title: Rapid fluorescence-based screening for Wolbachia endosymbionts in Drosophila germ line and somatic tissues.
abstract: Wolbachia is a globally distributed bacterial endosymbiont present in arthropods and nematodes. The advent of sensitive PCR-based approaches has greatly facilitated the identification of Wolbachia-infected individuals and analysis of population infection levels. Here, a complementary visual fluorescence-based Wolbachia screening approach is described. Through the use of the fluorescent dye Syto-11, Wolbachia can be efficiently detected in various Drosophila tissues, including ovaries. Syto-11 also stains Wolbachia in other insects. Because Wolbachia is inherited through the maternal germ line, bacteria reside in the ovaries of flies in infected populations. An advantage of this staining approach is that it informs about Wolbachia titer as well as its tissue and cellular distribution. Using this method, the infection status of insect populations in two central California locations was determined, and variants with unusually low or high Wolbachia titers were isolated. In addition, a variant with ovarioles containing both infected and uninfected egg chambers was identified. Syto-11 staining of Cardinium- and Spiroplasma-infected insects was also analyzed.
authors.affiliation: Molecular, Cellular, and Developmental Biology, University of California at Santa Cruz, 317 Sinsheimer Laboratories, Santa Cruz, CA 95064, USA. lindley@biology.ucsc.edu
authors.forename: Catharina
authors.initials: C
authors.lastname: Casper-Lindley
authors.forename: Scott
authors.initials: S
authors.lastname: Kimura
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Saxton
authors.forename: Yonathan
authors.initials: Y
authors.lastname: Essaw
authors.forename: Isaac
authors.initials: I
authors.lastname: Simpson
authors.forename: Vinson
authors.initials: V
authors.lastname: Tan
authors.forename: William
authors.initials: W
authors.lastname: Sullivan
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis; D004331:Drosophila melanogaster / Q000382:microbiology*; D005854:Germ Cells / Q000382:microbiology; D008856:Microscopy, Fluorescence / Q000379:methods*; D009696:Nucleic Acids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D013149:Spiroplasma / Q000235:genetics / Q000302:isolation & purification; D013559:Symbiosis; D013997:Time Factors; D020577:Wolbachia / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D009696:Nucleic Acids