Laboratory requirements of plant tissue culture | Plant Biotechnology - Botany PDF Download

 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