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Do microsporidia function as ＂biological weapon＂ for Harmonia axyridis under natural conditions？

Insect Science 2015年第3期22卷 353-359页

作者： Tobias Gegner Oliver Otti Simon Tragust Heike Feldhaar Animal Population Ecology Department of Animal Ecology I Bayreuth Center of Ecology and Environmental Research (BayCEER) University of Bayreuth D-95440 Bayreuth Germany

Invasive alien species, such as the multicoloured Asian ladybird Harmonia axyridis, are often regarded as major drivers ofbiodiversity loss. Therefore understanding which characteristics or mechanisms contribute to their invasive success is important. Here the role of symbiotic microsporidia in the hemolymph of H. axyridis was investigated in the context of intraguild predation between wild-caught H. axyridis and the native ladybird species Coccinella septempunctata. The microsporidia were recently discussed to contribute to the unpalatability of Harmonia for other coccinellids during intraguild predation and to function as ＂biological weapons＂. In the present study, visual detection of microsporidia in hemolymph samples revealed that 73.5 % of H. axyridis were infected. Intraguild predation experiments between larvae of the two species showed a significant competitive advantage for H. axyridis, even against larger larvae of C. septempunctata. Adult C. septempunctata always killed and fed on H. axyridis larvae. However only 11.4 % （4 of 47） of C. septempunctata that fed on infected H. axyridis died within 4 months. In contrast to previous studies this suggests that microsporidia or harmonine, the chemical defense compound of H. axyridis, do not lead to death of C. septempunctata preying on larvae ofH. axyridis. Instead our results support the idea that competitive advantage during intraguild predation greatly facilitates the success of H. axyridis and that this may help this highly invasive species to outcompete native species. The impact of microsporidia on Harmonia itself as well as on interspecific interactions require further studies.

关键词： defensive symbiont endosymbiont Harmonia axyridis invasive species microsporidia

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Evolutionary ecology of microsporidia associated with the invasive ladybird Harmonia axyridis

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Insect Science 2015年第3期22卷 313-324页

作者： Andreas Vilcinskas Henrike Schmidtberg Arnaud Estoup Ashraf Tayeh Benoit Facon Heiko Vogel Institute of Phytopathology and Applied Zoology Justus-Liebig-University of Giessen Heinrich-Buff-Ring 26-32 Giessen D-35392 Germany Fraunhofer Institute for Molecular Biology and Applied Ecology Winchester Strasse 2 Giessen D-35394 Germany UMR 1062 CBGP Inra 34988 Montpellier France Max Planck Institute for Chemical Ecology Hans-Knoll-Strasse 8 D-07745 Jena Germany

Invasive species are characterized by the rapid growth and spread of their populations after establishing a foothold in new habitats, and there are now many examples of such species negatively affecting biodiversity and the economy. It is unclear why some species can become successful invaders, whereas most （even if closely related） remain noninvasive. We previously proposed a hypothesis that parasites associated with invading species can promote their invasive success if they are harmless toward the invaders but harmful to their competitors and/or predators in the newly colonized habitat. Here we discuss whether microsporidia that have recently been discovered in the invasive ladybird Harmonia axyridis contribute to its invasive success. We show that all H. axyridis beetles sourced from diverse collection sites all over the world carry abundant microsporidia. This suggests that both native and invasive H. axyridis populations are associated with these tolerated parasites, which were likely to have existed in native populations before expansion rather than being acquired in newly colonized areas. We describe the pathogenesis of the microsporidia during different developmental stages of H. axyridis and we address the possibility that the predation of its infected eggs and larvae by competing native ladybird species may lead to their infection and ultimately to their decline. Finally, we discuss our initial hypothesis： microsporidia that are tolerated by an invasive vector insect can be active against susceptible native competitors and/or predator species.

关键词： Coccinellidae evolutionary ecology Harmonia aryridis innate immunity intraguild predation invasion biology microsporidia

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Phagocytic Uptake of Nosema bombycis (microsporidia) Spores by Insect Cell Lines

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Journal of Integrative Agriculture 2012年第8期11卷 1321-1326页

作者： CAI Shun-feng LU Xing-meng QIU Hai-hong LI Ming-qian FENG Zhen-zhen Laboratory of Invertebrate Pathology Zhejiang UniversityHangzhou 310058P.R.China

microsporidia are highly specialized obligate intracellular parasites that can infect a wide variety of animals ranging from protists to mammals. The classical concept of the parasite invasion into a host cell involves its polar tube acting as a needle-syringe system. However, recent studies show microsporidian spores can also gain access to host cells by phagocytosis. The present study investigated the phagocytic uptake process of causative agent of the pebrine disease, Nosema bombycis, in several insect cell lines. We observed KOH-treated spores and cold-storaged spores can be easily uptaken by all the studied cell types 4 h post inoculation. In contrast, large numbers of freshly recovered spores remained in the culture medium. To further investigate the intracellular fates of KOH-treated spores and cold-storaged spores, electron and fluorescence microscopy were performed. No intracellular germination or subsequent parasite development were observed. Intracellular spores can be detected in host cells by polyclonal antibody 7 d post inoculation, suggesting phagocytized N. bombycis could not be digested by these non-professional phagocytes. Our results suggest that, phagocytic uptake of N. bombycis spores might represent a defense mechanism of the host cells and the intact spore wall barrier enable freshly recovered spores to keep resistance to this mechanism.

关键词： microsporidia Nosema bombycis phagocytosis cell invasion

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Morphological and molecular studies of Vairimorpha necatrix BM, a newstrain of the microsporidium ***（microsporidia,burenellidae） recorded in the silkworm,Bombyx mori

Morphological and molecular studies of Vairimorpha necatrix ...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者：罗波 Handeng Liu Guoqing Pan Tian Li Zeng Li Xiaoqun Dang Tie Liu 周泽扬 State Key Laboratory of Silkworm Genome Biology Southwest University Experimental Teaching Center Chongqing Medical University Laboratory of Animal Biology Chongqing Normal University

Vairimorpha sp. BM（2012） is a recent isolate of the microsporidia from the silkworm in Shandong, China. The ultrastructure, tissue pathology and molecular characterization of this isolate is described in this study. This pathogenic fungus causes pebrine disease in silkworms which manifests as a systemic infection. Meanwhile, the silkworm eggs produced by the infected moths were examined using a microscope and PCR amplification. Neither spores nor the expected PCR band were observed, suggesting that no vertical transmission occurred in Bombyx mori. In addition, the ultrastructure of the isolate was studied by light microscopy and transmission electron microscopy. Two types of spores were observed: diplokaroytic spores with 13–17 coils of polar tubes and monokaryotic spores with less coils of polar tubes which could form octospores;however, no sporophorous vesicles were observed. Finally, phylogenetic analysis of the small subunit r RNA genes of Vairimorpha species showed that this isolate has a closer relationship to Vairimorpha necatrix than the other species studied. This result also is supported by phylogenetic analysis based on their actin genes, heat shock protein 70（ HSP70） and RNA polymerase Ⅱ（ RPB1）. Based on the information gained during this study, we propose that this microsporidian species infecting B. mori should be given the name V. necatrix BM.

关键词： Silkworm microsporidia V.necatrix Ultrastructure Vertical transmission

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Development of an approach to analyze the interaction between Nosema bombycis（microsporidia） deproteinated chitin spore coats and spore wall proteins

Development of an approach to analyze the interaction betwee...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者：杨东林 Xiaoqun Dang Rui Tian Mengxian Long Chunfeng Li Tian Li Jie Chen Zhi Li Guoqing Pan 周泽扬 State Key Laboratory of Silkworm Genome Biology Southwest University Laboratory of Animal Biology Chongqing Normal University

Nosema bombycis is an obligate intracellular parasite of the Bombyx mori insect. The spore wall of N. bombycis is composed of an electron-dense proteinaceous outer layer and an electron-transparent chitinous inner layer, and the spore wall is connected to the plasma membrane. In this study, the deproteinated chitin spore coats（DCSCs） were acquired by boiling N. bombycis in 1 M Na OH. Under a transmission electron microscope, the chitin spore coat resembles a loosely curled ring with strong refractivity;organelles and nuclei were not observed inside the spore. The anti-SWP25, 26, 30 and 32 antibodies were used to detect whether spore wall proteins within the total soluble and mature spore proteins could bind to the DCSCs. Furthermore, a chitin binding assay showed that within the total soluble and mature spore proteins, the SWP26, SWP30 and SWP32 spore wall proteins, bound to the deproteinated chitin spore coats, although SWP25 was incapable of this interaction. Moreover, after the DCSCs were incubated with the alkali-soluble proteins, the latter were obtained by treating N. bombycis with 0.1 M Na OH. Following this treatment, SWP32 was still capable of binding the DCSCs, while SWP26 and SWP30 were unable to bind. Collectively, the DCSCs are useful for investigating the arrangement of spore wall proteins, and they shed light on how the microsporidia spore wall is self-assembled.

关键词： Nosema bombycis Deproteinated chitin spore coats(DCSCs) Spore wall protein microsporidia

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Characterization of a novel otubain-like protease with deubiquitination activity from Nosema bombycis（microsporidia）

Characterization of a novel otubain-like protease with deubi...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者：王营 Xiaoqun Dang Bo Luo Chun feng Li Mengxian Long Tian Li Zhi Li Guoqing Pan 周泽扬 State Key Laboratory of Silkworm genome Biology Southwest University Laboratory of Animal Biology Chongqing Normal University

Otubains are a recently identified family of deubiquitinating enzymes（DUBs）. They are involved in diverse biological processes including protein degradation, signal transduction, and cell immune response. Several microsporidian genomes have been published in the last decade;however, little is known about the otubain-like protease in these widely-spread obligate intracellular parasites. Here, we characterized a 25 kDa otubain-like protease（NbOTU1） from the microsporidian Nosema bombycis, the pathogen causing pebrine disease in the economically important insect Bombyx mori. Sequence analysis showed that this protein contained a conserved catalytic triad of otubains composed of aspartate, cysteine, and histidine residues. The expression of Nbotu1 began on day 3 postinfection as determined by the RT-PCR method. Immunofluorescence analysis indicated that NbOTU1 is localized on the spore wall of N. bombycis. The subcellular localization of the NbOTU1 was further detected with immunoelectron microscopy, which showed that NbOTU1 is localized at the regions around endospore wall and plasma membrane. Deubiquitination analysis confirmed that the recombinant NbOTU1 possessed deubiquitination activity in vitro. Taken together, a novel microsporidian otubain-like protease NbOTU1 was partially characterized in N. bombycis, demonstrating its subcellular location and deubiquitination activity. This study provided a basic reference for further dissecting the function of otubains in microsporidia.

关键词： microsporidia Deubiquitinating enzymes Otubain-like protease Nosema bombycis

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Mitochondrial pyruvate dehydrogenase E1 of Nosema bombycis:A marker in microsporidian evolution

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Current Zoology 2009年第6期55卷 423-429页

作者： Tian LI Xiaoqun DANG Jinshan XU Handeng LIU Guoqing PAN Zeyang ZHOU The Key Sericultural Laboratory of Agricultural Ministry Southwest University Chongqing 400716 China Laboratory of Animal Biology Chongqing Normal University Chongqing 400047 China

microsporidia are a group of intracelluar eukaryotic parasites, which can infected almost all animals, including human beings. Till now, no mitochodria but mitosome, a remnant of mitochondria was discovered in this phylum. We present here the mitochondrial pyruvate dehydrogenase El （PDH, including PDHα and PDHβ） of the microsporidian Nosema bombycis, the pathogen of silkworm pebrine. Compared with PDH of microsporidian Encephalitozoon cuniculi and Antonospora locustae, both subunits are eonscrced. The phylogeny indicated that both subunits are mitochondrial. The syntenic maps revealed the subunits organization of NbPDH is distributed in different scaffolds, similar to that of EcPDH but different with AIPDH, and the relationship between phylogeny tree and organization of PDH suggest that the AlPDH subunits organization is the ancestral style of microsporidia, and through the genome evolution, the reshuffling of the chromosome of microsporidia occurred, the adjacent style of ALPDHE1 organization changed, and the two subunits separated and located to different chromosomes in E. cuniculi. For N. bombycis and N. ceranae, they locate to different scaffolds. In order to determine NbPDH subcellular localizations, we prepared the polyclonal antibodies against NbPDH prokaryotic fusion proteins, and adopted the colloidal gold immunological electron microscopy, the expression signals of NbPDH were observed in spores however, the subcellular localization were not definited. In general, through comparison of three mierosporidian PDH molecular phylogeny, subunits organization in chromosomes, localization indicated that PDH is an interesting marker in microsporidia evolution

关键词： microsporidia Nosema bombycis PDH Synteny Immunoloealization Evolution

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Anti-microsporidial effect of thymoquinone on Encephalitozoon intestinalis infection in vitro

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Asian Pacific Journal of Tropical Biomedicine 2020年第1期10卷 42-46页

作者： Ulfet Cetinkaya Gülay Sezer Arzuv Charyyeva Halil Bayraktar Health Vocational College Erciyes UniversityKayseriTurkey School of Medicine Department of PharmacologyErciyes UniversityKayseriTurkey Genkok Genome and Stem Cell Centre Department of Molecular Biology and GeneticsErciyes UniversityKayseriTurkey Life Science Research Centre Faculty of ScienceOstrava UniversityOstravaCzech Republic

Objective:To evaluate the anti-microsporidial effects of the active component of Nigella sativa seeds,thymoquinone,against Encephalitozoon intestinalis using an in vitro ***:Anti-microsporidial effect of thymoquinone against Encephalitozoon intestinalis was evaluated by using various concentrations of thymoquinone(0,1,5,10,15,20,30,35,and 40μM)and sterile dimethyl *** time PCR was used to evaluate the inhibitory effects of thymoquinone on the life cycle of Encephalitozoon ***:The cytotoxic effect of thymoquinone on HEK293 cell line was observed with 30,35,and 40μM concentrations of thymoquinone after 24,48,and 72 hours of *** was observed that 10,15,20,and 30μM concentrations of thymoquinone decreased the spore density compared with the control;however,it was significant only at 30μ***:Thymoquinone shows potent anti-microsporidial effects against Encephalitozoon intestinalis in the in vitro model;however,the toxic concentrations of thymoquinone are also toxic to the host cells.

关键词： microsporidia Thymoquinone Nigella sativa Antiparasitic Encephalitozoon intestinalis

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Natural selection maintains the transcribed LTR retrotransposons in Nosema bombycis

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Journal of Genetics and Genomics 2010年第5期37卷 305-314页

作者： Heng Xiang Guoqing Pan Ruizhi Zhang Jinshan Xu Tian Li Wenle Li Zeyang Zhou Zhonghuai Xiang Institute of Sericulture and Systems Biology Southwest University Beibei Chongqing 400715 China College of Life Science Chongqing Normal University Chongqing 400047 China Biotechnology Research Center Southwest University Beibei Chongqing 400715 China

Eight intact LTR retrotransposons （Nbr1-Nbr8） have been previously characterized from the genome of Nosema bombycis, a eu- karyotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements （Nbr9-Nbr14） identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions （Nbr12）, deletions （Nbrll） and in-frame stop codons in coding regions （Nbr9） were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bornbycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbrll is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbrll retrotransposon. Unlike other transposable elements, Nbrll has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection.

关键词： microsporidia LTR retrotransposon transcribed purifying selection positive selection

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Nitric oxide mediates midgut immunity under microsporidian infection in Antheraea pernyi

Nitric oxide mediates midgut immunity under microsporidian i...

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中国蚕学会2021年学术年会

作者： Luo Yutong Liu Wei Shi Shenglin 王勇 College of Bioscience and Biotechnology Shenyang Agricultural University

The insect gut participates in initial local immune responses by producing reactive oxygen and nitrogen species as well as anti-microbial peptides to resist pathogenic *** oxide（NO）,a signaling and an immune effector molecule synthesized by the enzyme NO synthase（NOS）,mediates an early step of the signal transduction *** this study,we evaluated NO levels after Nosema pernyi infection in Antheraea pernyi *** activity was higher in the microsporidia-infected gut of *** than in that of *** NOS-related genes were cloned,and their spatio-temporal expression patterns were ***2 was expressed quickly in the midgut after *** *** nitroprusside,dihydrate（SNP）,or Nω-L-nitro-arginine methyl ester,hydrochloride（L-NAME）,altered the NO content in *** ***-microbial peptides（AMPs） in the groups exposed to *** plus SNP and *** plus L-NAME exhibited higher and lower expression,respectively,relative to the *** results indicate that microsporidia infection triggers short term activation of NO and NOS genes in the *** gut that is downregulated after 24 ***,infection rates can be influenced by a NOS ***,NO can be induced by ***,NO content in the *** gut also influences AMPs in humoral immunity and some immune-related *** results suggest that nitric oxide plays a vital role in *** gut immunity.

关键词： Antheraea pernyi microsporidia Nitric oxide Antimicrobial peptide

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