Advanced techniques in soil microbiology:
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| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Berlin ; Heidelberg ; New York
Springer
2007
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| Series: | Soil biology
11 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Literaturangaben |
| Physical Description: | XXVIII, 427 S. Ill., graph. Darst. 24 cm |
| ISBN: | 9783540708643 3540708642 |
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| 245 | 1 | 0 | |a Advanced techniques in soil microbiology |c Ajit Varma ... (ed.) |
| 264 | 1 | |a Berlin ; Heidelberg ; New York |b Springer |c 2007 | |
| 300 | |a XXVIII, 427 S. |b Ill., graph. Darst. |c 24 cm | ||
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| 490 | 1 | |a Soil biology |v 11 | |
| 500 | |a Literaturangaben | ||
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| _version_ | 1804137255861747712 |
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| adam_text | Contents
1 Detection and Diversity of Fungi from Environmental Samples:
Traditional Versus Molecular Approaches ......................... 1
R. Jeewon and K.D. Hyde
1.1 Introduction ............................................... 1
1.2 Microscopy and Culture-Based Methods ....................... 2
1.3 Molecular-Based Methods .................................... 4
1.4 The Nuclear-Encoded Ribosomal DNA Gene:
Phylogenetic and Systematic Value .......................... 5
1.5 Denaturing Gradient Gel Electrophoresis:
Applicability, Usefulness and Bias ......................... 7
1.6 Conclusions and Future Directions ......................... 11
References ...................................................... 11
2 Functional Genomic Approaches for Mycorrhizal Research ........... 17
A. K. Pandey, H. White, and G.K. Podila
2.1 Introduction .............................................. 17
2.2 Yeast Two Hybrid: An Approach
for Understanding Signaling Pathways ...................... 18
2.3 Agrobacterium-Medialed Transformation in Laccaria bicolor ... 22
2.4 Materials and Methods ..................................... 24
2.4.1 Interaction Studies of Laccaria bicolor with Aspen
(Populus tremuloides) Seedlings .................... 24
2.4.2 Yeast Two-Hybrid Protocol .......................... 26
2.4.3 Agrobacterium-Mediated Transformation in Laccaria
bicolor ............................................ 28
References ...................................................... 30
3 Automated Fluoroscence Sequencing and Troubleshooting............. 35
S. Gochhait, D. Malhotra, E. Rai, and R.N.K. Bamezai
3.1 Introduction .............................................. 35
3.2 Evolution of the Method ................................... 36
3.2.1 Manual Sequencing .................................. 36
XII_____________________________________________________________Contents
3.2.2 Automated Sequencing .............................. 37
3.2.3 Pyrosequencing..................................... 39
3.3 Methods .................................................. 39
3.3.1 Template Preparation .............................. 39
3.3.2. Reaction Setup (BigDye Terminator Cycle Sequencing) 40
3.3.3 Performing Cycle Sequencing ....................... 41
3.3.4 Preparing Extension Products for Electrophoresis .. 42
3.4 Trouble Shooting ......................................... 43
3.4.1 Problem: Flat Line or “Dead On Analysis” .......... 43
3.4.2 Problem: Noisy Data (Background) .................. 45
3.4.3 Problem: Reading Near the Primer .................. 46
3.4.4 Problem: Strong Terminator Peaks .................. 46
3.4.5 Problem: Low Intensity of Shorter Products ........ 48
3.4.6 Problem: Longer Fragments Missing ................. 49
3.4.7 Problem: Presence of Spikes ....................... 49
3.4.8 Problem: Weaker Signals ........................... 49
References ..................................................... 50
4 mRNA Quantitation Using Real Time PCR .......................... 53
S. Gochhait, S.I. Bukhari, and R.N.K. Bamezai
4.1 Introduction ............................................. 53
4.2 Methods .................................................. 54
4.2.1 Chemistry and Primer/Probe Design ................. 54
4.2.2 RNA Isolation from the Sample ..................... 57
4.2.3 Reverse Transcription ............................. 58
4.2.4 Real Time PCR Set Up .............................. 59
4.2.5 Instrumentation ................................... 61
4.2.6 Data Analysis ..................................... 67
4.3 Notes .................................................... 68
References ..................................................... 71
5 Laboratory Practice for the Production of Polyclonal
and Monoclonal Antibodies ...................................... 73
S. Khurana, S. Bhaskar, and A. Mukhopadhyay
5.1 Introduction ............................................. 73
5.2 Production of Polyclonal Antibodies ...................... 74
5.2.1 Choice of Animal and Method of Immunization ....... 74
5.2.2 Preparation for Immunization ...................... 75
5.2.3 Production of Polyclonal Antibodies ............... 76
5.2.4 Materials and Equipment ........................... 77
5.3 Production of Monoclonal Antibodies ...................... 77
5.3.1 Immunization of Mice or Rats ...................... 77
5.3.2 Myeloma Cell Culture .............................. 78
5.3.3 Setup for Fusion of Myeloma with Spleen Cells ..... 79
5.3.4 Selection and Cloning of Hybridoma ................ 79
Contents
XIII
5.3.5 Production of Monoclonal Antibodies ................ 80
5.3.6 Materials and Equipment ............................ 81
5.4 Purification of Antibody .................................. 82
5.4.1 Purification of IgG by Precipitation with Ammonium
Sulfate ............................................ 83
5.4.2 Purification of IgG by DEAE-Sepharose
Chromatography...................................... 83
5.4.3 Purification of IgG Using Immobilized Protein A .... 84
5.5 Analysis of Purity of IgG by Electrophoresis .............. 86
5.5.1 Materials and Equipment ............................ 88
5.6 Enzyme-Linked Immunosorbent Assay ..................... 88
5.6.1 Materials and Equipment ............................ 90
5.7 Conclusion ................................................ 90
References ...................................................... 90
6 Modern Techniques for Analyzing Immunological Responses .......... 93
Satish Khurana, Sangeeta Bhaskar, and Asok Mukhopdhyay
6.1 Introduction .............................................. 93
6.2 Type of Immune Responses .................................. 93
6.2.1 Innate Immune Response ............................. 94
6.2.2 Adaptive Immune Response ........................... 94
6.3 Adaptive Immune System .................................... 94
6.3.1 Humoral Immune System .............................. 95
6.3.2 Cellular Immune System ............................. 96
6.4 Different Assay Systems to Study the Adaptive Immune
Response .................................................. 96
6.4.1 Mixed Lymphocyte Proliferation Assays .............. 96
6.4.2 Detection of Type of T Helper Responses (Thl/Th2) .. 98
6.4.3 Cytotoxic T Lymphocyte Activity .................... 99
6.5 Flow Cytometric Analysis of Immune Cells.................. 102
6.5.1 Materials and Equipment ........................... 105
6.6 Magnetic Activated Cell Sorting .......................... 105
6.6.1 Materials and Equipment ........................... 107
6.7 Isolation of Mononuclear Cells from Peripheral Blood ..... 107
6.7.1 Materials and Equipment ........................... 108
6.8 Conclusions .............................................. 108
References ..................................................... 108
7 TranscriptomeAnalysis ........................................... Ill
S. K. Yadav, S.L. Singla-Pareek, and A. Pareek
7.1 Introduction ............................................. Ill
7.2 RNA Preparation .......................................... 113
7.3 Northern Analysis ........................................ 113
7.3.1 Principle ......................................... 113
7.3.2 Procedure ......................................... 114
XIV
Contents
7.3.3 Applications ....................................... 116
7.4 In Situ Hybridization ...................................... 116
7.4.1 Principle .......................................... 116
7.4.2 Procedure .......................................... 117
7.4.3 Applications ....................................... 119
7.5 Dot Blot and Slot Blot ..................................... 120
7.5.1 Principle .......................................... 120
7.5.2 Procedure .......................................... 120
7.5.3 Applications........................................ 123
7.6 Reverse Transcriptase-Polymerase Chain Reaction ............ 123
7.6.1 Principle .......................................... 123
7.6.2 Procedure .......................................... 124
7.6.3 Application ofRT-PCR ............................... 125
7.7 DNA Microarray ............................................. 126
7.7.1 Principle .......................................... 126
7.7.2 Procedure .......................................... 127
7.7.3 Applications ....................................... 129
7.8 Conclusions ................................................ 129
References ....................................................... 130
8 RNAi Technology: a Tool for Functional Validation of Novel Genes 133
R. Karan, S. Kumari, S.K. Yadav, and A. Pareek
8.1 Introduction ............................................. 133
8.2 Machinery Involved in RNAi ............................... 134
8.2.1 Inducer ........................................... 135
8.2.2 Dicer ............................................. 135
8.2.3 RNA-Dependent RNA Polymerase ...................... 135
8.2.4 RNA-Induced Silencing Complex ..................... 135
8.2.5 miRNA and siRNA ................................... 136
8.3 RNAi as a Tool of Functional Genomics .................... 136
8.3.1 Production of dsRNA................................ 137
8.3.2 Constitutive and Inducible RNAi ................... 138
8.3.3 Antisense RNA and RNAi ............................ 140
8.4 Potential Areas of Application ........................... 140
8.5 Conclusions .............................................. 142
References .................................................... 142
9 Molecular Matchmaking: Techniques for Biomolecular
Interactions ................................................... 145
R. Oberoi, P. Kumar, and S.K. Lai
9.1 Introduction .............................................. 145
9.2 Tools for the Study of Protein-Protein Interactions ....... 145
9.2.1 The Two-Hybrid System .............................. 147
9.2.2 The Split-Ubiquitin System ......................... 148
Contents
XV
9.2.3 Reverse Two-Hybrid System ......................... 148
9.2.4 Sos Recruitment System
(Cyto Trap Yeast Two-Hybrid System) ............... 148
9.2.5 Yeast One-Hybrid System .......................... 149
9.2.6 Double Interaction Screen ......................... 149
9.2.7 Yeast Three-Hybrid or Tri-Hybrid System ........... 150
9.3 Procedure ................................................ 150
9.3.1 Reagents, Materials, and Equipment ................ 151
9.3.2 Notes and Points to Watch ......................... 152
References ..................................................... 152
10 Environmental Proteomics: Extraction and Identification
of Protein in Soil ............................... 155
Z. Solaiman, M. A. Kashem and I. Matsumoto
10.1 Introduction ............................................. 155
10.2 Sample Preparations ...................................... 156
10.3 Protocols for Protein Extraction from Soil ............... 157
10.3.1 Extraction of Extracellular Protein ............... 157
10.3.2 Extraction of Whole-Cell Protein .................. 157
10.4 Protein Loading .......................................... 158
10.5 Protein Expression Analyses .............................. 158
10.5.1 SDS-PAGE .......................................... 158
10.5.2 Two-Dimension SDS-PAGE Analysis .................. 158
10.6. Gel Staining ............................................. 161
10.6.1 Coomassie Brilliant Blue Staining Protocol
(For Mini Gels) ................................... 162
10.6.2 Silver Staining Protocol .......................... 162
10.7 Image Analysis ........................................... 163
10.8 Spot Cut ................................................. 163
10.9 Protein Digestion ........................................ 163
10.10 Mass Spectrometry Analysis ............................... 164
10.11 Spectral Analysis ........................................ 164
10.12 N-Terminal Amino Acid Sequencing ......................... 165
10.13 Conclusions .............................................. 165
References ..................................................... 166
11 DGGE and RISA Protocols for Microbial Community Analysis
in Soil ........................................................ 167
Z. Solaiman and P. Marschner
11.1 Introduction ............................................. 167
11.2 Soil DNA Extraction ...................................... 168
11.2.1 Equipment ......................................... 168
11.2.2 Chemicals ......................................... 168
11.2.3 DNA Extraction Protocol .......................... 168
XVI
Contents
11.3 Polymerase Chain Reaction Protocol for DGGE .............. 170
11.3.1 First Round PCR ................................... 170
11.3.2 GC Clamp 16S PCR (Second Round PCR) .............. 171
11.4 DGGE Techniques .......................................... 172
11.4.1 Equipment ......................................... 172
11.4.2 Chemicals ......................................... 173
11.4.3 Assembling the Gel Chamber ....................... 173
11.4.4 Casting the Gel ................................... 174
11.4.5 Loading of the Samples ............................ 175
11.4.6 Staining and Imaging of the Gels ................. 175
11.5 Ribosomal Intergenic Spacer Analysis ..................... 175
11.5.1 Equipment ......................................... 176
11.5.2 Chemicals ......................................... 176
11.5.3 PCR Protocol ...................................... 176
11.5.4 Gel Preparation and Loading ....................... 177
11.5.5 Gel Running ....................................... 178
11.5.6 Staining and Imaging of the Gels ................. 178
11.6 Data Analysis ............................................ 178
11.7 Conclusions .............................................. 179
References .................................................... 179
12 Soil Microbial Community Structureand Function Assessed
by FAME, PLFA and DGGE ֊ Advantages and Limitations ........... 181
P. Marschner
12.1 Introduction ............................................. 181
12.2 Microbial Community Structure Based on Fatty Acid Patterns 182
12.2.1 FAME Extraction and Data Analysis ................ 183
12.2.2 PLFA Analysis ..................................... 185
12.2.3 Advantages and Limitations of Fatty Acid Patterns .... 188
12.3 Denaturing Gradient Gel Electrophoresis ........ 189
12.3.1 DNA Extraction from Soil .......................... 189
12.3.2 Polymerase Chain Reaction ......................... 190
12.3.3 DGGE Procedures ................................... 192
12.4 Conclusions .............................................. 196
References .................................................... ! 97
13 Measurement of Microbial Biomass and Activity in Soil ......... 201
Z. Solaiman
13.1 Introduction ............................................. 201
13.2 Protocols for Microbial Biomass Determination ............ 202
13.2.1 Chloroform Fumigation-Extraction Method
for Microbial Biomass C and N .................... 202
13.2.2 Hexanol Extraction Method for Microbial P ........ 205
13.3 Protocol for Total Microbial Activity Determination ...... 207
13.3.1 Equipment ......................................... 207
Contents
XVII
13.3.2 Reagents ............................................ 207
13.3.3 Protocol for Extraction ............................. 208
13.4 Protocol for Soil Dehydrogenase Enzyme Analysis ............ 208
13.4.1 Equipment ........................................... 209
13.4.2 Reagents ............................................ 209
13.4.3 Protocol for Extraction ............................. 209
13.5 Conclusions ................................................ 209
References ....................................................... 210
14 Immuno-Technology for the Localization of Acid Phosphatase
Using Native Gel Bands in Piriformospora indica and Other Soil
Microorganisms ............................................ 213
R. Malla, U. Pokharel, R. Prasad, R. Oelmüller, and A. Varma
14.1 Introduction ............................................... 213
14.1.1 Taxonomic Status .................................... 213
14.1.2 Phosphatases ........................................ 214
14.2 Immunotechnology for the Detection and Localization
of Acid Phosphatase in P. indica ........................... 216
14.2.1 Extraction of Protein and Enzyme Assay ...... 216
14.2.2 Purification of Protein by Column Chromatography .. 217
14.2.3 Purification of Protein by Ion Exchange
Chromatography........................................ 217
14.2.4 Native Polyacrylamide Gel Electrophoresis ........... 219
14.2.5 Detection of Enzyme in Native PAGE .................. 220
14.2.6 Isolation of Acid Phosphatase for Raising Antibody ... 222
14.2.7 Production of Antibodies using Acid Phosphatase
in Native Gel ........................................ 222
14.2.8 Antiserum Preparation ............................... 224
14.2.9 Purification of Immunoglobulin from Serum ........... 225
14.2.10 Western Blot ....................................... 226
14.2.11 Immuno-Fluorescence ................................ 228
14.2.12 Localization of ACPase by Immunogold Technique ... 228
14.3 Troubleshooting ............................................ 232
14.4 Conclusions ................................................ 232
References ....................................................... 232
15 Use of Short Oligonucleotide Primers in Random Amplified
Polymorphic DNA Techniques for Species Identification . 237
R. Malla and A. Varma
15.1 Introduction ............................................... 237
15.2 Polymorphism between Piriformospora indica and Sebacina
vermifera, Members of the Order Sebacinales ................ 239
15.3 General Protocol for RAPD Technique
to Show Polymorphism ....................................... 241
15.3.1 Experimental Procedures ............................. 242
XVIII
Contents
15.4 Troubleshooting .......................................... 244
15.5 Conclusions .............................................. 244
References ..................................................... 245
16 Co-Cultivation with Sebacinales ................................ 247
A.C. Kharkwal, R. Prasad, H. Kharkwal, A. Das,
K. Bhatnagar, I. Sherameti, R. Oelmiiller, and A. Varma
16.1 Introduction ............................................. 247
16.2 Sebacinaceae - Novel Fungi ............................... 248
16.3 Host Spectrum ............................................ 249
16.4 Functions of the Sebacinaceae ............................ 251
16.5 Eco-Functional Identity .................................. 252
16.6 Axenic Co-Cultivation of Sebacinaceae .................... 254
16.6.1 Procedure ......................................... 254
16.6.2 Protocol .......................................... 256
16.7 Media Compositions ....................................... 256
16.8 Seed Surface Sterilization and Germination ............... 261
16.8.1 Protocol for Seed Surface Sterilization ........... 262
16.8.2 Inoculum Placement in the Pots .................... 262
16.8.3 Results ........................................... 262
16.9 Comparative Study on Plant Growth with Treated
Endosymbionts ............................................ 264
16.10 In Vivo Co-Cultivation of Sebacinales ................... 264
16.11 Conclusions.............................................. 266
References ..................................................... 267
17 Quantitative Histochemistry: a Forgotten Tool
with New Applications .......................................... 271
R. Hampp and S. Haag
17.1 Introduction ............................................. 271
17.2 Sample Preparation and Handling .......................... 272
17.3 Microphotometry .......................................... 274
17.4 Biochemical Analysis: Real Time Microassays .............. 276
17.5 Spatial Resolution of Basic Steps
of Fungal Trehalose Metabolism in Symbiosis .............. 277
References ..................................................... 279
18 Ion Cyclotron Resonance Fourier Transform Mass Spectrometry
for Non-Targeted Metabolomics of Molecular Interactions
in the Rhizosphere ............................................. 281
P. Schmitt-Kopplin, N. Hertkorn, M. Frommberger,
M. Ludo, M. Englmann, A. Fekete, and I. Gebefugi
18.1 Introduction ............................................. 281
18.2 The Chemical Biology Approach ............................ 282
Contents
XIX
18.3 Complementary Analytical Approaches .................... 283
18.3.1 Targeted Analysis ............................... 284
18.3.2 Metabolite Profiling ............................ 285
18.3.3 Non-Targeted Analysis ........................... 285
18.4 Resolving Structural Information from Molecular Complexity
with ICR-FT/MS ......................................... 286
18.4.1 Top-Down Approach: From ICR-FT/MS-Profiling
Analysis to Structural Hypothesis ............... 288
18.4.2 Complementary Analytical Tools .................. 290
18.4.3 Bottom-Up Approach: From Hypothesis-Driven
Experiments Upwards to ICR-FT/MS ................ 290
18.5 Conclusion ............................................. 292
References .................................................. 292
19 Application of Terminal-Restriction Fragment Length
Polymorphism for MolecularAnalysis of Soil Bacterial
Communities ..................................................... 295
A. Mengoni, E. Giuntini, and M. Bazzicalupo
19.1 Introduction .............................................. 295
19.2 A General Protocol for Taxonomic T-RFLP Profiling of Soil
Bacterial Communities ..................................... 297
19.2.1 Materials .......................................... 297
19.2.2 Experimental Procedure ............................ 298
19.2.3 Troubleshooting .................................... 299
19.3 Standardization of T-RFLP Profiles ........................ 299
19.4 Other Applications of T-RFLP to Soil Bacterial Communities 301
19.5 Conclusions ............................................... 302
References ...................................................... 302
20 Molecular Symbiotic Analysis Between Arabiopsis thaliana
and Piriformospora indica ....................................... 307
B. Shahollari, K. Bhatnagar, I. Sherameti, A. Varma, and R. Oelmüller
20.1 Introduction .............................................. 307
20.2 Beneficial Interaction Between Plants and Fungi:
Piriformospora indica and Arabidopsis thaliana
as a Model System ......................................... 308
20.3 Co-Cultivation of P. indica and Arabidopsis under Standardized
Growth Conditions ......................................... 309
20.4 Map-Based Cloning of a Mutated Gene ....................... 312
20.5 Rapid DNA Extraction ...................................... 313
20.6 Confirmation of a Mutated Phenotype of an EMS Mutant
by the Analysis of an Independent T-DNA Insertion Line .... 313
20.7 Differntial Display to Identify Genes which are Regulated
in Response to P. indica .................................. 314
20.8 Activation Tagged Lines ................................... 315
XX
Contents
20.9 Identification of Biochemical Pathways
in A. thaliana which are Regulated by P. indica ........... 317
References ....................................................... 317
21 Biophysical Phenomics Reveals Functional Building Blocks
of Plants Systems Biology: a Case Study for the Evaluation
of the Impact of Mycorrhization with Piriformospora indica ....... 319
R.}, Strasser, M. Tsimilli-Michael, D. Dangre, and M. Rai
21.1 Introduction .............................................. 319
21.2 Biophysical Phenomics of the Fast Fluorescence Rise O-J-I-P 320
21.2.1 The Energy Cascade in the Photosynthetic Apparatus 320
21.2.2 Microstates - Functional Building Blocks
of Photosynthesis .................................. 320
21.2.3 Measuring Fluorescence Transients with PEA,
Handy-PEA and FIM- Fluorimeters .................... 322
21.2.4 How Fluorescence Kinetics Provide an Insight
to the Microstates - Functional Blocks of PSII ..... 323
21.3 Case Study ................................................ 332
21.3.1 Mycorrhization and the Advantages of Piriformospora
indica, an Emerging Growth Booster ................. 332
21.3.2 Phenomics of the O-J-I-P Fluorescence Transient
for the Study of Cadmium Stress on Chick Peas
(Cicer arietinum L. Chafa variety) With and Without
Symbiosis With Glomus mosseae, G. caledonium
and Piriformospora indica .......................... 333
21.3.3 Correlation of Physiological with Biophysical
Parameters ......................................... 337
21.4 Conclusions ............................................... 338
References ....................................................... 338
22 Analysis of the Plant Protective Potential of the Root Endophytic
Fungus Piriformospora indica in Cereals .......................... 343
F. Waller, B. Achatz, and K.-H. Kogel
22.1 Introduction .............................................. 343
22.2 Plant Responses and Resistance to Pathogens ............... 344
22.2.1 Local Reactions .................................... 344
22.2.2 Systemic Reactions and Resistance in Cereals ....... 344
22.2.3 Beneficial Microbial Endophytes Protecting Cereals
from Pathogens ..................................... 345
22.3 Interaction of P indica with Cereals ...................... 345
22.3.1 P. indica Colonizes Root Cortical Cells in Barley... 346
22.3.2 P. indica Enhances Biomass and Yield in Barley ..... 346
22.4 Approaches to Study the Mechanism
of P indica-Induced Pathogen Resistance.................... 347
Contents
XXI
22.4.1 P indica Induces Disease Resistance Against Root
Pathogens ........................................... 347
22.4.2 P. indica Induces Systemic Disease Resistance ....... 348
22.4.3 Assessment of the Antioxidant Capacity of P. indica-
Infested Roots ...................................... 350
22.4.4 Gene Expression Induced by P. indica in Barley Leaves 351
22.5 Conclusions ................................................ 351
References ....................................................... 352
23 Members of Sebacinales Confer Resistance Against Heavy Metal
Stress in Plants ................................................. 355
N. Hahn, A. Varma, R. Oelmüller, and I. Sherameti
23.1 Introduction ............................................... 355
23.2 Scientific Background ...................................... 355
23.3 Differential Display to Understand Cd2+ Resistance
Mediated by Endophytic Fungi ............................... 357
23.4 Studies on Protein Level ................................... 357
23.4.1 Two-Dimensional Gel Electrophoresis,
Preparation of Proteins ............................. 359
23.4.2 Mass Spectrometry, Preparation of Samples
by Tryptic Digestion ................................ 359
References ....................................................... 360
24 Screening of Plant Growth-Promoting Rhizobacteria ................ 363
C.S. Nautiyal and S.M. DasGupta
24.1 Introduction ............................................... 363
24.2 Candidature for Being a Rhizobacteria ...................... 364
24.3 Screening Methods .......................................... 365
24.3.1 Criteria for Screening .............................. 365
24.3.2 Selection of Screening Methods ...................... 365
24.3.3 Classic Methods ..................................... 366
24.3.4 Modern Methods ...................................... 368
24.3.5 Molecular Methods ................................... 370
24.4 Metagenomics ............................................... 371
24.5 Tracking of GEMs ........................................... 372
24.6 Conclusions ................................................ 372
References ....................................................... 373
25 Research Methods in Arbuscular Mycorrhizal Fungi ................. 377
A. Gaur and A. Varma
25.1 Introduction ............................................... 377
25.2 Assessment of AM Fungal Propagules in Soil ................. 378
25.2.1 Soil Sampling........................................ 378
XXII
Contents
25.2.2 Spore Extraction ................................... 378
25.2.3 Quantification of Spore Numbers ................... 379
25.2.4 Infectivity Assays ................................. 379
25.2.5 Identification of AM Fungi ........................ 380
25.2.6 Use of Fatty Acids for Identification of AM Fungi... 381
25.3 Quantification of AM Fungal Root Colonization in Root ...... 382
25.3.1 Clearing and Staining Roots ....................... 382
25.3.2 Modifications of Staining Procedure ............... 383
25.3.3 Measurement of Root Colonization by AM Fungi .... 384
25.4 Extraction and Quantification of Extra-Radical Mycelium
of AM Fungi in Soils ...................................... 384
25.5 Assessment of Growth Response of Effective Isolates ........ 385
25.6 Inoculum Production of AM Fungi ............................ 386
25.6.1 On-Farm Production of AM Fungi .................... 386
25.6.2 Traditional Culture Methods .............................. 387
25.6.3 AM Fungal Culture Using Aeroponic and Hydroponic
Culture ............................................ 388
25.6.4 Monoaxenic Culture of AM Fungi ..................... 389
25.6.5 Storage of AM Fungal Inoculum ...................... 390
25.7 Conclusions ................................................ 390
References ....................................................... 391
26 Field Trials of Bioinoculants .................................... 397
I. Orta$ and A. Varma
26.1 Introduction ............................................... 397
26.2 Effect of Mycorrhizal Infection on Nutrient Uptake ......... 398
26.3 Effect of Soil Fumigation and Mycorrhizal
Inoculation on Plant Growth Under Field Conditions ........ 399
26.4 Effect of Mycorrhizal Inoculation on Plant Growth
and Nutrient Uptake under Non-Sterile Field Conditions..... 403
26.5 Soil and Crop Management System ............................ 408
26.6 Inoculation Techniques ..................................... 409
26.7 Conclusion ................................................. 411
References ....................................................... 412
Subject Index
415
|
| adam_txt |
Contents
1 Detection and Diversity of Fungi from Environmental Samples:
Traditional Versus Molecular Approaches . 1
R. Jeewon and K.D. Hyde
1.1 Introduction . 1
1.2 Microscopy and Culture-Based Methods . 2
1.3 Molecular-Based Methods . 4
1.4 The Nuclear-Encoded Ribosomal DNA Gene:
Phylogenetic and Systematic Value . 5
1.5 Denaturing Gradient Gel Electrophoresis:
Applicability, Usefulness and Bias . 7
1.6 Conclusions and Future Directions . 11
References . 11
2 Functional Genomic Approaches for Mycorrhizal Research . 17
A. K. Pandey, H. White, and G.K. Podila
2.1 Introduction . 17
2.2 Yeast Two Hybrid: An Approach
for Understanding Signaling Pathways . 18
2.3 Agrobacterium-Medialed Transformation in Laccaria bicolor . 22
2.4 Materials and Methods . 24
2.4.1 Interaction Studies of Laccaria bicolor with Aspen
(Populus tremuloides) Seedlings . 24
2.4.2 Yeast Two-Hybrid Protocol . 26
2.4.3 Agrobacterium-Mediated Transformation in Laccaria
bicolor . 28
References . 30
3 Automated Fluoroscence Sequencing and Troubleshooting. 35
S. Gochhait, D. Malhotra, E. Rai, and R.N.K. Bamezai
3.1 Introduction . 35
3.2 Evolution of the Method . 36
3.2.1 Manual Sequencing . 36
XII_Contents
3.2.2 Automated Sequencing . 37
3.2.3 Pyrosequencing. 39
3.3 Methods . 39
3.3.1 Template Preparation . 39
3.3.2. Reaction Setup (BigDye Terminator Cycle Sequencing) 40
3.3.3 Performing Cycle Sequencing . 41
3.3.4 Preparing Extension Products for Electrophoresis . 42
3.4 Trouble Shooting . 43
3.4.1 Problem: Flat Line or “Dead On Analysis” . 43
3.4.2 Problem: Noisy Data (Background) . 45
3.4.3 Problem: Reading Near the Primer . 46
3.4.4 Problem: Strong Terminator Peaks . 46
3.4.5 Problem: Low Intensity of Shorter Products . 48
3.4.6 Problem: Longer Fragments Missing . 49
3.4.7 Problem: Presence of Spikes . 49
3.4.8 Problem: Weaker Signals . 49
References . 50
4 mRNA Quantitation Using Real Time PCR . 53
S. Gochhait, S.I. Bukhari, and R.N.K. Bamezai
4.1 Introduction . 53
4.2 Methods . 54
4.2.1 Chemistry and Primer/Probe Design . 54
4.2.2 RNA Isolation from the Sample . 57
4.2.3 Reverse Transcription . 58
4.2.4 Real Time PCR Set Up . 59
4.2.5 Instrumentation . 61
4.2.6 Data Analysis . 67
4.3 Notes . 68
References . 71
5 Laboratory Practice for the Production of Polyclonal
and Monoclonal Antibodies . 73
S. Khurana, S. Bhaskar, and A. Mukhopadhyay
5.1 Introduction . 73
5.2 Production of Polyclonal Antibodies . 74
5.2.1 Choice of Animal and Method of Immunization . 74
5.2.2 Preparation for Immunization . 75
5.2.3 Production of Polyclonal Antibodies . 76
5.2.4 Materials and Equipment . 77
5.3 Production of Monoclonal Antibodies . 77
5.3.1 Immunization of Mice or Rats . 77
5.3.2 Myeloma Cell Culture . 78
5.3.3 Setup for Fusion of Myeloma with Spleen Cells . 79
5.3.4 Selection and Cloning of Hybridoma . 79
Contents
XIII
5.3.5 Production of Monoclonal Antibodies . 80
5.3.6 Materials and Equipment . 81
5.4 Purification of Antibody . 82
5.4.1 Purification of IgG by Precipitation with Ammonium
Sulfate . 83
5.4.2 Purification of IgG by DEAE-Sepharose
Chromatography. 83
5.4.3 Purification of IgG Using Immobilized Protein A . 84
5.5 Analysis of Purity of IgG by Electrophoresis . 86
5.5.1 Materials and Equipment . 88
5.6 Enzyme-Linked Immunosorbent Assay . 88
5.6.1 Materials and Equipment . 90
5.7 Conclusion . 90
References . 90
6 Modern Techniques for Analyzing Immunological Responses . 93
Satish Khurana, Sangeeta Bhaskar, and Asok Mukhopdhyay
6.1 Introduction . 93
6.2 Type of Immune Responses . 93
6.2.1 Innate Immune Response . 94
6.2.2 Adaptive Immune Response . 94
6.3 Adaptive Immune System . 94
6.3.1 Humoral Immune System . 95
6.3.2 Cellular Immune System . 96
6.4 Different Assay Systems to Study the Adaptive Immune
Response . 96
6.4.1 Mixed Lymphocyte Proliferation Assays . 96
6.4.2 Detection of Type of T Helper Responses (Thl/Th2) . 98
6.4.3 Cytotoxic T Lymphocyte Activity . 99
6.5 Flow Cytometric Analysis of Immune Cells. 102
6.5.1 Materials and Equipment . 105
6.6 Magnetic Activated Cell Sorting . 105
6.6.1 Materials and Equipment . 107
6.7 Isolation of Mononuclear Cells from Peripheral Blood . 107
6.7.1 Materials and Equipment . 108
6.8 Conclusions . 108
References . 108
7 TranscriptomeAnalysis . Ill
S. K. Yadav, S.L. Singla-Pareek, and A. Pareek
7.1 Introduction . Ill
7.2 RNA Preparation . 113
7.3 Northern Analysis . 113
7.3.1 Principle . 113
7.3.2 Procedure . 114
XIV
Contents
7.3.3 Applications . 116
7.4 In Situ Hybridization . 116
7.4.1 Principle . 116
7.4.2 Procedure . 117
7.4.3 Applications . 119
7.5 Dot Blot and Slot Blot . 120
7.5.1 Principle . 120
7.5.2 Procedure . 120
7.5.3 Applications. 123
7.6 Reverse Transcriptase-Polymerase Chain Reaction . 123
7.6.1 Principle . 123
7.6.2 Procedure . 124
7.6.3 Application ofRT-PCR . 125
7.7 DNA Microarray . 126
7.7.1 Principle . 126
7.7.2 Procedure . 127
7.7.3 Applications . 129
7.8 Conclusions . 129
References . 130
8 RNAi Technology: a Tool for Functional Validation of Novel Genes 133
R. Karan, S. Kumari, S.K. Yadav, and A. Pareek
8.1 Introduction . 133
8.2 Machinery Involved in RNAi . 134
8.2.1 Inducer . 135
8.2.2 Dicer . 135
8.2.3 RNA-Dependent RNA Polymerase . 135
8.2.4 RNA-Induced Silencing Complex . 135
8.2.5 miRNA and siRNA . 136
8.3 RNAi as a Tool of Functional Genomics . 136
8.3.1 Production of dsRNA. 137
8.3.2 Constitutive and Inducible RNAi . 138
8.3.3 Antisense RNA and RNAi . 140
8.4 Potential Areas of Application . 140
8.5 Conclusions . 142
References . 142
9 Molecular Matchmaking: Techniques for Biomolecular
Interactions . 145
R. Oberoi, P. Kumar, and S.K. Lai
9.1 Introduction . 145
9.2 Tools for the Study of Protein-Protein Interactions . 145
9.2.1 The Two-Hybrid System . 147
9.2.2 The Split-Ubiquitin System . 148
Contents
XV
9.2.3 Reverse Two-Hybrid System . 148
9.2.4 Sos Recruitment System
(Cyto Trap Yeast Two-Hybrid System) . 148
9.2.5 Yeast One-Hybrid System . 149
9.2.6 Double Interaction Screen . 149
9.2.7 Yeast Three-Hybrid or Tri-Hybrid System . 150
9.3 Procedure . 150
9.3.1 Reagents, Materials, and Equipment . 151
9.3.2 Notes and Points to Watch . 152
References . 152
10 Environmental Proteomics: Extraction and Identification
of Protein in Soil . 155
Z. Solaiman, M. A. Kashem and I. Matsumoto
10.1 Introduction . 155
10.2 Sample Preparations . 156
10.3 Protocols for Protein Extraction from Soil . 157
10.3.1 Extraction of Extracellular Protein . 157
10.3.2 Extraction of Whole-Cell Protein . 157
10.4 Protein Loading . 158
10.5 Protein Expression Analyses . 158
10.5.1 SDS-PAGE . 158
10.5.2 Two-Dimension SDS-PAGE Analysis . 158
10.6. Gel Staining . 161
10.6.1 Coomassie Brilliant Blue Staining Protocol
(For Mini Gels) . 162
10.6.2 Silver Staining Protocol . 162
10.7 Image Analysis . 163
10.8 Spot Cut . 163
10.9 Protein Digestion . 163
10.10 Mass Spectrometry Analysis . 164
10.11 Spectral Analysis . 164
10.12 N-Terminal Amino Acid Sequencing . 165
10.13 Conclusions . 165
References . 166
11 DGGE and RISA Protocols for Microbial Community Analysis
in Soil . 167
Z. Solaiman and P. Marschner
11.1 Introduction . 167
11.2 Soil DNA Extraction . 168
11.2.1 Equipment . 168
11.2.2 Chemicals . 168
11.2.3 DNA Extraction Protocol . 168
XVI
Contents
11.3 Polymerase Chain Reaction Protocol for DGGE . 170
11.3.1 First Round PCR . 170
11.3.2 GC Clamp 16S PCR (Second Round PCR) . 171
11.4 DGGE Techniques . 172
11.4.1 Equipment . 172
11.4.2 Chemicals . 173
11.4.3 Assembling the Gel Chamber . 173
11.4.4 Casting the Gel . 174
11.4.5 Loading of the Samples . 175
11.4.6 Staining and Imaging of the Gels . 175
11.5 Ribosomal Intergenic Spacer Analysis . 175
11.5.1 Equipment . 176
11.5.2 Chemicals . 176
11.5.3 PCR Protocol . 176
11.5.4 Gel Preparation and Loading . 177
11.5.5 Gel Running . 178
11.5.6 Staining and Imaging of the Gels . 178
11.6 Data Analysis . 178
11.7 Conclusions . 179
References . 179
12 Soil Microbial Community Structureand Function Assessed
by FAME, PLFA and DGGE ֊ Advantages and Limitations . 181
P. Marschner
12.1 Introduction . 181
12.2 Microbial Community Structure Based on Fatty Acid Patterns 182
12.2.1 FAME Extraction and Data Analysis . 183
12.2.2 PLFA Analysis . 185
12.2.3 Advantages and Limitations of Fatty Acid Patterns . 188
12.3 Denaturing Gradient Gel Electrophoresis . 189
12.3.1 DNA Extraction from Soil . 189
12.3.2 Polymerase Chain Reaction . 190
12.3.3 DGGE Procedures . 192
12.4 Conclusions . 196
References . ! 97
13 Measurement of Microbial Biomass and Activity in Soil . 201
Z. Solaiman
13.1 Introduction . 201
13.2 Protocols for Microbial Biomass Determination . 202
13.2.1 Chloroform Fumigation-Extraction Method
for Microbial Biomass C and N . 202
13.2.2 Hexanol Extraction Method for Microbial P . 205
13.3 Protocol for Total Microbial Activity Determination . 207
13.3.1 Equipment . 207
Contents
XVII
13.3.2 Reagents . 207
13.3.3 Protocol for Extraction . 208
13.4 Protocol for Soil Dehydrogenase Enzyme Analysis . 208
13.4.1 Equipment . 209
13.4.2 Reagents . 209
13.4.3 Protocol for Extraction . 209
13.5 Conclusions . 209
References . 210
14 Immuno-Technology for the Localization of Acid Phosphatase
Using Native Gel Bands in Piriformospora indica and Other Soil
Microorganisms . 213
R. Malla, U. Pokharel, R. Prasad, R. Oelmüller, and A. Varma
14.1 Introduction . 213
14.1.1 Taxonomic Status . 213
14.1.2 Phosphatases . 214
14.2 Immunotechnology for the Detection and Localization
of Acid Phosphatase in P. indica . 216
14.2.1 Extraction of Protein and Enzyme Assay . 216
14.2.2 Purification of Protein by Column Chromatography . 217
14.2.3 Purification of Protein by Ion Exchange
Chromatography. 217
14.2.4 Native Polyacrylamide Gel Electrophoresis . 219
14.2.5 Detection of Enzyme in Native PAGE . 220
14.2.6 Isolation of Acid Phosphatase for Raising Antibody . 222
14.2.7 Production of Antibodies using Acid Phosphatase
in Native Gel . 222
14.2.8 Antiserum Preparation . 224
14.2.9 Purification of Immunoglobulin from Serum . 225
14.2.10 Western Blot . 226
14.2.11 Immuno-Fluorescence . 228
14.2.12 Localization of ACPase by Immunogold Technique . 228
14.3 Troubleshooting . 232
14.4 Conclusions . 232
References . 232
15 Use of Short Oligonucleotide Primers in Random Amplified
Polymorphic DNA Techniques for Species Identification . 237
R. Malla and A. Varma
15.1 Introduction . 237
15.2 Polymorphism between Piriformospora indica and Sebacina
vermifera, Members of the Order Sebacinales . 239
15.3 General Protocol for RAPD Technique
to Show Polymorphism . 241
15.3.1 Experimental Procedures . 242
XVIII
Contents
15.4 Troubleshooting . 244
15.5 Conclusions . 244
References . 245
16 Co-Cultivation with Sebacinales . 247
A.C. Kharkwal, R. Prasad, H. Kharkwal, A. Das,
K. Bhatnagar, I. Sherameti, R. Oelmiiller, and A. Varma
16.1 Introduction . 247
16.2 Sebacinaceae - Novel Fungi . 248
16.3 Host Spectrum . 249
16.4 Functions of the Sebacinaceae . 251
16.5 Eco-Functional Identity . 252
16.6 Axenic Co-Cultivation of Sebacinaceae . 254
16.6.1 Procedure . 254
16.6.2 Protocol . 256
16.7 Media Compositions . 256
16.8 Seed Surface Sterilization and Germination . 261
16.8.1 Protocol for Seed Surface Sterilization . 262
16.8.2 Inoculum Placement in the Pots . 262
16.8.3 Results . 262
16.9 Comparative Study on Plant Growth with Treated
Endosymbionts . 264
16.10 In Vivo Co-Cultivation of Sebacinales . 264
16.11 Conclusions. 266
References . 267
17 Quantitative Histochemistry: a Forgotten Tool
with New Applications . 271
R. Hampp and S. Haag
17.1 Introduction . 271
17.2 Sample Preparation and Handling . 272
17.3 Microphotometry . 274
17.4 Biochemical Analysis: Real Time Microassays . 276
17.5 Spatial Resolution of Basic Steps
of Fungal Trehalose Metabolism in Symbiosis . 277
References . 279
18 Ion Cyclotron Resonance Fourier Transform Mass Spectrometry
for Non-Targeted Metabolomics of Molecular Interactions
in the Rhizosphere . 281
P. Schmitt-Kopplin, N. Hertkorn, M. Frommberger,
M. Ludo, M. Englmann, A. Fekete, and I. Gebefugi
18.1 Introduction . 281
18.2 The Chemical Biology Approach . 282
Contents
XIX
18.3 Complementary Analytical Approaches . 283
18.3.1 Targeted Analysis . 284
18.3.2 Metabolite Profiling . 285
18.3.3 Non-Targeted Analysis . 285
18.4 Resolving Structural Information from Molecular Complexity
with ICR-FT/MS . 286
18.4.1 Top-Down Approach: From ICR-FT/MS-Profiling
Analysis to Structural Hypothesis . 288
18.4.2 Complementary Analytical Tools . 290
18.4.3 Bottom-Up Approach: From Hypothesis-Driven
Experiments Upwards to ICR-FT/MS . 290
18.5 Conclusion . 292
References . 292
19 Application of Terminal-Restriction Fragment Length
Polymorphism for MolecularAnalysis of Soil Bacterial
Communities . 295
A. Mengoni, E. Giuntini, and M. Bazzicalupo
19.1 Introduction . 295
19.2 A General Protocol for Taxonomic T-RFLP Profiling of Soil
Bacterial Communities . 297
19.2.1 Materials . 297
19.2.2 Experimental Procedure . 298
19.2.3 Troubleshooting . 299
19.3 Standardization of T-RFLP Profiles . 299
19.4 Other Applications of T-RFLP to Soil Bacterial Communities 301
19.5 Conclusions . 302
References . 302
20 Molecular Symbiotic Analysis Between Arabiopsis thaliana
and Piriformospora indica . 307
B. Shahollari, K. Bhatnagar, I. Sherameti, A. Varma, and R. Oelmüller
20.1 Introduction . 307
20.2 Beneficial Interaction Between Plants and Fungi:
Piriformospora indica and Arabidopsis thaliana
as a Model System . 308
20.3 Co-Cultivation of P. indica and Arabidopsis under Standardized
Growth Conditions . 309
20.4 Map-Based Cloning of a Mutated Gene . 312
20.5 Rapid DNA Extraction . 313
20.6 Confirmation of a Mutated Phenotype of an EMS Mutant
by the Analysis of an Independent T-DNA Insertion Line . 313
20.7 Differntial Display to Identify Genes which are Regulated
in Response to P. indica . 314
20.8 Activation Tagged Lines . 315
XX
Contents
20.9 Identification of Biochemical Pathways
in A. thaliana which are Regulated by P. indica . 317
References . 317
21 Biophysical Phenomics Reveals Functional Building Blocks
of Plants Systems Biology: a Case Study for the Evaluation
of the Impact of Mycorrhization with Piriformospora indica . 319
R.}, Strasser, M. Tsimilli-Michael, D. Dangre, and M. Rai
21.1 Introduction . 319
21.2 Biophysical Phenomics of the Fast Fluorescence Rise O-J-I-P 320
21.2.1 The Energy Cascade in the Photosynthetic Apparatus 320
21.2.2 Microstates - Functional Building Blocks
of Photosynthesis . 320
21.2.3 Measuring Fluorescence Transients with PEA,
Handy-PEA and FIM- Fluorimeters . 322
21.2.4 How Fluorescence Kinetics Provide an Insight
to the Microstates - Functional Blocks of PSII . 323
21.3 Case Study . 332
21.3.1 Mycorrhization and the Advantages of Piriformospora
indica, an Emerging Growth Booster . 332
21.3.2 Phenomics of the O-J-I-P Fluorescence Transient
for the Study of Cadmium Stress on Chick Peas
(Cicer arietinum L. Chafa variety) With and Without
Symbiosis With Glomus mosseae, G. caledonium
and Piriformospora indica . 333
21.3.3 Correlation of Physiological with Biophysical
Parameters . 337
21.4 Conclusions . 338
References . 338
22 Analysis of the Plant Protective Potential of the Root Endophytic
Fungus Piriformospora indica in Cereals . 343
F. Waller, B. Achatz, and K.-H. Kogel
22.1 Introduction . 343
22.2 Plant Responses and Resistance to Pathogens . 344
22.2.1 Local Reactions . 344
22.2.2 Systemic Reactions and Resistance in Cereals . 344
22.2.3 Beneficial Microbial Endophytes Protecting Cereals
from Pathogens . 345
22.3 Interaction of P indica with Cereals . 345
22.3.1 P. indica Colonizes Root Cortical Cells in Barley. 346
22.3.2 P. indica Enhances Biomass and Yield in Barley . 346
22.4 Approaches to Study the Mechanism
of P indica-Induced Pathogen Resistance. 347
Contents
XXI
22.4.1 P indica Induces Disease Resistance Against Root
Pathogens . 347
22.4.2 P. indica Induces Systemic Disease Resistance . 348
22.4.3 Assessment of the Antioxidant Capacity of P. indica-
Infested Roots . 350
22.4.4 Gene Expression Induced by P. indica in Barley Leaves 351
22.5 Conclusions . 351
References . 352
23 Members of Sebacinales Confer Resistance Against Heavy Metal
Stress in Plants . 355
N. Hahn, A. Varma, R. Oelmüller, and I. Sherameti
23.1 Introduction . 355
23.2 Scientific Background . 355
23.3 Differential Display to Understand Cd2+ Resistance
Mediated by Endophytic Fungi . 357
23.4 Studies on Protein Level . 357
23.4.1 Two-Dimensional Gel Electrophoresis,
Preparation of Proteins . 359
23.4.2 Mass Spectrometry, Preparation of Samples
by Tryptic Digestion . 359
References . 360
24 Screening of Plant Growth-Promoting Rhizobacteria . 363
C.S. Nautiyal and S.M. DasGupta
24.1 Introduction . 363
24.2 Candidature for Being a Rhizobacteria . 364
24.3 Screening Methods . 365
24.3.1 Criteria for Screening . 365
24.3.2 Selection of Screening Methods . 365
24.3.3 Classic Methods . 366
24.3.4 Modern Methods . 368
24.3.5 Molecular Methods . 370
24.4 Metagenomics . 371
24.5 Tracking of GEMs . 372
24.6 Conclusions . 372
References . 373
25 Research Methods in Arbuscular Mycorrhizal Fungi . 377
A. Gaur and A. Varma
25.1 Introduction . 377
25.2 Assessment of AM Fungal Propagules in Soil . 378
25.2.1 Soil Sampling. 378
XXII
Contents
25.2.2 Spore Extraction . 378
25.2.3 Quantification of Spore Numbers . 379
25.2.4 Infectivity Assays . 379
25.2.5 Identification of AM Fungi . 380
25.2.6 Use of Fatty Acids for Identification of AM Fungi. 381
25.3 Quantification of AM Fungal Root Colonization in Root . 382
25.3.1 Clearing and Staining Roots . 382
25.3.2 Modifications of Staining Procedure . 383
25.3.3 Measurement of Root Colonization by AM Fungi . 384
25.4 Extraction and Quantification of Extra-Radical Mycelium
of AM Fungi in Soils . 384
25.5 Assessment of Growth Response of Effective Isolates . 385
25.6 Inoculum Production of AM Fungi . 386
25.6.1 On-Farm Production of AM Fungi . 386
25.6.2 Traditional Culture Methods . 387
25.6.3 AM Fungal Culture Using Aeroponic and Hydroponic
Culture . 388
25.6.4 Monoaxenic Culture of AM Fungi . 389
25.6.5 Storage of AM Fungal Inoculum . 390
25.7 Conclusions . 390
References . 391
26 Field Trials of Bioinoculants . 397
I. Orta$ and A. Varma
26.1 Introduction . 397
26.2 Effect of Mycorrhizal Infection on Nutrient Uptake . 398
26.3 Effect of Soil Fumigation and Mycorrhizal
Inoculation on Plant Growth Under Field Conditions . 399
26.4 Effect of Mycorrhizal Inoculation on Plant Growth
and Nutrient Uptake under Non-Sterile Field Conditions. 403
26.5 Soil and Crop Management System . 408
26.6 Inoculation Techniques . 409
26.7 Conclusion . 411
References . 412
Subject Index
415 |
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| genre | (DE-588)4143413-4 Aufsatzsammlung gnd-content |
| genre_facet | Aufsatzsammlung |
| id | DE-604.BV023031223 |
| illustrated | Illustrated |
| index_date | 2024-07-02T19:17:05Z |
| indexdate | 2024-07-09T21:09:24Z |
| institution | BVB |
| isbn | 9783540708643 3540708642 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-016235104 |
| oclc_num | 255966765 |
| open_access_boolean | |
| owner | DE-703 DE-634 DE-11 |
| owner_facet | DE-703 DE-634 DE-11 |
| physical | XXVIII, 427 S. Ill., graph. Darst. 24 cm |
| publishDate | 2007 |
| publishDateSearch | 2007 |
| publishDateSort | 2007 |
| publisher | Springer |
| record_format | marc |
| series | Soil biology |
| series2 | Soil biology |
| spelling | Advanced techniques in soil microbiology Ajit Varma ... (ed.) Berlin ; Heidelberg ; New York Springer 2007 XXVIII, 427 S. Ill., graph. Darst. 24 cm txt rdacontent n rdamedia nc rdacarrier Soil biology 11 Literaturangaben Bodenmikrobiologie - Aufsatzsammlung Soil microbiology Bodenmikrobiologie (DE-588)4659394-9 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Bodenmikrobiologie (DE-588)4659394-9 s DE-604 Varma, Ajit 1939- Sonstige (DE-588)120066718 oth Soil biology 11 (DE-604)BV019353150 11 KOBV Fremddatenuebernahme application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016235104&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Advanced techniques in soil microbiology Soil biology Bodenmikrobiologie - Aufsatzsammlung Soil microbiology Bodenmikrobiologie (DE-588)4659394-9 gnd |
| subject_GND | (DE-588)4659394-9 (DE-588)4143413-4 |
| title | Advanced techniques in soil microbiology |
| title_auth | Advanced techniques in soil microbiology |
| title_exact_search | Advanced techniques in soil microbiology |
| title_exact_search_txtP | Advanced techniques in soil microbiology |
| title_full | Advanced techniques in soil microbiology Ajit Varma ... (ed.) |
| title_fullStr | Advanced techniques in soil microbiology Ajit Varma ... (ed.) |
| title_full_unstemmed | Advanced techniques in soil microbiology Ajit Varma ... (ed.) |
| title_short | Advanced techniques in soil microbiology |
| title_sort | advanced techniques in soil microbiology |
| topic | Bodenmikrobiologie - Aufsatzsammlung Soil microbiology Bodenmikrobiologie (DE-588)4659394-9 gnd |
| topic_facet | Bodenmikrobiologie - Aufsatzsammlung Soil microbiology Bodenmikrobiologie Aufsatzsammlung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016235104&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV019353150 |
| work_keys_str_mv | AT varmaajit advancedtechniquesinsoilmicrobiology |