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A simple and efficient genetic transformation method of Ganoderma weberianum
YP. Zhou, MH. Chen, JJ. Lu, X. Kang, QH. Chen, XL. Huang, CE. Tian,
Language English Country United States
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Agrobacterium tumefaciens genetics metabolism MeSH
- Ganoderma genetics MeSH
- Genetic Techniques * MeSH
- Genetic Vectors genetics metabolism MeSH
- Transformation, Genetic * MeSH
- Green Fluorescent Proteins MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
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 provided by Crossref.org
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