In Chloroplast Biotechnology: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used in chloroplast molecular biology. Chapters focus on essential background information, applications in tobacco and protocols for plastid transformation in crops and Chlamydomonas and Bryophytes. Written in the highly 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 laboratory protocols and key tips on troubleshooting and avoidance of known pitfalls.
Authoritative and practical, Chloroplast Biotechnology: Methods and Protocols seek to aid scientists who study chloroplast molecular biology as well as those interested in applications in agriculture, industrial biotechnology and healthcare.
Part I: Background
1. The Plastid Genomes of Flowering Plants
Tracey A. Ruhlman and Robert K. Jansen
2. Next Generation Technologies to Determine Plastid Genome Sequences
Robert J. Henry, Nicole Rice, Mark Edwards and Catherine J. Nock
3. Plastid Gene Transcription: Promoters and RNA Polymerases
Jennifer Ortelt and Gerhard Link
4. Plastid mRNA Translation
Masahiro Sugiura
5. Engineering Chloroplasts for High-Level Foreign Protein Expression
Ralph Bock
6. Excision of Plastid Marker Genes Using Directly Repeated DNA Sequences
Elisabeth A. Mudd, Panagiotis Madesis, Elena Martin Avila and Anil Day
7. Fluorescent Labeling and Confocal Microscopic Imaging of Chloroplasts and Non-Green Plastids
Maureen R. Hanson and Amirali Sattarzadeh
Part II: Nicotiana Tabacum the Model Species of Chloroplast Biotechnology
8. Plastid Transformation in Nicotiana tabacum and Nicotiana sylvestris by Biolistic DNA Delivery to Leaves
Pal Maliga and Tarinee Tungsuchat Huang
9. Nicotiana tabacum – PEG-mediated Plastid Transformation
Areli Herrera Díaz and Hans-Ulrich Koop
10. Plastid Transformation of Tobacco Suspension Cell Cultures
Jeffrey M. Staub
11. Tryptophan and Indole Analog Mediated Plastid Transformation
Pierluigi Barone, Xing-Hai Zhang and Jack M. Widholm
12. Plastid Marker Gene Excision in Greenhouse-Grown Tobacco by Agrobacterium-Delivered Cre Recombinase
Tarinee Tungsuchat-Huang and Pal Maliga
13. Determination of the Half-Life of Chloroplast Transcripts in Tobacco Leaves
Sithichoke Tangphatsornruang and John C. Gray
14. Quantification of Organellar DNA and RNA Using Real-Time PCR
Andreas Weihe
15. Plastid Transformation for Rubisco Engineering and Protocols for Assessing Expression
Spencer M. Whitney and Robert E. Sharwood
Part III: Crop Specific Plastid Transformation Protocols
16. Plastid Transformation in Tomato
Stephanie Ruf and Ralph Bock
17. Stable Plastid Transformation of Petunia
Elena Martin Avila and Anil Day
18. Plastid Transformation in Potato - Solanum tuberosum
Vladimir T. Valkov, Daniela Gargano, Nunzia Scotti, and Teodoro Cardi
19. Plastid Transformation in Eggplant
Kailash C. Bansal and Ajay K. Singh
20. Plastid Transformation in Lettuce (
Lactuca sativa L.) by Polyethylene Glycol Treatment of Protoplasts
Cilia L.C. Lelivelt, Kees M.P. van Dun, C. Bastiaan de Snoo, Matthew S. McCabe, Bridget V. Hogg, and Jacqueline M. Nugent
21. Plastid Transformation in Lettuce (Lactuca sativa L) by Biolistic DNA Delivery
Tracey A. Ruhlman
22. Plastid Transformation in Soybean
Manuel Dubald, Ghislaine Tissot and Bernard Pelissier
23. Plastid Transformation in Cabbage (Brassica oleracea L. var. capitata L.) by the Biolistic Process
Menq-Jiau Tseng, Ming-Te Yang, Wan-Ru Chu, and Cheng-Wei Liu
24. Plastid Transformation in Sugar Beet – Beta vulgaris
Francesca De Marchis and Michele Bellucci
25. Integration and Expression of gfp in the Plastid of Medicago sativa L.
Shaochen Xing, Zhengyi Wei, Yunpeng Wang, Yanzhi Liu, Chunjing Lin
Part IV: Plastid Transformation in Algae
26. Rapid Screening for the Robust Expression of Recombinant Proteins in Algal Plastids
Daniel Barrera, Javier Gimpel and Stephen Mayfield
27. A Simple, Low Cost Method for Chloroplast Transformation of the Green Alga Chlamydomonas reinhardtii
Chloe Economou, Thanyanan Wannathong, Joanna Szaub and Saul Purton
28. Tools for Regulated Gene Expression in the Chloroplast of Chlamydomonas
Jean-David Rochaix, Raymond Surzycki and Silvia Ramundo
Part V : Plastid Transformation in Bryophyte
29. Plastid Transformation in Physcomitrella patens
Mamoru Sugita
30. Plastid Transformation of Sporelings and Suspension-cultured Cells from the Liverwort Marchantia polymorpha L.
Shota Chiyoda, Katsuyuki T. Yamato, and Takayuki Kohchi
