Studies on molecular biology of pathogens, infection process and disease resistance, have provided information essentially required to understand the vulnerable stages at which the pathogens can be tackled effectively and to adopt novel strategies to incorporate disease resistance genes from diverse sources and /or to induce resistance of cultivars with desirable agronomic attributes using biotic or abiotic agents. The nature of interaction between the gene products of the pathogen and plant appears to determine the outcome of the interaction resulting in either disease progression or suppression. Transgenic plants with engineered genes show promise for effective exploitation of this approach for practical application. Research efforts during the recent years to sequence the whole genomes of the pathogens and plants may lead to development of better ways of manipulating disease resistance mechanisms enabling the grower to achieve higher production levels and the consumer to enjoy safer food and agricultural products. Experimental protocols included in appropriate chapters will be useful for researchers and graduate students.
Author(s): P. Narayanasamy
Edition: 1
Publisher: Springer
Year: 2008
Language: English
Pages: 262
1402082428......Page 1
Contents......Page 6
Preface......Page 13
Acknowledgement......Page 15
1.1 Molecular Biology as a Research Tool......Page 16
1.2 Application of Molecular Techniques......Page 18
References......Page 21
2. Molecular Techniques for Detection of Microbial Pathogens......Page 22
2.1.1 Electrophoresis......Page 24
2.2 Detection of Microbial Pathogens by Immunoassays......Page 29
2.2.1 Viral Pathogens......Page 30
2.2.2 Bacterial Pathogens......Page 40
2.2.3 Fungal Pathogens......Page 44
2.3 Detection of Microbial Plant Pathogens by Nucleic Acid-Based Techniques......Page 47
2.3.1 Detection of Viral Pathogens......Page 49
2.3.2 Detection of Viroids......Page 72
2.3.3 Detection of Bacterial Pathogens......Page 75
2.3.4 Detection of Phytoplasmal Pathogens......Page 95
2.3.5 Detection of Fungal Pathogens......Page 100
Appendix 1: Electrophoretic Characterization of Strains of Bacterial Pathogen Xanthomonas campestris pv. vesicatoria (Xcv) (Bouzar et al. 1994)......Page 125
Appendix 2: Detection of Virus-Specific Protein in Infected Leaves by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) (Seifers et al. 1996, 2005)......Page 126
Appendix 3: Indirect ELISA for Assessing Titers of PABs and MABs Specific to Callalily chlorotic spot virus (CCSV) and Watermelon silver mottle virus (WSMoV) (Lin et al. 2005)......Page 127
Appendix 5: Detection of Potato virus Y (PVY) and Cucumber mosaic virus (CMV) in Tobacco by Immunostaining Technique (Ryang et al. 2004)......Page 128
Appendix 6: Detection of Citrus tristeza virus (CTV) by In Situ Immunoassay (ISIA) (Lin et al. 2000)......Page 129
Appendix 8: Detection of Bacterial Pathogens by Enzyme-Linked Immunosorbent Assay (ELISA) in Seeds (Lamka et al. 1991; Pataky et al. 2004)......Page 130
Appendix 9: Detection of Ultilago nuda Barley Seeds by DAS-ELISA (Eibel et al. 2005)......Page 131
Appendix 10: Detection of Plant Viruses by Reverse-Transcription-Polymerase Chain Reaction (RT-PCR) Assay (Huang et al. 2004; Spiegel et al. 2004)......Page 133
Appendix 11: Detection of Virus (Potato virus Y) by Reverse Transcription – DIAPOPS System (Nicolaisen et al. 2001)......Page 134
Appendix 12: Detection of Grape fan leaf virus (GFLV) in Nematode Vector Xiphinema index by RT-PCR (Finetti-Sialer and Ciancio 2005)......Page 136
Appendix 13: Detection of Potato virus Y by Reverse Transcription Loop-Mediated Isothermal Amplification DNA (Nie 2005)......Page 137
Appendix 14: Detection of Fruit Tree Viroids by a Rapid RT-PCR Test (Hassen et al. 2006)......Page 138
Appendix 15: Membrane BIO-PCR Technique for Detection of Bacterial Pathogen (Pseudomonas syringae pv. phaseolicola) (Schaad et al. 2007)......Page 140
Appendix 16: Detection of Bacterial Pathogens by DNA Array Technology (Fessehaie et al. 2003; Scholberg et al. 2005)......Page 141
Appendix 17: Extraction of Genomic DNA from Fungal Pathogens (Phytophthora spp.) (Lamour and Finley 2006)......Page 143
Appendix 18: Detection of Mycosphaerella graminicola in Wheat Using Reverse Transcription (RT)-PCR (Guo et al. 2005)......Page 144
References......Page 145
3. Molecular Variability of Microbial Plant Pathogens......Page 174
3.1 Molecular Basis of Variability of Fungal Pathogens......Page 175
3.1.1 Isozyme Variation......Page 176
3.1.2 Immunological Assay......Page 177
3.1.4 Restriction Fragment Length Polymorphism......Page 178
3.1.5 Polymerase Chain Reaction......Page 183
3.1.6 Random Amplified Polymorphic DNA Technique......Page 190
3.1.7 Amplified Fragment Length Polymorphism Technique......Page 194
3.1.9 Microsatellite Amplification......Page 198
3.1.10 Single-Strand Conformation Polymorphism Analysis......Page 199
3.2.1 Immunoassays......Page 200
3.2.2 Restriction Fragment Length Polymorphism......Page 201
3.2.3 Polymerase Chain Reaction......Page 202
3.2.4 Random Amplified Polymorphic DNA......Page 206
3.2.5 DNA–DNA Hybridization......Page 208
3.2.7 PCR-Based Suppression Subtractive Hybridization......Page 210
3.3.1 Immunological Techniques......Page 211
3.3.2 Nucleic Acid-Based Techniques......Page 215
3.4 Molecular Basis of Variability of Viroid Pathogens......Page 223
Appendix 1: Microsatellite-Primed (MP) Polymerase Chain Reaction for DNA Fingerprinting (Ma and Michailides 2005)......Page 224
Appendix 2: Amplified Fragment Length Polymorphism (AFLP) Analysis of Pythium spp. (Garzón et al. 2005)......Page 225
References......Page 226
A......Page 241
B......Page 243
C......Page 244
E......Page 246
F......Page 247
G......Page 248
H......Page 249
K......Page 250
M......Page 251
P......Page 252
Q......Page 253
S......Page 254
T......Page 255
Z......Page 257
D......Page 258
I......Page 259
P......Page 260
T......Page 261
Z......Page 262
