{Reference Type}: Journal Article

{Author}: Jun Qian;Haibin Xu;Jingyuan Song;Jiang Xu;Yingjie Zhu;Shilin Chen

{Year}: 2013

{Title}:Genome-wide analysis of simple sequence repeats in the model medicinal mushroom Ganoderma lucidum

{Tag}: 0

{Star}: 0

{Volume}: 512

{Issue}: 2

{Pages}: 331-336

{DOL}: http://dx.doi.org/10.1016/j.gene.2012.09.127

{ISBN/ISSN}:0378-1119

{Keywords}: Agaricomycetes;Simple sequence repeats;SSRs;Polyporales

{Abstract}: Simple sequence repeats (SSRs) or microsatellites are one of the most popular sources of genetic markers and play a significant role in gene function and genome organization. We identified SSRs in the genome of Ganoderma lucidum and analyzed their frequency and distribution in different genomic regions. We also compared the SSRs in G. lucidum with six other Agaricomycetes genomes: Coprinopsis cinerea, Laccaria bicolor, Phanerochaete chrysosporium, Postia placenta, Schizophyllum commune and Serpula lacrymans. Based on our search criteria, the total number of SSRs found ranged from 1206 to 6104 and covered from 0.04% to 0.15% of the fungal genomes. The SSR abundance was not correlated with the genome size, and mono- to tri-nucleotide repeats outnumbered other SSR categories in all of the species examined. In G. lucidum, a repertoire of 2674 SSRs was detected, with mono-nucleotides being the most abundant. SSRs were found in all genomic regions and were more abundant in non-coding regions than coding regions. The highest SSR relative abundance was found in introns (108 SSRs/Mb), followed by intergenic regions (84 SSRs/Mb). A total of 684 SSRs were found in the protein-coding sequences (CDSs) of 588 gene models, with 81.4% of them being tri- or hexa-nucleotides. After scanning for InterPro domains, 280 of these genes were successfully annotated, and 215 of them could be assigned to Gene Ontology (GO) terms. SSRs were also identified in 28 bioactive compound synthesis-related gene models, including one 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), three polysaccharide biosynthesis genes and 24 cytochrome P450 monooxygenases (CYPs). Primers were designed for the identified SSR loci, providing the basis for the future development of SSR markers of this medicinal fungus.

{Author Address}:   http://www.sciencedirect.com

{Language}: English

{Reference Type}: Journal Article

{Author}: Kanako Hatoh;Kosuke Izumitsu;Atsushi Morita;Kiminori Shimizu;Akira Ohta;Masataka Kawai;Takashi Yamanaka;Hitoshi Neda;Yuko Ota;Chihiro Tanaka

{Year}: 2013

{Title}:Transformation of the mushroom species Hypsizigus marmoreus, Flammulina velutipes, and Grifola frondosa by an Agrobacterium-mediated method using a universal transformation plasmid

{Tag}: 0

{Star}: 0

{Volume}: 54

{Issue}: 1

{Pages}: 8-12

{DOL}: http://dx.doi.org/10.1016/j.myc.2012.08.002

{ISBN/ISSN}:1340-3540

{Keywords}: Agrobacterium tumefaciens;Cryptococcus promoter;Maitake;Transgene;Winter mushrooms

{Abstract}: Agrobacterium tumefaciens-mediated transformation (AMT) was successfully applied to mycelia of the 3 economically important mushrooms Hypsizigus marmoreus, Flammulina velutipes, and Grifola frondosa. We used the hygromycin B resistance gene (hph) under the control of the Cryptococcus neoformans actin promoter. Eighty-six resistant strains of H.  marmoreus, 4 of F. velutipes, and 2 of G. frondosa were obtained. All transformants were highly resistant to hygromycin B, suggesting that the C. neoformans actin promoter has a potential universal promoter activity in basidiomycetes. Southern analysis revealed random but single integration of the hph gene.

{Author Address}:   http://www.sciencedirect.com

{Language}: English

{Reference Type}: Journal Article

{Author}: Kaori (Otsuka) Saito;Shinji Kurose;Yoshio Tsujino;Toshiyuki Osakai;Kunishige Kataoka;Takeshi Sakurai;Eiichi Tamiya

{Year}: 2013

{Title}:Electrochemical characterization of a unique, “neutral” laccase from Flammulina  velutipes

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{Star}: 0

{Volume}: 115

{Issue}: 2

{Pages}: 159-167

{DOL}: http://dx.doi.org/10.1016/j.jbiosc.2012.09.011

{ISBN/ISSN}:1389-1723

{Keywords}: Laccase;Flammulina velutipes;Heterologous expression;Direct electron transfer;Redox potential;Optimum pH

{Abstract}: The flac1 gene consisted of 1488 bases encodes a novel laccase (Flac1) from Flammulina velutipes. The deduced amino acid sequence of Flac1 with 496 amino acids shows 58–64% homologies with other fungal laccases. The recombinant Flac1 (rFlac1) was heterologously expressed in Pichia pastoris, with sugars of approximately 4  kDa attached on the protein molecule, which has the calculated molecular mass of 53,532  Da. rFlac1 was shown to be a multi-copper oxidase from spectroscopies. The optimum pHs of rFlac1 for oxidations of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), p-phenylenediamine, and o-aminophenol, were 5.0, 5.0, and 6.0–6.5, respectively, showing higher pH values than those from many other fungal laccases. The slightly acidic or neutral optimum pH that is not strongly dependent on substrates is a unique property of rFlac1. Effective O2 reduction was realized by the direct electron transfer of rFlac1 at a highly oriented pyrolytic graphite electrode modified with fine carbon particles (Ketjen Black) in O2-saturated solution. The pHs showing the maximum ΔE°′ [= E°′(enzyme)  −  E°′(substrate)] coincided well with the optimum pHs shown by rFlac1 under steady-state conditions. The present electrochemical results of rFlac1 indicate that ΔE°′ is one of the primary factors to determine the activity of multi-copper oxidases.

{Author Address}:   http://www.sciencedirect.com

{Language}: English