Plant tissue culture is a basic, fundamental science from the branch of plant biotechnology that helps in understanding the growth and development of plants at the cellular level. Mainly plant tissue culture is used for crop improvement either by developing new varieties through different techniques, such as somaclonal variation, protoplast fusion, etc., or genetic modification through transgenic technology i.e. the cells of the plants can be genetically altered and the modified cell will undergo for regeneration into whole plant with desirable characteristics through tissue culture technology. So, to develop transgenic crops with desirable traits, one should know thoroughly about tissue culture techniques and basics of molecular biology. This book deals with essential elements of Plant tissue culture techniques viz., Meristem culture, Haploid plant production, Somatic embryogenesis, Somoclonal variation, Protoplast fusion, Micropropagation, Secondary metabolite production and Germplasm conservation. In Molecular biology aspects, it covers Basics of nucleic acids, Gene structure, Central Dogma, Polymerase chain reaction, Sequencing and Blotting techniques, Recombinant technology, Cloning vectors, Plant transformation methods, Transgenic crops development for biotic, abiotic stress and quality improvement, DNA markers, DNA fingerprinting, Linkage and QTL analysis and Marker assisted selection. This book has been written in a very simple and crisp manner enabling easy understanding by the readers and students. This book also contains important objective type questions in each chapter at the end. This will be more useful for the students for their both University examination and competitive examination.
Dr. Sivaji Mathivanan, is presently working as a Teaching Assistant (Biotechnology) in Agriculture College & Research Institute (AC&RI), Vazhavachanur, Tiruvannamalai, one of the constituent college of Tamil Nadu Agricultural University (TNAU), Coimbatore. He has started his undergraduate programme B.Sc. Agriculture in Anbil Dharmalingam Agricultural College and Research Institute (ADAC&RI), Trichy, TNAU followed by Master Degree Programme M.Sc. Agri. in Plant Biotechnology in Kerala Agriculture University (KAU), Thrissur, Kerala through DBT-JNU Fellowship and Ph.D. in Biotechnology at AC&RI, Tamil Nadu Agricultural University, Coimbatore. He has qualified ICAR-SRF and ICAR-NET. He is having eight years working experience in both research and teaching. He has published 8 books, 35 national and internationalresearch papers, 20 book chapters in Springer, Apple academic press and other popular publishers and 25 popular articles in various aspects of Biotechnology. He has participated and presented 30 national and international seminar and symposium.
Shakespear Sundaresan, Currently doing M.Sc. Plant Biotechnology in the department of Molecular Biology and Biotechnology at ICAR-National Institute of Biotic Stress Management, Indian Agricultural Research Institute, Baronda, Raipur, Chhattisgarh. He has completed his Undergraduation from Agricultural College & Research Institute (AC&RI), Tamil Nadu Agricultural University, Vazhavachanur, Tiruvannamalai, Tamil Nadu. Driven by an inquisitive curiosity about biotechnology research, he went through the process of clearing ICAR-AIEEA for PG, and got ranking 9th in the wise category and 98th in the All India Ranking. As part of Post Graduate degree currently, he is doing research on “broad-spectrum disease resistance against bacterial leaf blight in cultivated and wild Oryza species”. He has participated in several Workshops, Training, National and International conferences and aware of current situations and recent advances in Biotechnology and Molecular Biology. He has published two research articles, two book chapters and several popular articles in various section of Biotechnology field.
Dr. K. Sakthivel is currently working as a Associate Professor in the Department of Genetics and Plant Breeding, Tamil Nadu Agricultural University – Agricultural College and Research Institute, Vazhavachanur. He has completed his Ph.D. from University of Agricultural Sciences, Dharwad and worked in national and central institutes/university. He possesses more than twenty years of research experience in various crops and in various research fields including Plant Breeding, Crop Molecular Breeding, TILLING, Agricultural Biotechnology, Developmental Physiology, etc. He has published more than 20 research papers in peer reviewed, high impact factor journals. He had released three crop varieties for cultivation by farmers. Currently he teaches Genetics, Agricultural Botany courses to the undergraduate students.
1. Plant Tissue Culture: Concepts, History and Scope ..................... 1
2. Components of Plant Tissue Culture, Media and Sterilization Techniques ..................................................................... 9
3. In vitro Plant Regeneration Methods ............................................. 23
4. Different Types of Culture Condition in Plant Tissue Culture .................................................................................... 35
5. Haploid Plant Production Through Androgenesis and Gynogenesis ............................................................................... 45
6. Micropropagation or in vitro Clonal Propagation of Banana and Ornamental Plants ..................................................... 57
7. National Certification System for Tissue Culture Raised Plants ...................................................................................... 71
8. Meristem Tip Culture (Virus Free Plants) and Anther Culture (Double Haploids) ................................................ 81
9. Protoplast Isolation and Fusion: Somaclonal Variation and Synthetic Seeds .......................................................................... 91
10. Secondary Metabolites Production and in vitro Germplasm Conservation ..............................................................111
11. Genome Organization in Prokaryotes and Eukaryotes ...........127
12. Central Dogma of Life and Structure of Nucleic Acids ...........139
13. DNA Replication, Genetic Code and DNA Transcription ......151
14. mRNA Translation and Protein Synthesis .................................171
15. Structure of a Gene, Regulation of Gene Expression and Operon Concept ..............................................................................183
16. Blotting Techniques, Polymerase Chain Reaction and DNA Sequencing Methods ............................................................197
17. DNA Manipulation Enzymes: Polymerase, Restriction Endonucleases and Ligases ...........................................................211
18. Different Types of Vectors: Plasmids, Phagemids, Cosmids and Bacterial Artificial Chromosome ..........................................225
19. Construction of Recombinant DNA Molecules, Bacterial Transformation, Direct and Indirect Gene Transfer Methods in Plants ...........................................................241
20. Tissue Specific Promoters, Selectable and Scorable Markers and Molecular Analysis of Transgenic Plants ...........................263
21. Transgenic Plants: Biotic and Abiotic Stress Resistance and Traits for Improved Quality ..................................................273
22. Detection of Genetically Modified Organisms, Their Regulations and Biosafety...................................................299
23. Molecular Markers, DNA Fingerprinting, Linkage and QTL Analysis ...................................................................................311
24. Principles, Methods and Applications of Marker Assisted Selection in Crop Improvement ...................................335
25. Applications of Plant Genomics and Genome Databases .......347
References .........................................................................................359