title: Cloning and functional analysis of a laccase gene during fruiting body formation in Hypsizygus marmoreus.
abstract: The Hypsizygus marmoreus laccase gene (lcc1) sequence was cloned and analyzed. The genomic DNA of lcc1 is 2336 bp, comprising 13 introns and 14 exons. The 1626-bp full-length cDNA encodes a mature laccase protein containing 542 amino acids, with a 21-amino acid signal peptide. Phylogenetic analysis showed that the lcc1 amino acid sequence is homologous to basidiomycete laccases and shares the highest similarity with Flammulina velutipes laccase. A 2021-bp promoter sequence containing a TATA box, CAAT box, and several putative cis-acting elements was also identified. To study the function of lcc1, we first overexpressed lcc1 in H. marmoreus and found that the transgenic fungus producing recombinant laccase displayed faster mycelial growth than the wild-type (wt) strain. Additionally, primordium initiation was induced 3-5 days earlier in the transgenic fungus, and fruiting body maturation was also promoted approximately five days earlier than in the wt strain. Furthermore, we detected that lcc1 was sustainably overexpressed and that laccase activity was also higher in the transgenic strains compared with the wt strain during development in H. marmoreus. These results indicate that the H. marmoreus lcc1 gene is involved in mycelial growth and fruiting body initiation by increasing laccase activity.
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China.
authors.forename: Jinjing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China.
authors.forename: Mingjie
authors.initials: M
authors.lastname: Chen
authors.affiliation: College of Life Science, Nanjing Agricultural University, No. 1, Weigang road, XuanWu District, Nanjing 210095, China.
authors.forename: Ang
authors.initials: A
authors.lastname: Ren
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China.
authors.forename: Jianchun
authors.initials: J
authors.lastname: Huang
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Life Science, Nanjing Agricultural University, No. 1, Weigang road, XuanWu District, Nanjing 210095, China.
authors.forename: Mingwen
authors.initials: M
authors.lastname: Zhao
authors.affiliation: National Research Center for Edible Fungi Biotechnology and Engineering, Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture, the People's Republic of China, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, No. 1000, Jinqi Road, FengXian District, Shanghai 201403, China; College of Life Science, Nanjing Agricultural University, No. 1, Weigang road, XuanWu District, Nanjing 210095, China. Electronic address: feng_zy@yahoo.com.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Feng
mesh_terms: D000363:Agaricales / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics; D003001:Cloning, Molecular; D004271:DNA, Fungal; D055393:Flammulina / Q000235:genetics; D048690:Fruiting Bodies, Fungal; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D042845:Laccase / Q000235:genetics*; D030781:Organisms, Genetically Modified; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment
chemical_list: D004271:DNA, Fungal; D042845:Laccase
keywords: Fruiting body; Hypsizygus marmoreus; Laccase; Overexpression; Promoter