Phytoplasmas are a group of bacteria that are capable of multiplying and causing severe diseases in a wide range of plant species and are transported between plants by insect vectors in which the bacteria can also multiply. Phytoplasmas can cause serious and devastating problems for crop plants, especially in developing countries.
In Phytoplasmas: Methods and Protocols experts in the field cover topics pertinent to scientists intending to develop a research program on phytoplasmas. The volume opens with an introductory chapter on the importance of phytoplasma diseases, followed by methods for detection and diagnosis as well as techniques for separating and classifying the phytoplasmas into their different taxonomic groups and subgroups. The final chapters cover methods for separating phytoplasma genomic and plasmid DNA from plant DNA for whole genome sequencing.
Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Phytoplasmas: Methods and Protocols serves as an ideal guide to plant pathologists and molecular biologists aiming to set up a diagnostics facility for identifying the presence of these pathogens in plants or their insect vectors.
1. The Phytoplasmas - An Introduction Matt Dickinson, Melanie Tuffen, and Jennifer Hodgetts
2. Techniques for the Maintenance and Propagation of Phytoplasmas in Glasshouse Collections of Catharanthus roseus Jennifer Hodgetts, David Crossley, and Matt Dickinson
3. Micropropagation and Maintenance of Phytoplasmas in Tissue Culture Assunta Bertaccini, Samanta Paltrinieri, Marta Martini, Mara Tedeschi, and Nicoletta Contaldo
4. Dodder Transmission of Phytoplasmas Jaroslava Pribylova and Josef Spak
5. Insect Maintenance and Transmission Heather Kingdom
6. Capturing Insect Vectors of Phytoplasmas Phyllis Weintraub and Jurgen Gross
7. Insect Vector Transmission Assays Domenico Bosco and Rosemarie Tedeschi
8. Molecular Identification of Phytoplasma Vector Species Sabrina Bertin and Domenico Bosco
9. Dienes' Staining and Light Microscopy for Phytoplasma Visualization Rita Musetti
10. DAPI Staining and Fluorescence Microscopy Techniques for Phytoplasmas Nancy S. Andrade and Nolberto S. Arismendi
11. Visualisation of Phytoplasmas using Electron Microscopy B. Jean Devonshire
12. Automated DNA Extraction for Large Numbers of Plant Samples Natasa Mehle, Petra Nikolic, Matevz Rupar, Jana Boben, Maja Ravnikar, and Marina Dermastia
13. DNA Extraction from Arborescent Monocots and How To Deal With Other Challenging Hosts Nigel A. Harrison, Robert E. Davis, and Ericka E. Helmick
14. Nested PCR and RFLP Analysis Based on the 16S rRNA Gene Bojan Duduk, Samantha Paltrinieri, Ing Ming Lee, and Assunta Bertaccini
15. PCR and RFLP Analyses Based on the Ribosomal Protein Operon Marta Martini and Ing-Ming Lee
16. Tuf and secY PCR Amplification and Genotyping of Phytoplasmas Xavier Foissac, Jean-Luc Danet, Sylvie Malembic-Maher, Pascal Salar, Dana Safarova, Pavla Valova, and Milan Navratil
17. PCR Analysis of Phytoplasmas Based on the secA Gene Matt Dickinson and Jennifer Hodgetts
18. Single-Strand Conformation Polymorphism Analysis for Differentiating Phytoplasma Strains Martina Seruga Music and Dijana Skoric
19. Microarrays for Universal Detection and Identification of Phytoplasmas Mogens Nicolaisen, Henriette Nyskjold, and Assunta Bertaccine
20. T-RFLP for Detection and Identification of Phytoplasmas in Plants Jennifer Hodgetts and Matt Dickinson
21. Real-Time PCR for Universal Phytoplasma Detection and Quantification Nynne Meyn Christensen, Henriette Nyskjold, and Mogens Nicolaisen
22. A Real-Time PCR Detection System for the Bois Noir and Flavescence Doree Phytoplasmas and Quantification of the Target DNA Natasa Mehle, Nina Prezelj, Matjaz Hren, Jana Boben, Kristina Gruden, Maja Ravnikar, and Marina Dermastia
23. Real-Time PCR for Specific Detection of Three Phytoplasmas from the Apple Proliferation Group Natasa Mehle, Petra Nikolic, Kristina Gruden, Maja Ravnikar, and Marina Dermastia
24. Reverse Transcription-PCR for Phytoplasma Detection Utilizing Crude Sap Extractions Paola Margaria and Sabrina Palmano
25. In-Field Diagnostics Using Loop-Mediated Isothermal Amplification (LAMP) Jenny Tomlinson
26. DNA Bar-Coding for Phytoplasma Identification Olga Makarova, Nicoletta Contaldo, Samanta Paltrinieri, Assunta Bertaccini, Henriette Nyskjold, and Mogens Nicolaisen
27. Phylogenetic Analyses of Phytoplasmas Based on Whole-Genome Comparison Hiromi Nishida
28. The iPhyClassifier, An Interactive Online Tool for Phytoplasma Classification and Taxonomic Assignment Yan Zhao, Wei Wei, Ing-Ming Lee, Jonathan Shao, Xiaobing Suo, and Robert E. Davis
29. Phytoplasma Proteomic Analysis Xianling Ji and Yingping Gai
30. Preparation of Phytoplasma Membrane Recombinant Proteins Luciana Galetto, Majid Siampour, and Cristina Marzachi
31. Phytoplasma Plasmid DNA Extraction Mark T. Andersen and Lia W. Liefting
32. Caesium Chloride-Bisbenzimide Gradients for Separation of Phytoplasma and Plant DNA Lucy T.T. Tran-Nguyen and Bernd Schneider
33. Pulsed-Field Gel Elecrophoresis for Isolation of Full-Length Phytoplasma Chromosomes From Plants Carmine Marcone
34. Mapping the Phytoplasma Chromosome Sylvie Malembic-Maher and Patricia Carle