondria and symbionts that have high representation in their hosts such as Buchnera and Wolbachia, but is less successful at retrieving symbionts in low concentrations.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > analysis and processing
bioinformatics
bioinformatics > genomics and proteomics
organism description > animal
organism description > animal > insect
CSHL Authors:
Communities: CSHL labs > McCombie lab
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 1 February 2024
Date Deposited: 16 Jan 2024 15:11
Last Modified: 16 Feb 2024 16:48
Related URLs:
URI: https://repository.cshl.edu/id/eprint/41401

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