Agrobacterium tumefaciens-mediated transformation of filamentous fungi
Agrobacterium tumefaciens-mediated transformation of filamentous fungi
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DOI：
10.1038/nbt0998-839
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发表时间：
1998-09-01
影响因子：
46.9
通讯作者：
Beijersbergen, AGM
中科院分区：
文献类型：
--
作者：
de Groot, MJA;Bundock, P;Beijersbergen, AGM
根癌农杆菌在肿瘤发生过程中将其 Ti 质粒的一部分（T-DNA）转移到植物细胞中，通常用于多种植物物种的遗传修饰。我们报道根癌农杆菌还可以将其 T-DNA 有效地转移到丝状真菌泡盛曲霉中，证明了原核生物和丝状真菌之间的 DNA 转移。与泡盛曲霉原生质体转化的传统技术相比，我们转化原生质体和分生孢子的频率提高了 600 倍，大多数泡盛曲霉转化体含有随机整合在染色体位点上的单个 T-DNA 拷贝，T-DNA 以与植物描述类似的方式整合到泡盛曲霉基因组中，我们还转化了多种其他泡盛曲霉。 丝状真菌，包括黑曲霉、Fusarium venenatum、里氏木霉、胶孢炭疽菌、粗糙脉孢菌和蘑菇双孢蘑菇，表明使用根癌农杆菌的转化通常适用于丝状真菌。
Agrobacterium tumefaciens transfers part of its Ti plasmid, the T-DNA, to plant cells during tumorigenesis, It is routinely used for the genetic modification of a wide range of plant species. We report that A. tumefaciens can also transfer its T-DNA efficiently to the filamentous fungus Aspergillus awamori, demonstrating DNA transfer between a prokaryote and a filamentous fungus. We transformed both protoplasts and conidia with frequencies that were improved up to 600-fold as compared with conventional techniques for transformation of A. awamori protoplasts, The majority of the A. awamori transformants contained a single T-DNA copy randomly integrated at a chromosomal locus, The T-DNA integrated into the A. awamori genome in a manner similar to that described for plants, We also transformed a variety of other filamentous fungi, including Aspergillus niger, Fusarium venenatum, Trichoderma reesei, Colletotrichum gloeosporioides, Neurospora crassa, and the mushroom Agaricus bisporus, demonstrating that transformation using A. tumefaciens is generally applicable to filamentous fungi.