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Transformation of the Drosophila Sex-Manipulative Endosymbiont Spiroplasma poulsonii and Persisting Hurdles for Functional Genetic Studies
- Source :
- Appl Environ Microbiol
- Publication Year :
- 2020
- Publisher :
- American Society for Microbiology, 2020.
-
Abstract
- Insects are frequently infected by bacterial symbionts that greatly affect their physiology and ecology. Most of these endosymbionts are, however, barely tractable outside their native host, rendering functional genetics studies difficult or impossible. Spiroplasma poulsonii is a facultative bacterial endosymbiont of Drosophila melanogaster that manipulates the reproduction of its host by killing its male progeny at the embryonic stage. S. poulsonii, although a very fastidious bacterium, is closely related to pathogenic Spiroplasma species that are cultivable and genetically modifiable. In this work, we present the transformation of S. poulsonii with a plasmid bearing a fluorescence cassette, leveraging techniques adapted from those used to modify the pathogenic species Spiroplasma citri. We demonstrate the feasibility of S. poulsonii transformation and discuss approaches for mutant selection and fly colonization, which are persisting hurdles that must be overcome to allow functional bacterial genetics studies of this endosymbiont in vivo.<br />IMPORTANCE Dozens of bacterial endosymbiont species have been described and estimated to infect about half of all insect species. However, only a few them are tractable in vitro, which hampers our understanding of the bacterial determinants of the host-symbiont interaction. Developing a transformation method for S. poulsonii is a major step toward genomic engineering of this symbiont, which will foster basic research on endosymbiosis. This could also open the way to practical uses of endosymbiont engineering through paratransgenesis of vector or pest insects.
- Subjects :
- oric
Male
replication
plasmids
animal structures
Spiroplasma
Genetics and Molecular Biology
Paratransgenesis
pure culture
Applied Microbiology and Biotechnology
citri
susceptibility
Bacterial genetics
03 medical and health sciences
stomatognathic system
expression
evolution
Animals
bacteria
Symbiosis
Drosophila
030304 developmental biology
Spiroplasma citri
Genetics
0303 health sciences
endosymbiosis
Ecology
biology
Endosymbiosis
030306 microbiology
Reproduction
fungi
secondary endosymbiont
biochemical phenomena, metabolism, and nutrition
biology.organism_classification
male killing
Transformation (genetics)
Drosophila melanogaster
insect
Female
Transformation, Bacterial
codon
Food Science
Biotechnology
Subjects
Details
- ISSN :
- 10985336 and 00992240
- Volume :
- 86
- Database :
- OpenAIRE
- Journal :
- Applied and Environmental Microbiology
- Accession number :
- edsair.doi.dedup.....1f39c96a35648bb8480e53e1376c09c5
- Full Text :
- https://doi.org/10.1128/aem.00835-20