By John F. Leslie and Brett A. Summerell

John F. Leslie received his Ph.D. in genetics from the University of Wisconsin, Madison in 1979 and is currently a Professor of Plant Pathology and Genetics at Kansas State University.

Brett A. Summerell received his Ph.D. in Plant Pathology from the University of Sydney in 1988 at which time he moved to the Royal Botanic Gardens (Sydney, Australia), where he currently is the Director of Science and Public Programs.

For the first time in over 20 years, a comprehensive collection of photographs and descriptions of species in the fungal genus Fusarium is available. The Fusarium Laboratory Manual provides an overview of the biology of Fusarium and the techniques involved in the isolation, identification, and characterization of individual species and the populations in which they occur. It is the first time that genetic, morphological, and molecular approaches have been incorporated into a volume devoted to Fusarium identification.

This book is designed as a laboratory manual and a guide to techniques and species likely to be encountered. This manual includes some of the evolutionary biology and population genetics thinking that has begun to inform the understanding of agriculturally important fungal pathogens. In addition, it provides guidance in formulating questions and obtaining answers about this very important group of fungi. The need for as many different techniques as possible to be used in the identification and characterization process has never been greater. These approaches have applications to fungi other than those in the genus Fusarium. This volume presents an introduction to the genus Fusarium, the toxins these fungi produce, and the diseases they can cause.

Key features:

• Descriptions of 70 species, both new and old
• Practical "how-to" protocols for genetic, morphological, and molecular identification techniques
• Recipes for many commonly used media
• Techniques that will work for common field and molecular applications
• Complete index and references

Species descriptions include:

• Diagram of sexual stages
• Common synonyms
• Reported geographic and host distribution
• Media to examine and key characters: Characters on CLA and PDA; Macroconidia, Microconidia, and Chlamydospores
• Taxonomy, pathology, and ecology

The Fusarium Laboratory Manual

1. Introduction

Techniques and Methods

2. Media - Recipes and Preparation

2.1 Media for Growing and Identifying Fusarium
2.2 Supplementary Identification Media
2.3 Media for Isolating Fusarium
2.4 Media for the Preparation of Natural Inocula
2.5 Synthetic and Semi-synthetic Media
2.6 Media for Sexual Crosses
2.7 Sterilization of Media and Materials

3. Techniques for Recovering Fusarium

3.1 Collecting strategy(ies)
3.2 Isolation Techniques - Plants
3.3 Isolation Techniques - Soil
3.4 Isolation Techniques - Spore Trapping and Air Sampling
3.5 Seed Disinfestation

4. Techniques for Growing and Maintaining Fusarium

4.1 Vegetative Propagation
4.2 Preparing Cultures for Identification
4.3 Single Spore Subcultures
4.4 Mutagenesis
4.5 Culture Preservation

5. Vegetative Compatibility Groups (VCGs)

5.1 History of and Genetic Basis Underlying Vegetative Compatibility
5.2 Overall Strategy for Determining if Strains are Vegetatively Compatible
5.3 Recovering and Identifying nit Mutants
5.4 Typical Pairing Protocols
5.5 Common Trouble Spots - HSI, crn, and NitMs
5.6 Characterizing a Population with VCGs

6. Fertility Concepts

6.1 Heterothallic, Homothallic and Pseudohomothallic
6.2 Mating Type
6.3 Population Effects of Mating Type
6.4 Male, Female, and Hermaphrodite
6.5 Crossing Protocols
6.6 Developing Female-Fertile Tester Strains
6.7 Species Identification Through Sexual Crosses

7. Nucleic Acid Analyses

7.1 DNA Extraction and Purification
7.2 PCR - Mating-Type Alleles
7.3 Amplified Fragment Length Polymorphisms (AFLPs)
7.4 Sequence Analysis and Sequenced Loci
7.5 Genetic Maps

Taxonomy and Identification of Fusarium

8. A Brief History of Fusarium Taxonomy

9. Species Concepts in Fusarium

9.1 Generic Problems in Speciation in Fusarium
9.2 Morphological Species Concepts
9.3 Biological Species Concepts
9.4 Phylogenetic Species Concepts
9.5 How Many Strains Make a Species?
9.6 Species Names
9.7 Subspecific Terminology
9.8 A Species Concept for Fusarium

10. Teleomorphs of Fusarium

10.1 Taxonomy of Teleomorphs
10.2 General Teleomorph Characters
10.3 Sexual Development and Differentiation
10.4 Spore Killer
10.5 Anamorph-Teleomorph Connections

11. Practical Approaches to Identification

11.1 Overall Identification Strategy
11.2 The Diseased Plant and Its Geographic Origin
11.3 Native and Agricultural Populations
11.4 Culture Preparation
11.5 The Essence of Morphological Identifications
11.6 Beyond Morphology - Sexual Cross Fertility
11.7 Beyond Morphology - Molecular Diagnostics
11.8 The Special Case of Fusarium oxysporum
11.9 Differences Between Temperate and Tropical Regions
11.10 Conclusions

Species Descriptions

12. Morphological Characters

12.1 Macroconidia
12.2 Microconidia
12.3 Chlamydospores
12.4 Other Characters
12.5 Secondary Characters

13. Species Descriptions

F. acuminatum
F. acutatum
F. andiyazi
F. anthophilum
F. armeniacum
F. avenaceum
F. aywerte
F. babinda
F. begoniae
F. beomiforme
F. brevicatenulatum
F. bulbicola
F. camptoceras
F. chlamydosporum
F. circinatum
F. compactum
F. concentricum
F. crookwellense (F. cerealis)
F. culmorum
F. decemcellulare
F. denticulatum
F. dimerum
F. dlamini
F. equiseti
F. foetens
F. fujikuroi
F. globosum
F. graminearum
F. guttiforme
F. heterosporum
F. hostae
F. konzum
F. lactis
F. lateritium
F. longipes
F. mangiferae
F. merismoides
F. miscanthi
F. musarum
F. napiforme
F. nelsonii
F. nisikadoi
F. nurragi
F. nygamai
F. oxysporum
F. phyllophilum
F. poae
F. polyphialidicum
F. proliferatum
F. pseudoanthophilum
F. pseudocircinatum
F. pseudograminearum
F. pseudonygamai
F. ramigenum
F. redolens
F. sacchari
F. sambucinum
F. scirpi
F. semitectum (F. incarnatum)
F. solani
F. sporotrichioides
F. sterilihyphosum
F. subglutinans
F. succisae
F. thapsinum
F. torulosum
F. tricinctum
F. udum
F. venenatum
F. verticillioides

“It was lovely to read the detailed techniques and methods included in this book. Taking into account all informative aspects and practical instructions which have been integrated in this text, The Fusarium Laboratory Manual would indeed assist as a guidebook for hands-on workshops as well as a guide for those who are looking for a reference, description of a common species, or a technique especially suited with such an important genus Fusarium.”
—Fungal Diversity

"The detail given for techniques is extensive, almost anticipating any trouble one might encounter."
—Inoculum

"The Fusarium Laboratory Manual is a milestone in the study of the genus Fusarium and will help bridge the gap between morphological and phylogenetic taxonomy. It will be used by everybody dealing with Fusarium in the Third Millennium."
—W.F.O. Marasas, Medical Research Council, South Africa