- An efficient Agrobacterium-mediated transformation method for aflatoxin generation fungus Aspergillus flavus
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Guomin Han , Qian Shao , Cuiping Li , Kai Zhao , Li Jiang , Jun Fan , Haiyang Jiang , Fang Tao
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J. Microbiol. 2018;56(5):356-364. Published online May 2, 2018
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DOI: https://doi.org/10.1007/s12275-018-7349-3
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Aspergillus flavus often invade many important corps and
produce harmful aflatoxins both in preharvest and during
storage stages. The regulation mechanism of aflatoxin biosynthesis
in this fungus has not been well explored mainly
due to the lack of an efficient transformation method for
constructing a genome-wide gene mutant library. This challenge
was resolved in this study, where a reliable and efficient
Agrobacterium tumefaciens-mediated transformation (ATMT)
protocol for A. flavus NRRL 3357 was established. The results
showed that removal of multinucleate conidia, to collect
a homogenous sample of uninucleate conidia for use as the
transformation material, is the key step in this procedure.
A. tumefaciens strain AGL-1 harboring the ble gene for zeocin
resistance under the control of the gpdA promoter from
A. nidulans is suitable for genetic transformation of this fungus.
We successfully generated A. flavus transformants with
an efficiency of ~ 60 positive transformants per 106 conidia
using our protocol. A small-scale insertional mutant library
(~ 1,000 mutants) was constructed using this method and
the resulting several mutants lacked both production of conidia
and aflatoxin biosynthesis capacity. Southern blotting
analysis demonstrated that the majority of the transformants
contained a single T-DNA insert on the genome. To the best
of our knowledge, this is the first report of genetic transformation
of A. flavus via ATMT and our protocol provides an
effective tool for construction of genome-wide gene mutant
libraries for functional analysis of important genes in A.
flavus.
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Citations
Citations to this article as recorded by  -
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- Isolation and Evaluation of Terrestrial Fungi with Algicidal Ability from Zijin Mountain, Nanjing, China
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Guomin Han , Xiaoguang Feng , Yong Jia , Congyan Wang , Xingbing He , Qiyou Zhou , Xingjun Tian
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J. Microbiol. 2011;49(4):562-567. Published online September 2, 2011
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DOI: https://doi.org/10.1007/s12275-011-0496-4
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Approximately 60 fungal isolates from Zijin Mountain (Nanjing, China) were screened to determine their
algicidal ability. The results show that 8 fungi belonging to Ascomycota and 5 belonging to Basidiomycota
have algicidal ability. Of these fungi, Irpex lacteus T2b, Trametes hirsuta T24, Trametes versicolor F21a,
and Bjerkandera adusta T1 showed strong algicidal ability. The order of fungal chlorophyll-a removal efficiency
was as follows: T. versicolor F21a > I. lacteus T2b > B. adusta T1 > T. hirsuta T24. In particular, T. versicolor
F21a completely removed algal cells within 30 h, showing the strongest algicidal ability. The results also
show that all 4 fungal species degraded algal cells through direct attack. In addition, most of the tested
fungi from the order Polyporales of Basidiomycota exhibited strong algicidal activity, suggesting that most
fungi that belong to this order have algicidal ability. The findings of this work could direct the search
for terrestrial fungi for bloom control.
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