Varma / Kharkwal Symbiotic Fungi

Principles and Practice
2009
ISBN: 978-3-540-95894-9
Verlag: Springer Berlin Heidelberg
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 18, 430 Seiten

Reihe: Soil Biology

Symbiotic Fungi - Principles and Practice presents current protocols for the study of symbiotic fungi and their interactions with plant roots, such as techniques for analyzing nutrient transfer, ecological restoration, microbial communication, and mycorrhizal bioassays, AM inoculum procedures and mushroom technology. The protocols offer practical solutions for researchers and students involved in the study of symbiotic microorganisms. The volume will be of great use for basic research, biotechnological applications, and the development of commercial products.

Autoren/Hrsg.

Varma, Ajit

Kharkwal, Amit C.

Inhaltsverzeichnis

1;Foreword;6
1.1;References;7
2;Preface;9
3;Contents;11
4;Contributors;14
5;Symbiosis: The Art of Living;21
5.1;1.1 Introduction;21
5.2;1.2 History of Symbiosis;22
5.3;1.3 Symbiosis of Bacteria 1.3.1 Symbiotic Association of Bacteria with Leguminous Plants;24
5.4;1.3.2 Symbiotic Association of Bacteria with Nonleguminous Plants;26
5.5;1.3.3 Establishment of the Mutualistic Relationship Between Rhizobia and Legumes;26
5.6;1.4 Symbiosis of Actinomycetes;32
5.7;1.5 Symbiosis Between Blue–Green Alga and Fungus: Geosiphon pyriforme;33
5.8;1.6 Symbiosis Between Algae and Fungi: Lichens;35
5.9;1.7 Symbiosis in Bryophytes;35
5.10;1.8 Symbiosis in Pteridophytes;36
5.11;1.9 Symbiosis in Gymnosperms;36
5.12;1.9.1 Coralloid Roots of Cycas;37
5.13;1.10 Symbiosis in Angiosperms;37
5.14;1.10.1 Mycorrhizal Symbiosis;37
5.15;1.11 Conclusions;45
5.16;References;45
6;Analysis of Rhizosphere Fungal Communities Using rRNA and rDNA;49
6.1;2.1 Introduction;49
6.2;2.2 Materials 2.2.1 Equipment;50
6.3;2.2.2 Materials;51
6.4;2.2.3 Procedure;51
6.5;2.3 Results 2.3.1 Nucleic Acid Extraction and Reverse Transcription from the Andropogon gerardii Rhizosphere;54
6.6;2.3.2 Community Assessment Using Reverse-Transcribed cDNAs and Environmental rDNAs;55
6.7;2.4 Conclusions;58
6.8;References;59
7;Use of Mycorrhiza Bioassays in Ecological Studies;61
7.1;3.1 Introduction;61
7.2;3.2 Assessment of Infectivity of AM Fungi in Soil;62
7.3;3.2.1 Direct Assessment of AM Fungi;62
7.4;3.2.2 Indirect Assessment: Mycorrhiza Bioassay;64
7.5;3.3 Purpose of Bioassays for Infectivity Assessment of AM Fungi;66
7.6;3.4 Conclusions;67
7.7;References;67
8;In Vivo Model Systems for Visualisation, Quantification and Experimental Studies of Intact Arbuscular Mycorrhizal Networks;71
8.1;4.1 Introduction;71
8.2;4.2 Structure of Pre-symbiotic Mycelium of AM Fungi: Visualisation and Quantification of Anastomosis 4.2.1 Occurrence and Frequency of Anastomosis;72
8.3;4.2.2 Dynamics of Anastomosis Formation in Living Hyphae;73
8.4;4.2.3 Cytochemical Analyses of Anastomosing Hyphae;75
8.5;4.2.4 Remarks;76
8.6;4.3 Visualisation and Quantification of Intact Mycelial Networks Spreading from Mycorrhizal Roots 4.3.1 Experimental System;76
8.7;4.3.2 Quantification of the Extent and Structure of the Mycorrhizal Network;78
8.8;4.3.3 Viability of the Mycorrhizal Network;79
8.9;4.3.4 Remarks;79
8.10;4.4 Visualisation of Belowground Links Between Plants of Different Species, Genera and Families 4.4.1 Experimental System;80
8.11;4.4.2 Occurrence and Frequency of Anastomoses Within and Between Mycorrhizal Networks;80
8.12;4.4.3 Remarks;82
8.13;4.5 Conclusions;82
8.14;References;83
9;Measurement of Net Ion Fluxes Using Ion- Selective Microelectrodes at the Surface of Ectomycorrhizal Roots;85
9.1;5.1 Introduction;85
9.2;5.2 Principle of Ion Activities Measurement with Ion- Selective Microelectrodes 5.2.1 What Is an Ion- Selective Microelectrode?;86
9.3;5.2.2 Expression of the Voltage Difference Across the Cocktail;87
9.4;5.3 Principle of Flux Measurement 5.3.1 Expression of Local Diffusion Flux in a Solution;88
9.5;5.3.2 Estimation of Ion Fluxes at the Root Surface;89
9.6;5.4 Equipment and Microelectrode Fabrication 5.4.1 Experimental Set- up;90
9.7;5.4.2 Making the Microelectrodes;93
9.8;5.4.3 Internal Silanization of the Pulled Borosilicate Tube or Microelectrode;93
9.9;5.4.4 Backfilling the Pipette;94
9.10;5.4.5 Calibration;95
9.11;5.4.6 Microelectrode Selectivity;96
9.12;5.5 Setting up the Electrophysiological Measurements;96
9.13;5.5.1 Determination of the Distances from the Root Surface for the Measurement Points;97
9.14;5.5.2 Factors Affecting the Calculation of the Concentration;99
9.15;5.5.3 Using a Complex Perfusing Solution;102
9.16;5.6 A Case Study: Measurement of NO3;103
9.17;Net Fluxes;103
9.18;into Ectomycorrhizal Short Roots 5.6.1 Validation of Flux Measurements into Coniferous Plants;103
9.19;5.6.2 Variation of NO3;105
9.20;Net Fluxes into Ectomycorrhizal Short;105
9.21;Roots: Effect of N Source Supplied to the Plants;105
9.22;5.7 Conclusions;106
9.23;References;107
10;Assessment of Phosphatase Activity Associated with Mycorrhizal Fungi by Epi- Fluorescent Microscopy;109
10.1;6.1 Introduction;109
10.2;6.2 Phosphatase Activity;110
10.3;6.3 Equipment and Laboratory Material 6.3.1 Equipment;110
10.4;6.3.2 Laboratory Material;110
10.5;6.3.3 Solutions;111
10.6;6.3.4 Protocol for Assessment of Phosphatase Activity with ELF- 97 Substrate;112
10.7;6.4 Various Applications of ELF-97 Substrate in Mycorrhizal Research;115
10.8;6.4.1 Phosphatase Activity Associated with Ectomycorrhizal Fungi;115
10.9;6.4.2 Phosphatase Activity Associated with Arbuscular Mycorrhizal Fungi;115
10.10;6.4.3 Other Applications of the ELF-97 Substrate;116
10.11;6.5 Advantages and Limitation of ELF-97 Substrate;117
10.12;6.6 Conclusions;118
10.13;References;118
11;In Vitro Compartmented Systems to Study Transport in Arbuscular Mycorrhizal Symbiosis;120
11.1;7.1 Introduction;120
11.2;7.2 Equipment and Laboratory Material 7.2.1 Equipment;121
11.3;7.2.2 Laboratory Material;121
11.4;7.3 Culture Media 7.3.1 Composition;122
11.5;7.3.2 Stock Solutions;123
11.6;7.3.3 Preparation of Culture Media;123
11.7;7.4 Transport Studies with Root-Organ Cultures (ROC);124
11.8;7.4.1 Adding Medium to the Petri Plates;125
11.9;7.4.2 Selection of a Root-Organ;126
11.10;7.4.3 Inoculation of the Root with AM Fungal Propagules;127
11.11;7.5 Transport Studies with Autotrophic Plants;128
11.12;7.5.1 Surface-Sterilization/Scarification;129
11.13;7.5.2 Medium;130
11.14;7.5.3 The Half-Closed Arbuscular MycorrhizalÒPlant ( HAMÒ P) In Vitro Culture System ( Voets et al. 2005);130
11.15;7.5.4 The Arbuscular MycorrhizalÒ Plant (AMÒP) In Vitro Culture System ( Dupre • de Boulois et al. 2006);132
11.16;7.6 Labelling with Isotopic Tracers;135
11.17;7.6.1 Stable or Radio-Isotopic Tracer?;136
11.18;7.6.2 Adding the Isotopic Tracer;137
11.19;7.6.3 Exposure Time;137
11.20;7.6.4 Harvest;139
11.21;7.7 Conclusions;139
11.22;References;139
12;Use of the Autofluorescence Properties of AM Fungi for AM Assessment and Handling;142
12.1;8.1 Introduction;142
12.2;8.2 Equipment and Laboratory Material 8.2.1 Equipment;143
12.3;8.2.2 Laboratory Material;143
12.4;8.3 Sample Preparation 8.3.1 Whole Root Samples;144
12.5;8.3.2 Root Section Samples;144
12.6;8.3.3 Isolation of Intraradical Fungal Structures;145
12.7;8.3.4 Spores;146
12.8;8.4 Bright-Field Microscopy, Epifluorescence Microscopy and Lambda- Scan;146
12.9;8.5 Autofluorescence of AM Fungal Structures;149
12.10;8.6 Viability of Fungal Structures;153
12.11;8.7 Autofluorescence Localization;155
12.12;8.8 Spore Autofluorescence and Flow Cytometry;156
12.13;8.9 Conclusions;158
12.14;References;158
13;Role of Root Exudates and Rhizosphere Microflora in the Arbuscular Mycorrhizal Fungi- Mediated Biocontrol of Phytophthora nicotianae in Tomato;160
13.1;9.1 Introduction;160
13.2;9.2 Zoospore Chemotaxy in P. nicotianae;161
13.3;9.3 Biocontrol Mediated by AMF on P. nicotianae Infecting Tomato;162
13.4;9.4 Induction of Tomato Plant Defense Mechanisms Following Mycorrhizal Colonization;162
13.5;9.5 Effect of Mycorrhizal Root Exudates on P. nicotianae Zoospore Chemotaxy In Vitro;163
13.6;9.6 Effect of Mycorrhizal Root Exudates on Tomato Infection by P. nicotianae in Soil;165
13.7;9.7 Effect of Mycorrhizal Root Exudates on the Rhizosphere Bacterial Community Structure;166
13.8;9.7.1 Antagonistic Potential of Bacteria Associated with Spores of G. mosseae;168
13.9;9.7.2 Other Mechanisms Contributing to the Biocontrol Induced by AMF on P. nicotianae;169
13.10;9.8 Conclusions;170
13.11;References;170
14;Assessing the Mycorrhizal Diversity of Soils and Identification of Fungus Fruiting Bodies and Axenic Cultures;178
14.1;10.1 Introduction;178
14.2;10.2 General Characteristics of Mycorrhizae;179
14.3;10.3 Classical Fungal Processing and Identification 10.3.1 Field Notes, Processing, Fungal Identification;179
14.4;10.3.2 Herbarium Facilities;181
14.5;10.4 Isolation of Fungal Cultures from Soils, Mycorrhizosphere and Sporophores 10.4.1 Preparations;183
14.6;10.4.2 Isolation from Fruiting Bodies and Spores;183
14.7;10.4.3 Indirect Methods for Screening;184
14.8;10.4.4 Soil Dilution Technique for Enumeration of Fungi;184
14.9;10.4.5 Direct Plate Technique;186
14.10;10.4.6 WetÒSieving and Decanting Technique for the Extraction of Spores of AMF ( Fig. 10.4);186
14.11;10.4.7 Techniques for Large Volumes of Soil;187
14.12;10.5 Preservation and Maintenance 10.5.1 Culture Collections;188
14.13;10.5.2 Cultures in Herbaria;189
14.14;10.6 Modern Molecular Methods Used in Fungal Identification;189
14.15;10.6.1 ÎÎExtraction-FreeÌÌ Preparation of PCR-Ready Material;192
14.16;10.6.2 PEX Extraction;192
14.17;10.6.3 Choice of PCR Target;197
14.18;10.6.4 PCR Troubleshoot;198
14.19;10.6.5 Sequencing and Editing Sequences;199
14.20;10.6.6 Cloning;200
14.21;10.6.7 Database Queries and Alignment;201
14.22;10.6.8 Phylogenetic Placement;202
14.23;10.7 Conclusions;203
14.24;References;203
15;Isolation of Metabolically Active Arbuscules and Intraradical Hyphae from Mycorrhizal Roots;208
15.1;11.1 Introduction;208
15.2;11.2 Isolation 11.2.1 Isolation of Arbuscules and Intraradical Hyphae with Enzymatic Digestion;209
15.3;11.2.2 Isolation of Arbuscules and Intraradical Hyphae without Enzymatic Digestion;211
15.4;11.2.3 Measurement of Metabolic Activity of Isolated Arbuscules and Hyphae;211
15.5;11.3 Conclusions;213
15.6;References;213
16;Interaction with Soil Microorganisms;215
16.1;12.1 Introduction;215
16.2;12.2 Fungal Growth Promotion in Bacterium–Fungus Co- Cultures: An Indication of Mycorrhiza Helper Function;216
16.3;12.3 Rapid Fungal Responses to Bacteria and Their Metabolites: Bacterium– Fungus Suspension Cultures;218
16.4;12.4 Materials 12.4.1 Reagents for Culture;219
16.5;12.4.2 Organisms;220
16.6;12.5 Procedures 12.5.1 Culture on Solid Media;221
16.7;12.5.2 Suspension Cultures;222
16.8;12.5.3 Co-Cultures;223
16.9;12.5.4 Simple Culture System for the Inoculation of Norway Spruce Roots with Actinomycetes;224
16.10;12.5.5 Impact of Pre-Inoculation with Mycorrhization Helper Bacteria on Heterobasidion Root Rot;224
16.11;12.5.6 Impact of Volatiles of Mycorrhization Helper Bacteria on Heterobasidion Root Rot;225
16.12;12.5.7 Physiological Screening of Host Plant Viability;225
16.13;References;226
17;Isolation, Cultivation and In Planta Visualization of Bacterial Endophytes in Hanging Roots of Banyan Tree ( Ficus bengalensis);229
17.1;13.1 Introduction;229
17.2;13.2 Isolation and Cultivation of Endophytes from Plant Roots;230
17.3;13.2.1 Selection of Plant Material;230
17.4;13.2.2 Isolation of Endophytic Microorganisms;230
17.5;13.2.3 Cultivationof Endophytes;232
17.6;13.3 Protocol for Isolation and Cultivation of Bacterial Endophytes from Hanging Roots of Banyan Tree 13.3.1 Requirements;232
17.7;13.3.2 Media for Cultivation;233
17.8;13.3.3 Method;233
17.9;13.4 In Planta Visualization/Localization of Endophytes;234
17.10;13.4.1 Vital Staining Method;234
17.11;13.4.2 Electron Microscopy;235
17.12;13.5 Protocol for Detection of Endophytic Bacteria by Vital Staining;235
17.13;13.5.1 Equipment;235
17.14;13.5.2 Chemicals and Reagents;236
17.15;13.5.3 Procedure;236
17.16;13.6 Protocol for Detection of Bacterial Endophytes by Transmission Electron Microscopy;237
17.17;13.6.1 Requirements;237
17.18;13.6.2 Procedure;239
17.19;13.7 Conclusions;241
17.20;References;242
18;Micro-PIXE Analysis for Localization and Quantification of Elements in Roots of Mycorrhizal Metal- Tolerant Plants;244
18.1;14.1 Introduction;244
18.2;14.2 Materials and Procedures 14.2.1 Equipment for Sample Preparation;245
18.3;14.2.2 Laboratory Materials;245
18.4;14.2.3 Specimen Preparation;245
18.5;14.2.4 Micro-PIXE Analysis;249
18.6;14.3 Example of Micro-PIXE Analysis;252
18.7;14.3.1 Sample Preparation and Micro-PIXE Analysis;252
18.8;14.3.2 Results;257
18.9;14.4 Conclusions;258
18.10;References;258
19;Functional Genomic of Arbuscular Mycorrhizal Symbiosis: Why and How Using Proteomics;260
19.1;15.1 Introduction;261
19.2;15.2 General Considerations About the Biological Material;262
19.3;15.2.1 Root Handling for Protein Extraction Protocols of;262
19.4;15.3 Materials and Apparatus 15.3.1 Products and Buffers for Protein Extraction;263
19.5;15.3.2 Products and Buffers for 2DE;264
19.6;15.3.3 Products for Protein Staining;265
19.7;15.3.4 Equipment;265
19.8;15.4 Procedure;268
19.9;15.4.1 Extraction of Total and Soluble Proteins;268
19.10;15.4.2 Combined Extraction of RNA and Proteins;269
19.11;15.4.3 Protein Extraction;270
19.12;15.4.4 Measure of Protein Content in Samples;270
19.13;15.4.5 Two-Dimensional Electrophoresis;271
19.14;15.4.6 Protein Detection;275
19.15;15.5 A Case Study Applied to Dissect the Early Stages of Medicago Truncatula Mycorrhizal Symbiosis: When Transcriptomics and Proteomics are Working Together;281
19.16;15.5.1 In Silico Analysis;282
19.17;15.5.2 Proteomic Analysis;283
19.18;15.6 Conclusions;286
19.19;15.7 New Insights;287
19.20;References;288
20;Using Stable Carbon Isotope Labelling in Signature Fatty Acids to Track Carbon Allocation in Arbuscular Mycorrhiza;292
20.1;16.1 Introduction;292
20.2;16.2 Microbial Biomass Distribution;293
20.3;16.3;294
20.4;C Labelling 16.3.1 Timing of Measurements;294
20.5;16.3.2 Labelling in Monoxenic AM Cultures;294
20.6;16.3.3 Labelling in Pot Experiments;295
20.7;16.3.4 Labelling in the Field;295
20.8;16.4 Analytical Techniques 16.4.1 Homogenization of Samples;296
20.9;16.4.2 Lipid Analysis;296
20.10;16.4.3 Signature Fatty Acids;296
20.11;16.4.4 Determination of;297
20.12;C Enrichment in Crude Tissue;297
20.13;Samples and Fatty Acids;297
20.14;16.4.5 Quantification of Transferred Carbon;298
20.15;16.5 Sensitivity and Specificity of the Method;298
20.16;16.6 Conclusions;299
20.17;References;300
21;N Enrichment Methods to Quantify Two-Way Nitrogen Transfer Between Plants Linked by Mychorrhizal Networks;302
21.1;17.1 Introduction;302
21.2;17.2 Use of;303
21.3;N Isotopes for Investigating Nitrogen;303
21.4;Transfer Between Plants;303
21.5;17.3 Experimental Design for Investigating Two-Way Nitrogen Transfer Between Plants;304
21.6;17.3.1 Quantification of Nitrogen Transfer Between Plants;305
21.7;17.4 Conclusions;307
21.8;References;307
22;Analyses of Ecophysiological Traits of Tropical Rain Forest Seedlings Under Arbuscular Mycorrhization: Implications in Ecological Restoration;309
22.1;18.1 Introduction;309
22.2;18.1.1 Fragmentation;309
22.3;18.1.2 Restoration;310
22.4;18.1.3 What is the Role of Arbuscular Mycorrhizal Fungi in Habitat Recovery?;310
22.5;18.2 Objective;311
22.6;18.3 Case Study 18.3.1 Study Site;312
22.7;18.3.2 Methods;313
22.8;18.3.3 Results;314
22.9;18.3.4 Remarks;315
22.10;18.4 Restoration and Arbuscular Mycorrhizae Fungi;318
22.11;18.5 Conclusions;319
22.12;References;319
23;Techniques for Arbuscular Mycorrhiza Inoculum Reduction;322
23.1;19.1 Introduction;322
23.2;19.2 Solarization;323
23.3;19.3 Steam Sterilization;324
23.4;19.4 Pasteurization;325
23.5;19.5 Gamma (g)-Irradiation;326
23.6;19.6 Chemicals;326
23.7;19.7 Soil Disturbance;328
23.8;19.8 Crop Rotation;329
23.9;19.9 Other Methods;329
23.10;19.10 Conclusions;330
23.11;References;331
24;Best Production Practice of Arbuscular Mycorrhizal Inoculum;334
24.1;20.1 Introduction;334
24.2;20.2 Working Hypotheses (Following Feldmann 1998);335
24.3;20.3 Basic Assumptions for the Inoculum Production ( Following Feldmann and Grotkass 2002);335
24.4;20.3.1 The Planning Phase: Define what you Need;336
24.5;20.3.2 The Analytical Phase: Make your AMF Isolate a Strain and Describe its Abilities;338
24.6;20.3.3 The Adaption Phase: Direction Instead of Screenings;341
24.7;20.3.4 The Up-Scaling Phase: One Further Step Only;342
24.8;20.4 Conclusions;347
24.9;References;350
25;The Use of AMF and PGPR Inoculants Singly and Combined to Promote Microplant Establishment, Growth and Health;352
25.1;21.1 Introduction;352
25.2;21.2 Mode of Action and Safety of Inoculants;354
25.3;21.2.1 Arbuscular Mycorrhizal Fungi;354
25.4;21.2.2 Plant Growth Promoting Rhizobacteria (PGPR);355
25.5;21.3 Selection, Production and Formulation of Inoculants;357
25.6;21.3.1 Arbuscular Mycorrhizal Fungi;357
25.7;21.3.2 Plant Growth-Promoting Rhizobacteria;358
25.8;21.4 Use of Inoculants in Micropropagation 21.4.1 Arbuscular Mycorrhizal Fungi;360
25.9;21.4.2 Plant Growth-Promoting Rhizobacteria;364
25.10;21.5 Holistic Strategies for the Use of Inoculants;365
25.11;21.6 Conclusions;367
25.12;References;367
26;Co-Culture of Linum album Cells and Piriformospora indica for Improved Production of Phytopharmaceuticals;376
26.1;22.1 Introduction;376
26.2;22.2 Development of Plant Cell Cultures 22.2.1 Germination of Seeds;378
26.3;22.2.2 Initiation of Callus Cultures;378
26.4;22.2.3 Initiation of Suspension Cultures;379
26.5;22.3 Development of Fungal Culture 22.3.1 Maintenance of Piriformospora indica;379
26.6;22.3.2 Initiation of Fungal Culture;380
26.7;22.4 Establishment of Co-Culture of L. album with P. indica 22.4.1 Development of Co- Culture ( Fig. 22.1);382
26.8;22.5 Analysis 22.5.1 Growth in Terms of Dry Cell Weight;383
26.9;22.5.2 Determination of Fungal and Plant Biomass;383
26.10;22.5.3 Extraction and Estimation of Podophyllotoxin;384
26.11;22.5.4 Phenylalanine Ammonia Lyase (PAL) Enzyme Extraction and Assay;385
26.12;22.6 Conclusions;385
26.13;References;385
27;Fungal Elicitors for Enhanced Production of Secondary Metabolites in Plant Cell Suspension Cultures;388
27.1;23.1 Introduction;388
27.2;23.2 Development of Plant Cell Cultures 23.2.1 Germination of Seeds;390
27.3;23.2.2 Initiation of Callus Cultures;390
27.4;23.2.3 Initiation of Suspension Cultures;391
27.5;23.3 Development of Elicitors 23.3.1 Preparation of Elicitors;391
27.6;23.3.2 Addition of Elicitors;392
27.7;23.4 Analysis 23.4.1 Growth in Terms of Dry Cell Weight;393
27.8;23.4.2 Extraction and Estimation of Podophyllotoxin;394
27.9;23.5 Conclusions;394
27.10;References;394
28;Auxin Production by Symbiotic Fungi: Bioassay and HPLC- MS Analysis;396
28.1;24.1 Introduction;396
28.2;24.2 Equipment and Laboratory Material 24.2.1 Equipment;397
28.3;24.2.2 Laboratory Material;398
28.4;24.3 Interaction on the Same Medium: Piriformospora/ Arabidopsis 24.3.1 Preparation of Slope Agar Plates;398
28.5;24.3.2 Inoculation of the Fungus;400
28.6;24.3.3 Adding Arabidopsis Seeds to the Petri Dish;400
28.7;24.3.4 Growth Conditions;402
28.8;24.3.5 Evaluation of Roots;402
28.9;24.4 Interaction on Two Different Media: Truffles/ Arabidopsis;402
28.10;24.4.1 Preparation of Dual Bioassay Plates;402
28.11;24.4.2 Inoculation of the Fungus;402
28.12;24.4.3 Adding Arabidopsis Seeds to the Petri Dish;403
28.13;24.4.4 Growth Conditions and Evaluation;403
28.14;24.4.5 Estimation of the IAA Amount Produced by the Fungus;404
28.15;24.4.6 IAA Quantification in Agar Plates;404
28.16;24.4.7 Extraction of the Agar Plates for IAA Determination;405
28.17;24.4.8 HPLC-ESI-MS/MS Determination of IAA;405
28.18;24.5 Examples of IAA Production by Fungi 24.5.1 Piriformospora indica;406
28.19;24.5.2 Truffles;407
28.20;References;407
29;Siderophores of Mycorrhizal Fungi: Detection, Isolation and Identification;408
29.1;25.1 Introduction;408
29.2;25.2 Isolation of Hydroxamate Siderophores from Fungal Culture Filtrates;409
29.3;25.3 Chemical Assays;412
29.4;25.3.1 Chrome Azurol S (CAS) Assay ( Modified After Schwyn and Neilands 1987);412
29.5;25.4 Separation of Ferric Hydroxamates by HPLC;413
29.6;25.5 Mass Spectrometry;414
29.7;25.5.1 Gas Chromatography: Mass Spectrometry;414
29.8;25.6 NMR Spectroscopy;414
29.9;25.7 Conclusions;415
29.10;References;416
30;Biology and Molecular Approaches in Genetic Improvement of Cultivated Button Mushroom ( Agaricus Bisporus);418
30.1;26.1 General Biology 26.1.1 Morphology and Life Cycle;418
30.2;26.1.2 Taxonomy and Nomenclature;420
30.3;26.1.3 Nutritional Requirements;423
30.4;26.1.4 Environmental Requirements;424
30.5;26.1.5 Sexuality and Breeding;426
30.6;26.2 Trouble Shooting;433
30.7;References;434
31;Index;437