title: A simple and efficient genetic transformation method of Ganoderma weberianum.
abstract: In this study, the Agrobacterium tumefaciens-mediated transformation method for Ganderma weberianum has been established. Driven by the cauliflower mosaic virus (CaMV) 35S promoter, the hygromycin phosphotransferase (hpt), β-glucuronidase (uidA), and enhanced green fluorescent protein (egfp) genes have been efficiently expressed in transgenic mycelia and spores. The transformation system was composed of the growing mycelia, A. tumefaciens strain GV3101, and the expression vector pBI-H1, harboring the CaMV 35S promoter and selective hpt marker. The genetic transformation of G. weberianum was achieved through co-cultivation of Agrobacterium lawn and fungal mycelia at 28 °C on yeast extract agar (YEA) medium. Stable genetic transformants were obtained through successive hygromycin B selections and single spore isolation. Over 80 % of transformants showed genetic stability even after ten rounds of subculturing. The simple and efficient genetic transformation method is a useful tool for molecular genetics analyses and gene manipulation of G. weberianum.
authors: Guangzhou Key Laboratory for Functional Study on Stress-Resistant Genes in Plants, School of Life Sciences, Guangzhou University, Guangzhou, 510006, China.
authors: Yu-Ping
authors: YP
authors: Zhou
authors: Min-Hua
authors: MH
authors: Chen
authors: Jun-Jie
authors: JJ
authors: Lu
authors: Xun
authors: X
authors: Kang
authors: Qiong-Hua
authors: QH
authors: Chen
authors: Xiao-Ling
authors: XL
authors: Huang
authors: Chang-En
authors: CE
authors: Tian
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000378:metabolism; D032902:Ganoderma / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D014170:Transformation, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
references: Biotechnol Adv. 2013 Dec;31(8):1562-74
references: 23988676
references: Curr Genet. 2005 Jul;48(1):1-17
references: 15889258
references: Acta Pharmacol Sin. 2004 Jun;25(6):833-8
references: 15169641
references: Plant Cell Physiol. 1993 Mar;34(2):345-55
references: 8199776
references: Bioresour Technol. 2008 Nov;99(17):8524-7
references: 18434137
references: Nat Commun. 2012 Jun 26;3:913
references: 22735441
references: Mol Biotechnol. 2002 Mar;20(3):239-44
references: 11936254
references: Nat Biotechnol. 1998 Sep;16(9):839-42
references: 9743116
references: EMBO J. 1995 Jul 3;14(13):3206-14
references: 7621833
references: Microbiol Res. 2011 May 20;166(4):314-22
references: 20869218
references: Folia Microbiol (Praha). 2004;49(2):112-6
references: 15227780
references: BMC Biotechnol. 2010 Sep 14;10 :67
references: 20836896
references: World J Microbiol Biotechnol. 2012 Jan;28(1):283-91
references: 22806804
references: Mycol Res. 2009 Mar;113(Pt 3):290-7
references: 19059480
references: Plant Biotechnol J. 2007 Mar;5(2):221-9
references: 17309677
references: Peptides. 2006 Jan;27(1):27-30
references: 16039755
references: Biosci Biotechnol Biochem. 2010;74(11):2327-9
references: 21071861
references: FEMS Microbiol Lett. 2004 Apr 15;233(2):201-4
references: 15063487
references: Biotechnol Appl Biochem. 2006 Mar;43(Pt 3):181-6
references: 16302847
references: J Microbiol Methods. 2013 Nov;95(2):251-2
references: 24050950