» Articles » PMID: 25655754

A Simple and Efficient Genetic Transformation Method of Ganoderma Weberianum

Overview
Publisher Springer
Specialty Microbiology
Date 2015 Feb 7
PMID 25655754
Authors
Affiliations
Soon will be listed here.
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.

References
1.
Jiang D, Zhu W, Wang Y, Sun C, Zhang K, Yang J . Molecular tools for functional genomics in filamentous fungi: recent advances and new strategies. Biotechnol Adv. 2013; 31(8):1562-74. DOI: 10.1016/j.biotechadv.2013.08.005. View

2.
Michielse C, Hooykaas P, Van Den Hondel C, Ram A . Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet. 2005; 48(1):1-17. DOI: 10.1007/s00294-005-0578-0. View

3.
Cao Q, Lin Z . Antitumor and anti-angiogenic activity of Ganoderma lucidum polysaccharides peptide. Acta Pharmacol Sin. 2004; 25(6):833-8. View

4.
Kimura T, Takeda S, Kyozuka J, Asahi T, Shimamoto K, Nakamura K . The presequence of a precursor to the delta-subunit of sweet potato mitochondrial F1ATPase is not sufficient for the transport of beta-glucuronidase (GUS) into mitochondria of tobacco, rice and yeast cells. Plant Cell Physiol. 1993; 34(2):345-55. View

5.
Wang J, Guo L, Zhang K, Wu Q, Lin J . Highly efficient Agrobacterium-mediated transformation of Volvariella volvacea. Bioresour Technol. 2008; 99(17):8524-7. DOI: 10.1016/j.biortech.2008.03.007. View