An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae)
Abstract 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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Sociedade Brasileira de Genética
2021
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oai:scielo:S1415-475720210004004022021-09-28An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae)Sbaraini,NicolauTomazett,Mariana VieiraPenteriche,Augusto BartzGonçales,Relber AguiarCamargo,Matheus da SilvaBailão,Alexandre MeloBorges,Clayton LuizSchrank,AugustoSoares,Célia Maria de AlmeidaStaats,Charley Christian Simplicillium Simplicillium subtropicum Agrobacterium tumefaciens Katushka Agrobacterium tumefaciens-mediated transformation Abstract 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.info:eu-repo/semantics/openAccessSociedade Brasileira de GenéticaGenetics and Molecular Biology v.44 n.3 20212021-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572021000400402en10.1590/1678-4685-gmb-2021-0073 |
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Sbaraini,Nicolau Tomazett,Mariana Vieira Penteriche,Augusto Bartz Gonçales,Relber Aguiar Camargo,Matheus da Silva Bailão,Alexandre Melo Borges,Clayton Luiz Schrank,Augusto Soares,Célia Maria de Almeida Staats,Charley Christian |
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Sbaraini,Nicolau Tomazett,Mariana Vieira Penteriche,Augusto Bartz Gonçales,Relber Aguiar Camargo,Matheus da Silva Bailão,Alexandre Melo Borges,Clayton Luiz Schrank,Augusto Soares,Célia Maria de Almeida Staats,Charley Christian An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
author_facet |
Sbaraini,Nicolau Tomazett,Mariana Vieira Penteriche,Augusto Bartz Gonçales,Relber Aguiar Camargo,Matheus da Silva Bailão,Alexandre Melo Borges,Clayton Luiz Schrank,Augusto Soares,Célia Maria de Almeida Staats,Charley Christian |
author_sort |
Sbaraini,Nicolau |
title |
An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
title_short |
An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
title_full |
An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
title_fullStr |
An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
title_full_unstemmed |
An efficient Agrobacterium tumefaciens-mediated transformation method for Simplicillium subtropicum (Hypocreales: Cordycipitaceae) |
title_sort |
efficient agrobacterium tumefaciens-mediated transformation method for simplicillium subtropicum (hypocreales: cordycipitaceae) |
description |
Abstract 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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Sociedade Brasileira de Genética |
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2021 |
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http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572021000400402 |
work_keys_str_mv |
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