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Two novel strains of Wolbachia coexisting in both species of mulberry leafhoppers
1Department of Insect Genetics and Evolution, National Institute of Agrobiological Sciences, Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
Abstract:
Wolbachia is an intracellular symbiont that causes reproductive disorders in many insects. Its presence in the leafhoppers Hishimonoides sellatiformis and Hishimonus sellatus, vectors of mulberry dwarf-Phytoplasma, was confirmed by the PCR analysis of 16S rDNA, ftsZ and wsp. Sequencing of cloned PCR products revealed that two Wolbachia strains coexist in both leafhoppers. The phylogenetic analysis of wsp revealed that these strains belong in novel positions in the B-group of Wolbachia. These strains were detected by PCR and/or PCR-RFLP in all of the tested non-genital organs including salivary glands, as well as in the tested genital organs of Hishimonoides sellatiformis. In addition, Wolbachia-like organisms were observed by electron microscopy in all PCR-positive organs. We discuss the possible horizontal transmission of Wolbachia via mulberry trees.
Insights
Two novel Wolbachia strains were found in mulberry leafhoppers, insect vectors of dwarf-Phytoplasma. These bacteria were detected in all tested organs, suggesting potential horizontal transmission via mulberry trees.
Area of Science:
- Entomology
- Microbiology
- Molecular Biology
Background:
- Wolbachia are intracellular symbionts impacting insect reproduction.
- Leafhoppers Hishimonoides sellatiformis and Hishimonus sellatus transmit mulberry dwarf-Phytoplasma.
- Understanding Wolbachia in these vectors is crucial for disease management.
Purpose of the Study:
- To confirm the presence and characterize Wolbachia strains in H. sellatiformis and H. sellatus.
- To investigate the distribution of Wolbachia within insect organs.
- To explore potential transmission routes of Wolbachia.
Main Methods:
- Polymerase Chain Reaction (PCR) using 16S rDNA, ftsZ, and wsp genes.
- Sequencing of PCR products for strain identification.
- Phylogenetic analysis of wsp gene sequences.
- PCR-Restriction Fragment Length Polymorphism (PCR-RFLP).
- Electron microscopy for visualizing Wolbachia-like organisms.
Main Results:
- Wolbachia presence confirmed in both leafhopper species using PCR.
- Sequencing revealed two distinct Wolbachia strains in each species.
- Phylogenetic analysis placed these strains in novel positions within the B-group.
- Wolbachia were detected in all tested genital and non-genital organs, including salivary glands.
- Electron microscopy confirmed Wolbachia-like organisms in PCR-positive organs.
Conclusions:
- H. sellatiformis and H. sellatus harbor at least two novel Wolbachia strains.
- Wolbachia exhibit a widespread distribution within these insect vectors.
- Horizontal transmission of Wolbachia through mulberry trees is a plausible route.
