de Groot M J, Bundock P, Hooykaas P J, Beijersbergen A G
Unilever Research Laboratory Vlaardingen, The Netherlands.
Nat Biotechnol. 1998 Sep;16(9):839-42. doi: 10.1038/nbt0998-839.
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.
根癌土壤杆菌在致瘤过程中将其Ti质粒的一部分，即T-DNA，转移到植物细胞中。它通常用于多种植物物种的基因改造。我们报告称，根癌土壤杆菌还能将其T-DNA高效转移至丝状真菌泡盛曲霉，这证明了原核生物与丝状真菌之间的DNA转移。与传统的泡盛曲霉原生质体转化技术相比，我们用该方法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单个随机整合到染色体位点的T-DNA拷贝。T-DNA以与植物中描述的方式相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丝状真菌，包括黑曲霉、产毒镰刀菌、里氏木霉、炭疽菌、粗糙脉孢菌和双孢蘑菇，这表明利用根癌土壤杆菌进行转化普遍适用于丝状真菌。
