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Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            1 
 
      
Lesson Prepared Under MHRD project “Natio n al Mission on 
Education Through IC T” 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Laboratory Requirements in Plant Tissue Culture 
Lesson Developer: Dr. Ashima Khurana 
Department of Botany, Zakir Hussain Delhi College,  
University of Delhi and  
Dr. Rahul Kumar 
RTGR, Department of Plant Sciences, University of 
Hyderabad 
Lesson Reviewer: Dr Rita Sharma 
School of Life Sciences, Jawaharlal Nehru University 
Language Editor: Namrata Dhaka 
Department/College: Department of Genetics, University 
of Delhi  South Campus 
Lesson Editor: Dr Rama Sisodia, Fellow in Botany ILLL 
 
Page 2


Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            1 
 
      
Lesson Prepared Under MHRD project “Natio n al Mission on 
Education Through IC T” 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Laboratory Requirements in Plant Tissue Culture 
Lesson Developer: Dr. Ashima Khurana 
Department of Botany, Zakir Hussain Delhi College,  
University of Delhi and  
Dr. Rahul Kumar 
RTGR, Department of Plant Sciences, University of 
Hyderabad 
Lesson Reviewer: Dr Rita Sharma 
School of Life Sciences, Jawaharlal Nehru University 
Language Editor: Namrata Dhaka 
Department/College: Department of Genetics, University 
of Delhi  South Campus 
Lesson Editor: Dr Rama Sisodia, Fellow in Botany ILLL 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            2 
 
Learning Outcomes 
The chapter is aimed to enable the readers to learn about the following: 
? Basic facilities required to start a plant tissue culture laboratory.  
? Function of all these facilities and their importance in context of successful tissue 
culture practices. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 3


Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            1 
 
      
Lesson Prepared Under MHRD project “Natio n al Mission on 
Education Through IC T” 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Laboratory Requirements in Plant Tissue Culture 
Lesson Developer: Dr. Ashima Khurana 
Department of Botany, Zakir Hussain Delhi College,  
University of Delhi and  
Dr. Rahul Kumar 
RTGR, Department of Plant Sciences, University of 
Hyderabad 
Lesson Reviewer: Dr Rita Sharma 
School of Life Sciences, Jawaharlal Nehru University 
Language Editor: Namrata Dhaka 
Department/College: Department of Genetics, University 
of Delhi  South Campus 
Lesson Editor: Dr Rama Sisodia, Fellow in Botany ILLL 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            2 
 
Learning Outcomes 
The chapter is aimed to enable the readers to learn about the following: 
? Basic facilities required to start a plant tissue culture laboratory.  
? Function of all these facilities and their importance in context of successful tissue 
culture practices. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            3 
 
Chapter: Laboratory Requirements in Plant Tissue Culture 
Table of Contents  
? Introduction 
? Basic facilities required in a tissue culture laboratory 
? Washing area 
? Glassware/Plastic ware 
? Area for media preparation 
? Transfer area 
? Culture room and incubators 
? Greenhouse 
? Basic laboratory equipment and apparatus 
? Summary  
? Exercise/ Practice 
? Glossary 
? References/ Bibliography/ Further Reading 
 
 
 
 
 
 
 
 
 
 
Page 4


Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            1 
 
      
Lesson Prepared Under MHRD project “Natio n al Mission on 
Education Through IC T” 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Laboratory Requirements in Plant Tissue Culture 
Lesson Developer: Dr. Ashima Khurana 
Department of Botany, Zakir Hussain Delhi College,  
University of Delhi and  
Dr. Rahul Kumar 
RTGR, Department of Plant Sciences, University of 
Hyderabad 
Lesson Reviewer: Dr Rita Sharma 
School of Life Sciences, Jawaharlal Nehru University 
Language Editor: Namrata Dhaka 
Department/College: Department of Genetics, University 
of Delhi  South Campus 
Lesson Editor: Dr Rama Sisodia, Fellow in Botany ILLL 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            2 
 
Learning Outcomes 
The chapter is aimed to enable the readers to learn about the following: 
? Basic facilities required to start a plant tissue culture laboratory.  
? Function of all these facilities and their importance in context of successful tissue 
culture practices. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            3 
 
Chapter: Laboratory Requirements in Plant Tissue Culture 
Table of Contents  
? Introduction 
? Basic facilities required in a tissue culture laboratory 
? Washing area 
? Glassware/Plastic ware 
? Area for media preparation 
? Transfer area 
? Culture room and incubators 
? Greenhouse 
? Basic laboratory equipment and apparatus 
? Summary  
? Exercise/ Practice 
? Glossary 
? References/ Bibliography/ Further Reading 
 
 
 
 
 
 
 
 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            4 
 
Introduction 
Plant tissue culture techniques have played a pivotal role in addressing many questions of 
basic and applied fields of plant science. Initially, this technique was used for studying the 
roles of hormones in cytodifferentiation and organogenesis. It is now also used for the 
functional characterization of genes, elucidation of underlying regulatory mechanisms by 
developing genetically modified (transgenic) plants. Due to their intensive use in various 
fields of applied plant science, plant tissue culture techniques have also revolutionized the 
agriculture sector in modern times. For example, selected plants tissues/cells are cultured 
as suspended cells to produce plant products. Additionally, transgenic plants with enhanced 
agronomic traits such as high carotenoids (golden rice: Ye et al., 2000) or longer shelf life 
(Flavr Savr tomatoes: Redenbough et al., 1992) have been produced. Further, tissue culture 
methods also help in developing the somatic haploid embryos. Such somatic haploid 
embryos can subsequently be used for the production of homozygous plants. Hence, tissue 
culture techniques have played an important role in the progression of academic 
and applied plant science. 
The major applications of plant tissue culture techniques are: 
? The large scale production of ornamental plants. 
? Conservation of endangered plant species. 
? Production of plant-derived secondary metabolites and recombinant proteins on a large-
scale in liquid culture of plant cells in bioreactors. 
? Production of novel hybrids by either fusing protoplasts of distantly-related species or 
embryo-rescue technique. 
? Molecular, pharmacological and biochemical investigations of different aspects of plant 
growth and development such as in vitro flowering. 
? For the induction of polyploidy, by treating plants with of antimitotic agents such 
as colchicine. 
? To produce virus free plants from virus-infected stock, such as potatoes by culturing 
meristem/tip culture. 
? Production of genetically modified crops with improved agronomic traits. 
 
 
Page 5


Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            1 
 
      
Lesson Prepared Under MHRD project “Natio n al Mission on 
Education Through IC T” 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Laboratory Requirements in Plant Tissue Culture 
Lesson Developer: Dr. Ashima Khurana 
Department of Botany, Zakir Hussain Delhi College,  
University of Delhi and  
Dr. Rahul Kumar 
RTGR, Department of Plant Sciences, University of 
Hyderabad 
Lesson Reviewer: Dr Rita Sharma 
School of Life Sciences, Jawaharlal Nehru University 
Language Editor: Namrata Dhaka 
Department/College: Department of Genetics, University 
of Delhi  South Campus 
Lesson Editor: Dr Rama Sisodia, Fellow in Botany ILLL 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            2 
 
Learning Outcomes 
The chapter is aimed to enable the readers to learn about the following: 
? Basic facilities required to start a plant tissue culture laboratory.  
? Function of all these facilities and their importance in context of successful tissue 
culture practices. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            3 
 
Chapter: Laboratory Requirements in Plant Tissue Culture 
Table of Contents  
? Introduction 
? Basic facilities required in a tissue culture laboratory 
? Washing area 
? Glassware/Plastic ware 
? Area for media preparation 
? Transfer area 
? Culture room and incubators 
? Greenhouse 
? Basic laboratory equipment and apparatus 
? Summary  
? Exercise/ Practice 
? Glossary 
? References/ Bibliography/ Further Reading 
 
 
 
 
 
 
 
 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            4 
 
Introduction 
Plant tissue culture techniques have played a pivotal role in addressing many questions of 
basic and applied fields of plant science. Initially, this technique was used for studying the 
roles of hormones in cytodifferentiation and organogenesis. It is now also used for the 
functional characterization of genes, elucidation of underlying regulatory mechanisms by 
developing genetically modified (transgenic) plants. Due to their intensive use in various 
fields of applied plant science, plant tissue culture techniques have also revolutionized the 
agriculture sector in modern times. For example, selected plants tissues/cells are cultured 
as suspended cells to produce plant products. Additionally, transgenic plants with enhanced 
agronomic traits such as high carotenoids (golden rice: Ye et al., 2000) or longer shelf life 
(Flavr Savr tomatoes: Redenbough et al., 1992) have been produced. Further, tissue culture 
methods also help in developing the somatic haploid embryos. Such somatic haploid 
embryos can subsequently be used for the production of homozygous plants. Hence, tissue 
culture techniques have played an important role in the progression of academic 
and applied plant science. 
The major applications of plant tissue culture techniques are: 
? The large scale production of ornamental plants. 
? Conservation of endangered plant species. 
? Production of plant-derived secondary metabolites and recombinant proteins on a large-
scale in liquid culture of plant cells in bioreactors. 
? Production of novel hybrids by either fusing protoplasts of distantly-related species or 
embryo-rescue technique. 
? Molecular, pharmacological and biochemical investigations of different aspects of plant 
growth and development such as in vitro flowering. 
? For the induction of polyploidy, by treating plants with of antimitotic agents such 
as colchicine. 
? To produce virus free plants from virus-infected stock, such as potatoes by culturing 
meristem/tip culture. 
? Production of genetically modified crops with improved agronomic traits. 
 
 
Laboratory Requirements in Plant Tissue Culture 
 
                                Institute of Lifelong Learning, University of Delhi                            5 
 
Brief overview of the techniques involved in Plant tissue culture 
 
 
Figure: Steps involved in tissue culture. A number of tissues can be used as explants 
and used for in vitro culture to regenerate whole plants by various methods of tissue 
culture. 
Source: ILLL Inhouse 
 
Figure: A general scheme of plant tissue culture. The figure shows the different steps 
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FAQs on Laboratory requirements of plant tissue culture - Plant Biotechnology - Botany

1. What are the basic requirements for a plant tissue culture laboratory?
Ans. A plant tissue culture laboratory requires the following basic requirements: - Sterile work environment: A sterile work area is crucial to prevent contamination of the plant tissue cultures. This includes laminar flow hoods or clean benches, autoclaves for sterilization, and proper handling techniques. - Growth media and chemicals: Specialized growth media containing essential nutrients and growth regulators are necessary for the successful growth of plant tissues. Chemicals like agar, sucrose, and vitamins are also required. - Equipment: The laboratory needs various equipment such as microscopes, incubators, pH meters, and balances for weighing chemicals. - Lighting and temperature control: Adequate lighting and temperature control are essential for optimal growth of plant tissues. Plant tissue cultures often require specific temperature and light conditions for growth. - Proper disposal system: Proper disposal of waste materials, including used culture media and contaminated materials, is necessary to maintain a clean and safe laboratory environment.
2. How can contamination be prevented in a plant tissue culture laboratory?
Ans. Contamination in a plant tissue culture laboratory can be prevented by following these measures: - Sterilization: All equipment, growth media, and containers used in the laboratory should be sterilized. Autoclaving is the most common method for sterilization. - Clean work area: The work area should be regularly cleaned and disinfected. Wiping down surfaces with disinfectants like ethanol or bleach can help prevent contamination. - Proper handling techniques: Careful handling of plant tissues, tools, and solutions is important to minimize the risk of introducing contaminants. Techniques such as flaming or dipping tools in alcohol before use can help reduce contamination. - Controlled access: Limiting access to the laboratory and ensuring that only trained personnel are allowed can minimize the risk of contamination from outside sources. - Regular monitoring: Regularly checking cultures for signs of contamination, such as discoloration or mold growth, allows for early detection and immediate action to prevent further contamination.
3. What is the role of growth regulators in plant tissue culture?
Ans. Growth regulators play a crucial role in plant tissue culture. They are chemical compounds that regulate plant growth and development. In tissue culture, growth regulators are used to manipulate the growth of plant tissues. They can be classified into two main types: auxins and cytokinins. Auxins promote cell division and differentiation. They are used to initiate the formation of roots and shoots from explants, induce callus formation, and stimulate the growth of embryos. Examples of auxins commonly used in tissue culture are indole-3-acetic acid (IAA) and 1-naphthaleneacetic acid (NAA). Cytokinins, on the other hand, promote cell division and shoot formation. They are used to control the ratio of shoots to roots and enhance the multiplication of plant tissues. Commonly used cytokinins in tissue culture include kinetin, zeatin, and benzyladenine. By manipulating the concentrations and combinations of growth regulators, tissue culture researchers can control the growth and development of plant tissues, leading to the production of new plants or the multiplication of desired traits.
4. Why is a sterile work environment important in plant tissue culture?
Ans. A sterile work environment is crucial in plant tissue culture for several reasons: - Preventing contamination: Contamination of plant tissue cultures with microorganisms, such as bacteria and fungi, can hinder the growth and development of the tissues. A sterile environment helps prevent the introduction of contaminants that could compromise the integrity of the cultures. - Maintaining genetic stability: Plant tissue cultures are often used for clonal propagation, where identical copies of a plant are produced. Any contamination can introduce genetic variation, leading to inconsistent results or loss of desired traits. - Ensuring reproducibility: In research and commercial applications of tissue culture, reproducibility is essential. A sterile environment reduces the variables that could affect the outcomes of experiments or production processes, leading to more consistent and reliable results. - Protecting the laboratory staff: Some microorganisms present in contaminated cultures can be harmful to laboratory personnel. A sterile work environment minimizes the risk of exposure to potentially harmful pathogens. - Saving time and resources: Contaminated cultures often need to be discarded, leading to wastage of time, effort, and resources invested in their preparation. A sterile work environment helps avoid such losses and ensures the efficient use of resources.
5. How can lighting and temperature be controlled in a plant tissue culture laboratory?
Ans. Lighting and temperature control in a plant tissue culture laboratory can be achieved through various methods: - Lighting control: Tissue culture rooms often use artificial light sources, such as fluorescent or LED lights, to provide the necessary light for plant growth. The lighting duration and intensity can be controlled using timers or dimmers to mimic natural day-night cycles or optimize growth conditions for specific plant species. - Temperature control: Climate-controlled incubators or growth chambers are used to maintain the desired temperature for plant tissue cultures. These chambers are equipped with heating and cooling systems, as well as temperature controllers, to maintain a stable and precise temperature range. The temperature can be adjusted based on the specific requirements of the plant species being cultured. - Sensors and data loggers: Sensors can be installed in tissue culture rooms or chambers to monitor and maintain optimal lighting and temperature conditions. Data loggers can record temperature and lighting levels over time, allowing for analysis and adjustment if needed. - Thermoregulation techniques: In addition to incubators and growth chambers, other thermoregulation techniques can be employed, such as using cooling fans or heat pads, to fine-tune temperature control in specific areas or for specific cultures within the laboratory. These techniques help ensure that the growth environment remains within the desired temperature range for successful tissue culture.
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