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The fundamental concept of totipotency that a single cell has the potential to grow into a whole mature plant- is at the heart of plant tissue culture. “ Practice patience...

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Tissue Culture Technology
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The fundamental concept of totipotency that a single cell has the potential to grow into a whole mature plant- is at the heart of plant tissue culture. “ Practice patience and avoid rushing …take a dreep breath, slow down, and remember that patience is your ally.” This speaks to the meticulous, detail oriented work involved in tissue culture, where sterility and precision are critical. S.Y.B.Sc. BOT-242-MNT and BOT-245 MNP Tissue Culture Technology one of such course will make the students understanding about potential of plant cells and tissues when maintained at laboratory conditions with hormones, nutrients etc. under human control.

Tissue culture is the process that exploits the ability of plant cells to dedifferentiate and then redifferentiate to form a whole plant. Plant tissue culture is a key aspect of modern biotechnology, offering potential for progress and innovation in agriculture as well in forestry, industrial production of variety of plant secondary metabolites. The content of this course introduces variety of concepts about plant tissue culture, followed by laboratory setup and techniques, plant tissue culture media, concept and techniques of micropropagation, applied aspects of plant tissue culture techniques like callus culture, cell suspension culture, protoplast fusion, haploid production of plants also.

The germplasm conservation is a key process in further management of plant cells by cryopreservation. We authors have taken serious efforts to present the course matter in most simple way so that our students and our teachers will appreciate the style of presentation. The course matter contains variety of concepts, so we have tried our best to express this with our sincere efforts. We authors are thankful to our publisher Hon. Shri. Rangrao Patil, who has taken meticulous efforts to publish series of all titles.

We are also thankful to all the staff of Prashant Publications, Jalgoan for doing sincere efforts to present the alphabetical in a presentable form called Tissue Culture Technology. We all authors are thankful to several Publications, authors, organisations from where we have taken theoretical matter for our presentations. We are also thankful to our students and teacher friends which are our source of support and inspiration. We always take criticisms, suggestions and comments for further improvement of our books in future.

BOT 242 (MN) – TISSUE CULTURE TECHONOLOGY
1. Introduction to Plant Tissue Culture………………………………………………. 9
1.1. Introduction and brief history of PTC.
1.2. Definitions and Concepts
1.3 Solution concentration
1.4 Nutrient medium and Related concepts
1.5 Organogenesis and other concepts
1.6. Exercise
2. Laboratory Setup and Requirements in PTC………………………………… 37
2.1. Introduction.
2.2. Layout and Design of plant tissue culture laboratory (PTC)
2.3. Basic Requirements, Chemicals in PTC.
2.4. Glassware and plastic ware in PTC.
2.5. Equipment/Instrumentsin PTC.
2.6. Miscellaneouscomponents in PTC.
2.7 Exercise
3. Aseptic / Sterilization Techniques…………………………………………………. 62
3.1. Introduction, definition and concept.
3.2. Methods of sterilization
3.3 Physical methods
3.4 Chemical methods: of sterilization.
3.5 Exercise
4. Tissue Culture Media…………………………………………………………………… 70
4.1. Introduction, definition, and concept of medium.
4.2. Role of essential nutrients in plant growth and development.
4.3. Types of culture media: MS, B5, White’s and Nitsch medium.
4.4. Composition of MS nutrient culture media-
4.5. Media preparation
4.6. Exercise
5. Micropropagation………………………………………………………………………… 90
5.1 Introduction, Definition and Concept
5.2 Stages of Micropropagation
5.3 Organogenesis
5.4 Commercial Micropropagation of Banana and Pomegranate
5.5 Advantages and limitations of Micropropagation
5.6 Exercise

6. Applied Plant Tissue Culture Techniques……………………………………… 99
6.1 Introduction
6.2 Callus culture:
6.3 Cell susupension culture
6.4 Somatic embryogenesis
6.5 Protoplast culture and fusion
6.6 Haploid culture
6.7 Embryo and ovule culture
6.8 Embryo Rescue
6.9 Exercise
7. Cryopreservation and Germplasm Conservation………………………… 130
7.1 Cryopreservation: Introduction and Definition
7.2 Methods of cryopreservation
7.3 Steps involved in cryopreservation
7.4 Application of cryopreservation
7.5 Germplasm Conservation
7.6 Exercise

BOT-245 (MNP)-PRACTICAL BASED ON BOT-242-MN
1. To study the basic layout of typical Plant Tissue Culture Laboratory and
various sterilization methods used in PTC using ICT tools……………………………… 140
2. To study the principle, working and applications of instruments/equipment/
glassware used in PTC laboratory………………………………………………………………… 145
3. Basics of laboratory mathematics, calculations for solution preparation……………… 158
4. To perform the procedure of stocks solution preparation for various components
of MS basal media and plant growth regulators (PGRs)………………………………….. 160
5. Preparation and Sterilization of MS Medium for Organogenesis and
Callogenesis……………………………………………………………………………………………… 166
6. Surface Sterilization and Inoculation of Leaf/Apical Meristem/Nodal Segment
for Organogenesis and Callogenesis…………………………………………………………….. 171
7. To study the steps involved in sub culturing of proliferated shoots and roots on
freshly prepared nutrient media…………………………………………………………………….. 173
8. To study the procedure involved in harding of rooted plants and its
acclimatization………………………………………………………………………………………….. 174
9. Micropropagation of commercially important plants:
Banana/Gerbera/Strawberry/Pomegranate…………………………………………………….. 176
10. Initiation of cell suspension culture and its growth measurement…………………….. 178
11. Visit to the plant tissue culture laboratory and preparation of visit report………….. 181