Protocol for isolating the single bacteriocyte from whiteflies for single-cell RNA-seq analysis.

STAR Protoc

Liaoning Key Laboratory of Economic and Applied Entomology, College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China; Shenyang Key Laboratory of Surveillance and Management for Vegetable Diseases and Insect Pests, College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China. Electronic address:

Published: March 2025

Bacteriocytes are specialized insect cells adapted to harbor symbionts. However, their low number in individual whiteflies makes obtaining enough for transcriptome sequencing challenging. Here, we present a protocol for the isolation of whitefly bacteriocytes. We describe steps for preparing the single bacteriocyte sample, cDNA amplification library construction, sequencing, and data analysis. Our protocol offers convenience and efficiency for conducting single-cell sequencing for bacteriocytes and has the potential to be applied to other insect cells. For complete details on the use and execution of this protocol, please refer to Li et al..

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.xpro.2025.103691DOI Listing

Publication Analysis

Top Keywords

single bacteriocyte
8
insect cells
8
protocol
4
protocol isolating
4
isolating single
4
bacteriocyte whiteflies
4
whiteflies single-cell
4
single-cell rna-seq
4
rna-seq analysis
4
analysis bacteriocytes
4

Similar Publications

Protocol for isolating the single bacteriocyte from whiteflies for single-cell RNA-seq analysis.

STAR Protoc

March 2025

Liaoning Key Laboratory of Economic and Applied Entomology, College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China; Shenyang Key Laboratory of Surveillance and Management for Vegetable Diseases and Insect Pests, College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China. Electronic address:

Bacteriocytes are specialized insect cells adapted to harbor symbionts. However, their low number in individual whiteflies makes obtaining enough for transcriptome sequencing challenging. Here, we present a protocol for the isolation of whitefly bacteriocytes.

View Article and Find Full Text PDF

A complete DNA repair system assembled by two endosymbionts restores heat tolerance of the insect host.

Proc Natl Acad Sci U S A

December 2024

State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.

DNA repair systems are essential to maintain genome integrity and stability. Some obligate endosymbionts that experience long-term symbiosis with the insect hosts, however, have lost their key components for DNA repair. It is largely unexplored how the bacterial endosymbionts cope with the increased demand for mismatch repairs under heat stresses.

View Article and Find Full Text PDF

Deciphering deep-sea chemosynthetic symbiosis by single-nucleus RNA-sequencing.

Elife

August 2024

Center of Deep-Sea Research, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.

Bathymodioline mussels dominate deep-sea methane seep and hydrothermal vent habitats and obtain nutrients and energy primarily through chemosynthetic endosymbiotic bacteria in the bacteriocytes of their gill. However, the molecular mechanisms that orchestrate mussel host-symbiont interactions remain unclear. Here, we constructed a comprehensive cell atlas of the gill in the mussel from the South China Sea methane seeps (1100 m depth) using single-nucleus RNA-sequencing (snRNA-seq) and whole-mount in situ hybridisation.

View Article and Find Full Text PDF

The ultrastructural study on the female reproductive system of the beetle M. brevicauda (Mordellidae) confirmed the positive correlation between the length of the sperm and the size of the female seminal receptacle (Spermatheca). The spermatheca of the species is characterized by an apical bulb-like structure where the spermathecal duct forms numerous folds filled with sperm.

View Article and Find Full Text PDF

Objective: The principal delivery method for CRISPR-based genome editing in insects is now based on microinjection into single cells or embryos. The direct protein transduction systems cannot be employed in aphids because oogenesis occurs without apparent vitellogenesis. Given the limited timing of injection into the embryonic stage in oviparous aphids, a protein delivery system from the hemolymph to the germline and embryos would be a useful tool for genome editing.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!