laboratory setup of plant tissue culture lab

1. LABORATORY SETUP OF PTC LAB submitted by , Barsha Mahanandia Reg No - 2022AGPB124 Dept of Botany College of Agriculture VNMKV Parbhani

2. INTRODUCTION TO PTC Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues, or organs under sterile conditions on a nutrient culture medium of known composition. It is widely used to produce clones of a plant in a method known as micropropagation.

3.  INFRASTRUCTURE REQUIREMENTS 1 - Clean, well-ventilated room 2 - Controlled temperature and humidity 3 - Dedicated workbenches 4 - Proper lighting conditions

4. EQUIPMENT 1 - Autoclave 2 -Laminar flow hood 3 - UV sterilizer 4 - Microscopes 5 - pH meter 6 - Balances 7 - Pipettes 8 - Plant growth chambers or incubators

5. CONSUMABLES AND REAGENTS 1 - Culture media (MS medium, B5 medium, etc.) 2 - Agar, sugar, vitamins, amino acids 3 - Sterilization chemicals 4 - Supplements

6. STERILIZATION PROTOCOL 1 - Importance of sterilization < Sterilization of explants is a key step in any plant tissue culture work, as the removal of all microorganisms, including bacteria and fungi, is essential to attain successful initiation, growth and development of the cultured tissues in vitro, which otherwise would be overwhelmed by the contaminants > 2 -Procedures for sterilizing equipment, media, and plant materia <lMany laboratories use autoclaves for sterilizing media which works on the principle of moist heat. Moist heat sterilization is a procedure in which heated, high-pressure steam is used to sterilize an object (at 121-degree °C, 1.03 bar pressure for 15-20 minutes). . 3 - Maintaining sterility to prevent contamination

7.  WORKFLOWS 1 - Initiating cultures <The tissue is initiated in the culture at this time. The tissue of interest is acquired, introduced, and sterilized to avoid contamination. Multiplication Phase: The sterile explant is placed in a medium containing growth regulators and required nutrients. > 2 - Subculturing <This is mainly done when plants have completely outgrown their previous culture jar or the media is exhausted (or both factors). In such cases, plant tissues are divided into small sections are transferred to fresh media. The number of times the subculturing process is done, it's known as the “propagation cycle > 3 - Maintaining cultures 4 -Aseptic techniques and documentation < Always wipe your hands and your work area with 70% ethanol. Wipe the outside of the containers, flasks, plates, and dishes with 70% ethanol before placing them in the cell culture hood. Avoid pouring media and reagents directly from bottles or flasks. >

8. QUALITY CONTROL 1 - Monitoring for contamination < Regular monitoring and visual inspection of plant tissue cultures are essential to detect signs of contamination. Visual and microscopic examinations of cultures can help determine the presence of microbial contamination > 2 - Techniques for identifying and managing contamination < The authors suggested several techniques to prevent contamination. These include maintaining a clean and sterile environment, proper use of personal protective equipment, aseptic techniques for handling cultures and equipment, regular monitoring of cultures for contamination, and use of sterile media and equipment > 3 - Genetic stability < Transformation is the step in the genetic engineering process where a new gene (transgene) is inserted into a single plant cell. The transformation step of producing a genetically engineered (GE) crop plant is a process controlled by poorly understood mechanisms. >

9.  SAFETY MEASURES 1 - Personal protective equipment (PPE) 2 - Handling hazardous chemicals and biological materials 3 - Disposal procedures

10.  CONCLUSION In conclusion, the establishment of a plant tissue culture laboratory is fundamental for advancing various aspects of plant science, agriculture, and biotechnology. Through meticulous attention to infrastructure, equipment, protocols, and safety measures, we create an environment conducive to the precise manipulation and propagation of plant cells and tissues. By providing controlled conditions of temperature, humidity, and lighting, coupled with sterile workspaces and equipment, we ensure the success of tissue culture techniques. This enables us to initiate cultures, perform subculturing, and maintain genetic stability, thereby harnessing the potential of plant cells for diverse applications. Moreover, the adherence to strict sterilization protocols and quality control measures is paramount. Vigilance in monitoring for contamination and ensuring genetic fidelity guarantees the reliability and reproducibility of our results. In parallel, prioritizing safety through the use of personal protective equipment and proper handling of hazardous materials safeguards the well-being of laboratory personnel and the integrity of our experiments. Ultimately, the establishment of a plant tissue culture laboratory represents a commitment to innovation, sustainability, and the advancement of plant science. By harnessing the power of tissue culture techniques, we can contribute to agricultural productivity, genetic improvement, and the conservation of plant biodiversity, thus paving the way for a greener, healthier future. Thank you for joining us on this journey into the world of plant tissue culture. Together, let us continue to explore and unlock the immense potential that lies within the cells of plants.

11. REFRENCES  1 -George, E. F., et al. (2008). "Plant propagation by tissue culture."  2 - Murashige, T., & Skoog, F. (1962). "A revised medium for rapid growth and bio assays with tobacco tissue cultures." 3 - Google

12. THANK YOU