A silver bullet in a golden age of functional genomics: the impact of Agrobacterium-mediated transformation of fungi.
A silver bullet in a golden age of functional genomics: the impact of Agrobacterium-mediated transformation of fungi.
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DOI：
10.1186/s40694-017-0035-0
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发表时间：
2017
影响因子：
--
通讯作者：
Jeon J
中科院分区：
文献类型：
--
作者：
Idnurm A;Bailey AM;Cairns TC;Elliott CE;Foster GD;Ianiri G;Jeon J
根癌农杆菌作为转化工具的实施彻底改变了在大量真菌物种中发现和理解基因功能的方法。A.根癌农杆菌介导的转化（AtMT）是近20年来发展起来的真菌研究中最具变革性的技术之一，可以说，基因组学的影响超越了这一发展。AtMT已被广泛应用于正向遗传学，其中使用随机T-DNA插入诱变结合表型筛选产生菌株文库，使得许多过程的遗传基础得以阐明。或者，AtMT是反向遗传学的基础，在反向遗传学中，产生具有靶基因缺失或破坏的突变体分离株，从而能够确定基因的功能作用。当与基因组学的同步进展相结合时，使用AtMT的正向和反向方法都使复杂的真菌表型能够在分子和遗传水平上被解剖。此外，在一些情况下，AtMT已经为新物种的开发铺平了道路，这些新物种可以作为真菌生物学特定领域的模型，特别是在植物病原性子囊菌和一些担子菌物种中。尽管它的影响，AtMT的实施一直是不平衡的真菌。这篇评论提供了深入了解新的研究工具扩展到一个大型的研究社区和跨多个生物体的动态。因此，真菌中的AtMT，除了已经证明的和持续的基因发现能力之外，作为一种简单的转化工具，还提供了一个模型来了解其他刚刚开创的技术，例如CRISPR/Cas，如何在真菌和其他真核生物物种中发挥作用。
The implementation of Agrobacterium tumefaciens as a transformation tool revolutionized approaches to discover and understand gene functions in a large number of fungal species. A. tumefaciens mediated transformation (AtMT) is one of the most transformative technologies for research on fungi developed in the last 20 years, a development arguably only surpassed by the impact of genomics. AtMT has been widely applied in forward genetics, whereby generation of strain libraries using random T-DNA insertional mutagenesis, combined with phenotypic screening, has enabled the genetic basis of many processes to be elucidated. Alternatively, AtMT has been fundamental for reverse genetics, where mutant isolates are generated with targeted gene deletions or disruptions, enabling gene functional roles to be determined. When combined with concomitant advances in genomics, both forward and reverse approaches using AtMT have enabled complex fungal phenotypes to be dissected at the molecular and genetic level. Additionally, in several cases AtMT has paved the way for the development of new species to act as models for specific areas of fungal biology, particularly in plant pathogenic ascomycetes and in a number of basidiomycete species. Despite its impact, the implementation of AtMT has been uneven in the fungi. This review provides insight into the dynamics of expansion of new research tools into a large research community and across multiple organisms. As such, AtMT in the fungi, beyond the demonstrated and continuing power for gene discovery and as a facile transformation tool, provides a model to understand how other technologies that are just being pioneered, e.g. CRISPR/Cas, may play roles in fungi and other eukaryotic species.