Abstract
Functional gene analysis tools in Caenorhabditis elegans are often ineffective in other nematodes due to differences in gonadal morphology and transgene silencing. Here, we established a method to generate stable transgenic lines in the nematodes Auanema freiburgense and Tokorhabditis tufae using microparticle bombardment coupled with hygromycin B selection. Despite using non-codon-optimized GFP, transgenic strains expressing fluorescent markers were obtained in both species. Additionally, an Auanema codon-optimized RFP construct showed robust expression in all tissues. This method will be valuable for future studies into the unusual sex determination, viviparity, and stress resistance in Auanema and Tokorhabditis .