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Efficient Transformation of the Medicinal
Mushroom Ganoderma lucidum
LI SUN, HUAQING CAI, WEIHONG XU, YUANLEI HU, YIN GAO and
National Key Laboratory of Protein Engineering and Plant Genetic Engineering,
College of Life Sciences, Peking University, Beijing 100871, China
Abstract. Ganoderma lucidum is a well-known and important medicinal mushroom, but
its genetic modification has not been reported. We developed an efficient procedure for
isolation and regeneration of protoplasts from G. lucidum. To construct a vector for high-
level expression of heterologous genes in G. lucidum, the 1.4-kb regulatory region of the
glyceraldehyde-3-phosphate dehydrogenase gene (GPD) was isolated from the genomic
DNA of Lentinus edodes, and the GPD promoter was fused to the
and bialaphos resistance (bar) genes. Using the resulting construct, p301-bG1, an efficient
transformation system based on electroporation was established for G. lucidum. GUS ex-
pression was observed among transformants conferring bialaphos resistance, indicating
that the L. edodes GPD promoter can be used for expression of exogenous genes in G.
lucidum. We also studied green fluorescent protein (GFP) as another reporter for transfor-
mation of G. lucidum. The L. edodes GPD promoter was fused respectively to the GFP
and bar genes, and the resulting construct, p301-bg, was introduced into G. lucidum. Sta-
ble GFP expression in transformants was detectable by fluorescence microscopy, suggest-
ing the suitability of GFP as a reporter system in transformation of this mushroom. This is
the first report of an efficient transformation system for G. lucidum using different report-
ers, paving the way for genetic modification of this famous medicinal mushroom.
: This manuscript, in detail, is available only in the electronic version of the
Key words: bialaphos resistance, Ganoderma lucidum, GFP, GPD promoter, GUS,
Abbreviations: bar, bialaphos resistance; BSA, bovine serum albumen; GFP, green fluo-
rescent protein; GPD, glyceraldehyde-3-phosphate dehydrogenase; LMP, low melting
point; nos, nopaline synthase.
Transformation of Ganoderma lucidum Sun et al.
Plant Molecular Biology Reporter 19: 383a–383j, December 2001
© 2001 International Society for Plant Molecular Biology. Printed in Canada.
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ph: (86-10) 6275-3062.
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