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Mycology 2017
September 25-26, 2017
conferenceseries
.com
September 25-26, 2017 Chicago, USA
2
nd
International Conference on
Mycology & Mushrooms
Volume 3, Issue 2 (Suppl)
Med Mycol Open Access
ISSN: 2471-8521
Construction of mutant strains with inducible mating type genes to identify the A mating pathway in
Flammulina velutipes
Irum Mukhtar, Aron Osakina, Baogui Xie
and
Arend F Van Peer
Fujian Agriculture and Forestry University, China
M
ating is the initial step of sexual development in higher fungi and involves the fusion of two compatible mycelia, which brings
together complementary sets of mating type genes; HD and PR mating type genes. This pairing of complementary HD and
PR genes results in activation of the respective A (HD) and B (PR) mating type pathways. Although HD and PR mating type genes
have been identified in many fungi, their downstream pathways have remained largely unknown. The present study aims to develop a
tetracycline inducible gene based system for basidiomycetous fungi, followed by the construction of mutants with inducible HD and
PR mating type genes to study these pathways in detail. We successfully adapted the doxycycline (a tetracycline derivative) inducible
promoter system (TET-On) using two expression units in a single vector; the reverse tetracycline transactivator (rtTA2S-M2) gene
that encodes the doxycycline binding TetR protein, and the tetO7 (TetR binding) sequence fused with a minial
gpd
promoter to control
expression of the gene of interest. The functionality of this system was confirmed in
F. velutipes
with two different reporter genes, one
for a Green Fluorescent Protein (gfp) and one for a Red (dTomato). Next, we selected complementary HD and PR mating type genes
for
F. velutipes
strain L11, that were subsequently cloned in the doxycycline inducible system. The first doxycycline inducible HD
mating type mutants have recently been obtained and were confirmed by qPCR. These mutants are currently being analyzed, while
transformation experiments for inducible PR and HD+PR
F. velutipes
mutants are underway.
Biography
Irum Mukhtar has expertise in fungal taxonomy, genetics and plant pathology. Her passion in fungal genetics and skill is improving information about mating in
higher fungi. Her current experimental work is based on inducible system to in study detail mating genes and downstream pathways involve in A and B pathways in
Flammulina velotipes
in which initial results showed that this approach is responsive in determining pathways in
F. velotipes
and can be used in other higher fungi.
erumm21@gmail.comIrum Mukhtar et al., Med Mycol Open Access, 3:2, 2017
DOI: 10.21767/2471-8521-C1-002