Main description:

The kingdom Fungi constitutes an independent group equal in rank to that of plants and animals. It is a diverse clade of heterotrophic eukaryotic organisms that shares some ch- acteristics with animals and includes mushrooms, molds, yeasts as well as many other types of less well known organisms. Approximately 100,000 species have been described, which comprise less than 10% of the estimated number of fungal species in nature. Fungi can be found in every place wherever adequate moisture, temperature, and organic substrates are available; however, they also occupy extreme habitats, from hot volcanoes to arctic zones, arid deserts, and deep oceans. The importance of fungi as a group is tremendous; most species are saprobes and play prime roles in decomposition and the recycling of organic matter and nutrients, and many of them produce enzymes and metabolites with important applications in pharmacology, biotechnology, and other industries. Alongside the positive aspects, fungi also cause huge damage, primarily as plant pathogens. Fungi are highly amenable to molecular work, and a few fungal species serve as model systems to study basic processes with results that are applicable to many organisms, including humans.

Contents:

Part I: Fungal Transformation and Gene Knockout

1. Protoplast Transformation of Filamentous Fungi
Gillian B. Turgeon, Bradford Condon, Jinyuan Liu, and Ning Zhang

2. Electroporation and Agrobacterium-Mediated Spore Transformation
Anna Minz and Amir Sharon

3. High-Throughput Construction of Gene Deletion Cassettes for Generation of Neurospora crassa Knockout Strains
Patrick D. Collopy, Hildur V. Colot, Gyungsoon Park, Carol Ringelberg, Christopher M. Crew, Katherine A. Borkovich, and Jay C. Dunlap

4. Development of Impala-Based Transposon Systems for Gene Tagging in Filamentous Fungi
Marie Dufresne and Marie-Josee Daboussi

5. DelsGate: A Robust and Rapid Method for Gene Deletion
Maria D. Garcia-Pedrajas, Marina Nadal, Timothy Denny, Lourdes Baeza-Montanez, Zahi Paz, and Scott E. Gold

6. Gene Silencing for Functional Analysis: Assessing RNAi as a Tool for Manipulation of Gene Expression
Carmit Ziv and Oded Yarden

Part II: Detection and Quantification of Fungi

7. Analysis of Fungal Gene Expression by Real Time Quantitative PCR
Shahar Ish-Shalom and Amnon Lichter

8. Identification of Differentially Expressed Fungal Genes in planta by Suppression Subtraction Hybridization (SSH)
Benjamin A. Horwitz and Sophie Lev

9. Quantification of Fungal Infection of Leaves with Digital Images and Scion Image Software
Paul H. Goodwin and Tom Hsiang

10. Expression Profiling of Fungal Genes During Arbuscular Mycorrhiza Symbiosis Establishment Using Direct Fluorescent In situ RT-PCR
Pascale M. A. Seddas-Dozolme, Christine Arnould, Marie Tollot, Elena Kuznetsova, and Vivienne Gianinazzi-Pearson

11. Application of Laser Microdissection to Study Plant-Fungal PathogenInteractions
John Fosu-Nyarko, Michael G.K. Jones, and Zhaohui Wang

12. Multiplex Gene Expression Analysis by TRAC in Fungal Cultures
Jari J. Rautio

13. Amplification of Fungal Genomes Using Multiple Displacement Amplification
Simon J. Foster and Brendon J. Monahan

Part III: Microscopy and Protein Analysis

14. Biochemical Methods Used to Study the Gene Expression and Protein Complexes in the Filamentous Fungus Neurospora crassa
Jinhu Guo, Guocun Huang, Joonseok Cha, and Yi Liu

15. Measuring Protein Kinase and Sugar Kinase Activity in Plant Pathogenic Fusarium Species
Burton H. Bluhm and Xinhua Zhao

16. A Detailed Protocol for Chromatin Immunoprecipitation (ChIP) in the Yeast Saccharomyces cerevisiae
Melanie Grably and David Engelberg

17. A Method to Visualize the Actin and Microtubule Cytoskeleton by Indirect Immunofluorescence
Flora Banuett

18. Fluorescence in situ Hybridization for Molecular Cytogenetic Analysis in Filamentous Fungi
Dai Tsuchiya and Masatoki Taga

19. Live-Cell Imaging of Microtubule Dynamics in Hyphae of Neurospora crassa
Maho Uchida, Rosa R. Mourino-Perez, and Robert W. Roberson

20. Methods to Detect Apoptotic-Like Cell Death in Filamentous Fungi
Camile P. Semighini and Steven D. Harris

21. Evaluation of Antifungal Susceptibility Using Flow Cytometry
Cidalia Pina-Vaz and Acacio Goncalves Rodrigues

22. Preparation of Fungi for Ultrastuctural Investigations and Immunogoldlabelling
Gerd Hause and Simone Jahn

23. Split-EGFP Screens for the Detection and Localization of Protein-Protein Interactions in Living Yeast Cells
Emma Barnard and David J. Timson