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• W45388230 abstract "The bacterium Moraxella osloensis associated with the nematode Phasmarhabditis hermaphrodita is a pathogen of slug Deroceras reticulatum. Nematode infective juveniles vector M. osloensis into the shell cavity of D. reticulatum, and the slug is killed within 4-16 days. Gene expression profile of M. osloensis in the slug was analyzed and differentially expressed genes were identified using a selective capture of transcribed sequences (SCOTS) technique. Most identified sequences were homologous to other bacteria, and function as putative cell structure, energy metabolism, degradation, translocation, and unknown function proteins. Other identified sequences did not exhibit similarity to any genes or gene products in current databases, and are thus novel. We further evaluated the role of SCOTS identified two genes, M5 (protein-disulfide isomerase) and M8 (protein kinase), in the virulence of M. osloensis to the slug using mutation and complementation strategy. Mutants were constructed with insertion-deletion strategy by inverse PCR. The full length of M5 and M8 fragments were obtained by inverse PCR to complement mutants. Compared to the wild type, the virulence of M. osloensiss mutants to the slug was markedly reduced and could be complemented. Therefore, we conclude that the in vivo expressed genes, M5 and M8, contribute to M. osloensis virulence to the slug. This study provided a first glimpse to the gene expression in a mollusk host during bacterial infection, and insights into understanding the mechanism of the bacterial virulence to a mollusk host . Grey garden slugs, Deroceras reticulatum, are regarded as the most serious pests of a wide range of agricultural and horticultural plants (Fig-1). The nematode Phasmarhabditis hermaphrodita, associated with bacterium Moraxella osloensis has potential for biological control of D. reticulatum. P. hermaphrodita vectors bacterium M. osloensis into the shell cavity of D. reticulatum, and the bacterium is the main killing agent. Genes expressed by M. osloensis in slugs are likely to play important roles in virulence. Identification of these genes can help in designing improved bacterial strains. The objective of this study was to identify the virulence genes expressed by M. osloensis in slugs after infection. Selective capture of transcribed sequences (SCOTS) analysis was employed to identify genes expressed by M. osloensis in the slug D. reticulatum at four days post infection. The roles of SCOTS captured genes were analyzed and evaluated based on the Genomic database and mutagenesis studies. Selective Capture of Transcribed Sequences Preparation of Nucleic Acids: The M. osloensis genomic DNA was prepared. Total RNA was isolated from four days post-infected slugs (in-vivo) and from broth cultured midlog phase bacteria (in-vitro). The isolated RNA was converted to сDNA by using random primers with an adapter overhang. Bacterial Inoculation Slug D. reticulatum was collected from the field and fed on fresh carrots for at least 12 days. Only healthy adult slugs were used in the experiments. The Pure culture of M. osloensis was introduced into the shell cavity of slug D. reticulatum via injection. Ruisheng An1, Srinand Sreevatsan2, and Parwinder Grewal1 1. Department of Entomology, the Ohio State University, OARDC, Wooster, OH 44691, USA 2. Veterinary Population Medicine Department, College of Veterinary Medicine, St. Paul, MN 55108 INTRODUCTION MATERIALS & METHODS The overall hypothesis is that M. osloensis will express genes exclusively in slugs that will help M. osloensis to survive, replicate and kill the slug. HYPOTHESIS Fig-1: The left, D. reticulatum feeding on oil seed rape seedlings. The right, healthy D. reticulatum on left and infected D. reticulatum on right with swollen mantle. Bacterial cultures grown within slug host Isolate total RNA and convert to linkered cDNA" @default.
• W45388230 created "2016-06-24" @default.
• W45388230 creator A5057365183 @default.
• W45388230 date "2006-04-17" @default.
• W45388230 modified "2023-09-27" @default.
• W45388230 title "Identification of the first two virulence genes of Moraxella osloensis in the slug Deroceras reticulatum" @default.
• W45388230 cites W2115335843 @default.
• W45388230 hasPublicationYear "2006" @default.
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