Abstract
BackgroundNosema bombycis is a unicellular eukaryotic pathogen of the silkworm, Bombyx mori, and is an economic and occupational hazard in the silkworm industry. Because of its long incubation period and horizontal and vertical transmission, it is subject to quarantine measures in sericulture production. The microsporidian life-cycle includes a dormant extracellular phase and intracellular proliferation phase, with the proliferation period being the most active period. This latter period lacks spore wall protection and may be the most susceptible stage for control.MethodsIn order to find suitable target for the selective breeding of N. bombycis-resistant silkworm strains, we screen highly expressed membrane proteins from the transcriptome data of N. bombycis. The subcellular localization of the candidate protein was verified by Indirect immunofluorescence analysis (IFA) and immunoelectron microscopy (IEM), and its role in N. bombycis proliferation was verified by RNAi.ResultsThe N. bombycis protein (NBO_76g0014) was identified as a transmembrane protein and named NbTMP1. It is homologous with hypothetical proteins NGRA_1734 from Nosema granulosis. NbTMP1 has a transmembrane region of 23 amino acids at the N-terminus. Indirect immunofluorescence analysis (IFA) results suggest that NbTMP1 is secreted on the plasma membrane as the spores develop. Western blot and qRT-PCR analysis showed that NbTMP1 was expressed in all developmental stages of N. bombycis in infected cells and in the silkworm midgut. Downregulation of NbTMP1 expression resulted in significant inhibition of N. bombycis proliferation.ConclusionsWe confirmed that NbTMP1 is a membrane protein of N. bombycis. Reduction of the transcription level of NbTMP1 significantly inhibited N. bombycis proliferation, and this protein may be a target for the selective breeding of N. bombycis-resistant silkworm strains.Graphical
Highlights
Microsporidia are obligate intracellular parasitic eukaryotes with a wide range of hosts, infecting almost all vertebrates and invertebrates, including humans [1,2,3]
The polar tube is ejected when microsporidia are stimulated by suitable conditions, and the infective sporoplasm is transferred into the host cells through the polar tube prior to proliferation [13]
In order to facilitate the control of N. bombycis in the cell, we attempted to screen highly expressed membrane proteins from the transcriptome data of N. bombycis
Summary
Microsporidia are obligate intracellular parasitic eukaryotes with a wide range of hosts, infecting almost all vertebrates and invertebrates, including humans [1,2,3]. These spore-forming unicellular parasites have a unique ultrastructure and life-cycle [3]. The microsporidian life-cycle includes a dormant extracellular phase and intracellular proliferation phase, with the proliferation period being the most active period. This latter period lacks spore wall protection and may be the most susceptible stage for control
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