An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae)
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2021
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Filamentous fungi are the organisms of choice for most industrial biotechnology. Some species can produce a variety
of secondary metabolites and enzymes of commercial interest, and the production of valuable molecules has been
enhanced through different molecular tools. Methods for genetic manipulation and transformation have been essential
for the optimization of these organisms. The genus Simplicillium has attracted increased attention given several
potential biotechnological applications. The Simplicillium genus harbors several entomopathogenic species and some
isolates have been explored for bioremediation of heavy metal contaminants. Furthermore, the myriad of secondary
metabolites isolated from Simplicillium spp. render these organisms as ideal targets for deep exploration and further
biotechnological mining possibilities. However, the lack of molecular tools hampered the exploration of this genus.
Thus, an Agrobacterium tumefaciens-mediated transformation method was established for Simplicillium subtropicum,
employing the far-red fluorescent protein TURBOFP635/Katushka, as a visual marker, and the selection marker SUR
gene, that confers resistance to chlorimuron ethyl. Notably, one round of transformation using the established method
yielded almost 400 chlorimuron resistant isolates. Furthermore, these transformants displayed mitotic stability for, at
least, five generations. We anticipate that this method can be useful for deep molecular exploration and improvement
of strains in the Simplicillium genus.
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Simplicillium, Simplicillium subtropicum, Agrobacterium tumefaciens, Katushka, Agrobacterium tumefaciens-mediated transformation
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SBARAINI, Nicolau et al. An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae). Genetics and Molecular Biology, Ribeirão Preto, v. 44, n. 3, e20210073, 2021. DOI: 10.1590/1678-4685-GMB-2021-0073. Disponível em: https://www.scielo.br/j/gmb/a/6qhqqN3qCD3Ljdpw5VXZm4n/. Acesso em: 8 ago. 2024.