Identification of a microsporidian isolate from Cnaphalocrocis Medinalis and its pathogenicity to Bombyx mori

A microsporidian, CmM2, was isolated from Cnaphalocrocis medinalis. The biological characters, molecular analysis and pathogenicity of CmM2 were studied. The spore of CmM2 is long oval in shape and 3.45 ± 0.25 × 1.68 ± 0.18 µm in size, the life cycle includes meronts, sporonts, sporoblasts, and spores, with typical diplokaryon in each stage, propagated in binary fission. There is positive coagulation reaction between CmM2 and the polyclonal antibody of Nosema bombycis (N.b.). CmM2 spores is binuclear, and has 10–12 polar filament coils. The small subunit ribosomal RNA (SSU rRNA) gene sequence of CmM2 was obtained by PCR amplification and sequencing, the phylogenetic tree based on SSU rRNA sequences had been constructed, and the similarity and genetic distance of SSU rRNA sequences were analyzed, showed that CmM2 was grouped in the Nosema clade. The 50% infectious concentration of CmM2 to Bombyx mori is 4.72 × 104 spores ml−1, and the germinative infection rate is 12.33%. The results showed that CmM2 is classified into genus Nosema, as Nosema sp. CmM2, and has a heavy infectivity to B. mori. The result indicated as well that it is valuable taxonomic determination for microsporidian isolates based on both biological characters and molecular evidence.

[1]  Xing Dong Biological Characters and Molecular Phylogenesis of a Microsporidium Isolated from Delias pasithoe(Lepidoptera:Pieridae) , 2011 .

[2]  R. Bastrop,et al.  Molecular characterisation of the Microsporidia of the amphipod Gammarus duebeni across its natural range revealed hidden diversity, wide-ranging prevalence and potential for co-evolution. , 2010, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[3]  Luo Shoujin Occurrence of rice leaf roller in China and its identification and prevention. , 2010 .

[4]  Li Xu,et al.  Sequence and Phylogenetic Analysis of SSU rRNA Gene of Five Microsporidia , 2009, Current Microbiology.

[5]  Yongji Wan,et al.  [Cloning and phylogenetic analysis of small subunit ribosomal RNA core sequence of Vairimorpha ceraces (Microspora: Burenellidae) from the insect of Lepidoptera, Cerace Stipatana (Walker)]. , 2008, Wei sheng wu xue bao = Acta microbiologica Sinica.

[6]  L. Fu Characterization of a Microsporidium Isolated From Larvae of Beet Armyworm,Laphygma exigua H. , 2007 .

[7]  W. T. Tay,et al.  Complete rRNA Gene Sequences Reveal that the Microsporidium Nosema bombi Infects Diverse Bumblebee (Bombus spp.) Hosts and Contains Multiple Polymorphic Sites , 2005, The Journal of eukaryotic microbiology.

[8]  J. Larsson Molecular versus morphological approach to microsporidian classification. , 2005, Folia parasitologica.

[9]  Chung-Hsiung Wang,et al.  Complete Sequence and Gene Organization of the Nosema spodopterae rRNA Gene , 2005, The Journal of eukaryotic microbiology.

[10]  W. Yong A NEW MICROSPORIDIUM(MICROSPOREDIA,BURENELLIDAE) FROM THE INSECT OF LEPIDOPTERA, CERACE STIPATANA (WALKER) , 2005 .

[11]  P. Brindley,et al.  Epidemiology of microsporidiosis: sources and modes of transmission. , 2004, Veterinary parasitology.

[12]  S. Kanginakudru,et al.  Phylogenetic relationships of three new microsporidian isolates from the silkworm, Bombyx mori. , 2004, Journal of invertebrate pathology.

[13]  C. Lo,et al.  The novel organization and complete sequence of the ribosomal RNA gene of Nosema bombycis. , 2004, Fungal genetics and biology : FG & B.

[14]  N. Simões,et al.  A new isolate of the microsporidium Vairimorpha necatrix (Microsporidia, Burenellidae) recorded in the azores. , 2004, Journal of invertebrate pathology.

[15]  Cao Yang,et al.  Studies on the Application of PCR Molecular Diagnosis to Silkworms with Simulated Pebrine Disease , 2004 .

[16]  T. Kawarabata Biology of Microsporidians Infecting the Silkworm, Bombyx mori, in Japan. , 2003 .

[17]  Fang Ding A microsporidium (Mic-I) isolated from Mimastra cyanura Hope , 2002 .

[18]  P. Keeling,et al.  Microsporidia: biology and evolution of highly reduced intracellular parasites. , 2002, Annual review of microbiology.

[19]  P. Adler,et al.  Molecular characterization and taxonomy of a new species of Caudosporidae (Microsporidia) from black flies (diptera: simuliidae) with host-derived relationships of the North American caudosporids. , 2000, Journal of invertebrate pathology.

[20]  E. Canning,et al.  Vairimorpha imperfecta n.sp., a microsporidian exhibiting an abortive octosporous sporogony in Plutella xylostella L. (Lepidoptera: Yponomeutidae) , 1999, Parasitology.

[21]  M. Wittner Historic Perspective on the Microsporidia: Expanding Horizons , 1999 .

[22]  C. Vossbrinck,et al.  Phylogenetic relationships among Vairimorpha and Nosema species (Microspora) based on ribosomal RNA sequence data. , 1994, Journal of invertebrate pathology.

[23]  E. I. Hazard,et al.  Taxonomy of phylum microspora. , 1992, Critical reviews in microbiology.

[24]  R. Ishihara Dimorphic development of Nosema bombycis spores in gut epithelium of larvae of the silkworm, Bombyx mori , 1991 .

[25]  Y. Abe Nosema mesnili isolated from the fall webworm, Hyphantria cunea , 1988 .

[26]  J. Larsson Identification of Microsporidian Genera (Protozoa, Microspora) - a Guide with Comments on the Taxonomy , 1988 .

[27]  I. Fries,et al.  Nosema Apis, Sampling Techniques and Honey Yield , 1984 .

[28]  V. Sprague Classification and Phylogeny of the Microsporidia , 1977 .