Abstract
With genome sequences of more and more fungi become available, high-throughput systematic mutagenesis is desirable for functional genomics studies. While a number of random insertional mutagenesis and targeted gene disruption approaches have been used in filamentous fungi, Agrobacterium tumefaciens-mediated Transformation (ATMT) remains one of the most effective methods for identifying genes required for specific fungal developmental or infection processes. Because of its simplicity, ATMT is suitable for large-scale insertion mutagenesis in fungi. Magnaporthe oryzae, the rice blast fungus is a model for studying host–pathogen interactions. Here, we describe protocols for generating a M. oryzae mutant library consisting of over 70,000 ATMT transformants and for identifying genes disrupted by T-DNA in the mutants by TAIL-PCR.
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Acknowledgments
This work was supported by the 973 program (Grant No. 2006CB101901) and by the Innovative Project of State Key Laboratory for Agrobiotechnology (Grant No. 2008SKLAB0103) to Y. -L. Peng from the Ministry of Sciences and Technology, China.
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Chen, XL., Yang, J., Peng, YL. (2011). Large-Scale Insertional Mutagenesis in Magnaporthe oryzae by Agrobacterium tumefaciens-Mediated Transformation. In: Xu, JR., Bluhm, B. (eds) Fungal Genomics. Methods in Molecular Biology, vol 722. Humana Press. https://doi.org/10.1007/978-1-61779-040-9_16
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DOI: https://doi.org/10.1007/978-1-61779-040-9_16
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