Fungi are an economic very important class of microbes. Not only do they host a range of versatile enzymes used in industrial applications (biofuels, laundry, food processing), as well do they produce several very important pharmaceutical drugs (statins and penicillins). Moreover, fungal pathogens can cause great damage in agricultural production (Phytophthora and Botrytis) and during mammalian infections (Penicillium marneffei and Candida). Transformation of DNA is used to understand the genetic basis behind these traits. Several different techniques have been developed over the years and readily shown to be decisive methods to improve fungal biotechnology. This book will cover the basics behind the most commonly used transformation methods, as well as associated tools and techniques. Each chapter will provide protocols along with examples to be used in laboratories worldwide.

Anastomosis and Heterokaryon Formation.- Induction of the Sexual Cycle in Filamentous Ascomycetes.- What Have We Learned by Doing Transformations in Neurospora tetrasperma?.- Repeat-induced Point Mutation: A Fungal-specific, Endogenous Mutagenesis Process.- Calculating RIP Mutation in Fungal Genomes Using RIPCAL.- Fungal Transposable Elements.- In vivo Targeted Mutagenesis in Yeast using TaGTEAM.- RNA Silencing in Filamentous Fungi: from Basic to Applications.- RNAi-mediated Gene Silencing in the Beta-lactam Producers Fungi Penicillium chrysogenum and Acremonium chrysogenum.- Controlling Fungal Gene Expression Using the Doxycycline-Dependent Tet-ON System in Aspergillus fumigates.- Expanding the Repertoire of Selectable Markers for Aspergillus Transformation.- Arginase (agaA) as a Fungal Transformation Marker.- Transformation of Ascomycetous Fungi using Autonomously Replicating Vectors.- A Recyclable and Bidirectionally Selectable Marker System for Transformation of Trichoderma.- Split Marker-Mediated Transformation and Targeted Gene Disruption in Filamentous Fungi.- Integrated Automation for Continuous High-Throughput Synthetic Chromosome Assembly and Transformation to Identify Improved Yeast Strains for Industrial Production of Biofuels and Bio-based Chemicals.- Imaging Flow Cytometry and High-throughput Microscopy for Automated Macroscopic Morphological Analysis of Filamentous Fungi.- Yeast Cell Electroporation in Droplet-Based Microfluidic Chip.- Identification of T-DNA Integration Sites: TAIL-PCR and Sequence Analysis.- Genetic and Genomic Manipulation in Aspergillus niger.- Genetic Manipulation of Meyerozyma guilliermondii.

Marco A. van den Berg, PhD

Principal Scientist Applied Biochemistry and Screening

DSM Biotechnology Center

Delft, The Netherlands

 Karunakaran Maruthachalam, PhD

Current: Research Scientist

Global Marker Technology Lab, DuPont-Pioneer

DuPont Knowledge Center

Andhra Pradesh, India

 Previous: Molecular Biologist

California Seed and Plant Labs

Pleasant Grove, CA, USA