title: Development of a transformation system for Hirsutella spp. and visualization of the mode of nematode infection by GFP-labeled H. minnesotensis.
abstract: Hirsutella rhossiliensis and H. minnesotensis are endoparasitic fungi of the second-stage juvenile (J2) of the soybean cyst nematode (Heterodera glycines) in nature. They also parasitize both H. glycines J2 and Caenorhabditis elegans on agar plates. Agrobacterium tumefaciens-mediated transformation conditions were established for these Hirsutella spp. The resulting transformants were similar to the corresponding wild-type strains. The infection processes of H. glycines J2 and C. elegans second larval stage (L2) by H. minnesotensis expressing ZsGreen were microscopically analyzed. Conidia of H. minnesotensis adhered to passing nematodes within 8 h post-inoculation (hpi), formed an infection peg between 8 and 12 hpi, and penetrated the nematode cuticle between 12 and 24 hpi for C. elegans L2 and between 12 and 32 hpi for H. glycines J2. Hyphal proliferation inside of the nematode coelom was observed at approximately 32 hpi for C. elegans L2 and at approximately 40 hpi for H. glycines J2. The fungus consumed the whole body and grew out to produce conidia at approximately 156 and 204 hpi for C. elegans L2 and H. glycines J2, respectively. The efficient transformation protocol and a better understanding of infection process provide a solid foundation for studying the molecular and cellular mechanisms underlying fungal parasitism of nematodes.
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, No. 3 Park 1, West Beichen Road, Chaoyang District, Beijing 100101, China.
authors.forename: Jingzu
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Plant Pathology and Environmental Microbiology, Pennsylvania State University, University Park, PA 16802, USA.
authors.forename: Sook-Young
authors.initials: SY
authors.lastname: Park
authors.affiliation: Department of Plant Pathology and Environmental Microbiology, Pennsylvania State University, University Park, PA 16802, USA.
authors.forename: Seogchan
authors.initials: S
authors.lastname: Kang
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, No. 3 Park 1, West Beichen Road, Chaoyang District, Beijing 100101, China.
authors.forename: Xingzhong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Life Science, Fujian Agriculture and Forestry University, No. 15 Shangxiadian Road, Cangshan District, Fuzhou 350002, China.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Qiu
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, No. 3 Park 1, West Beichen Road, Chaoyang District, Beijing 100101, China.
authors.forename: Meichun
authors.initials: M
authors.lastname: Xiang
mesh_terms: D000818:Animals; D001203:Ascomycota / Q000502:physiology*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics*; D009348:Nematoda / Q000382:microbiology*; D013172:Spores, Fungal; D014170:Transformation, Genetic
chemical_list: D049452:Green Fluorescent Proteins