Page 1
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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