I hereby declare that a dissertation work entitled ―Micropropagation studies on Bambusa tulda plant through nodal explant” Submitted to university in fulfillment for the award of degree in Bachelors Of Science (forestry) is carried out by me at State Research Institute Jabalpur Madhya Pradesh.
Tissue culture involves growing plant and animal cells, tissues, or organs artificially in a sterile environment with a nutrient medium. It allows for cloning and mass propagation of plants and animals. Tissue culture requires selecting appropriate explant tissue, developing a sterile culture medium with growth regulators, and regularly subculturing and multiplying the cells. It has many applications like rapidly propagating desirable crops and endangered species, providing pathogen-free plants, selecting for improved varieties, and studying cell processes. Animal tissue culture differs in requiring serum, having limited cell divisions, and posing some biohazard risks. Its uses include growing viruses, producing monoclonal antibodies, and facilitating genetic studies.
Conservation and preservation of germplasmIñnøcènt ÅñDi
The document discusses germplasm conservation, including both ex situ and in situ methods. Ex situ conservation involves maintaining genetic resources outside their natural habitat, such as in seed banks, field gene banks, DNA banks, botanical gardens, and through in vitro and cryopreservation methods. In situ conservation preserves species in their natural environments through biosphere reserves, national parks, wildlife sanctuaries, and on-farm conservation. Cryopreservation is described as a method to bring plant cells and tissues to a zero metabolism state through freezing at very low temperatures in liquid nitrogen.
- Ovule culture involves aseptically isolating ovules from ovaries and growing them on defined nutrient media under controlled conditions. This allows for studying embryo development from zygote to mature embryo.
- Ovaries and unfertilized ovules can be sources for haploid or doubled haploid production. An ovule contains a megaspore or egg cell that can be fertilized to form a zygote and eventually a mature embryo.
- The protocol involves collecting unfertilized or fertilized ovules, surface sterilizing the ovaries, isolating ovules using a spatula, and incubating them on solid or liquid media in light or dark conditions.
OVARY CULTURE:-
"the in-vitro culturing of ovaries in an aseptic condition from the pollinated or un-pollinated flowers, in an appropriate nutrient medium and under optimal conditions." And
OVULE CULTURE:-
"Ovule culture is an experimental system by which ovules are aseptically isolated from the ovary and are grown aseptically on chemically defined nutrient medium under controlled conditions."
Acclimatization or acclimatisation (also called acclimation or acclimatation) is the process in which an individual organism adjusts to a change in its environment (such as a change in altitude, temperature, humidity, photoperiod, or pH), allowing it to maintain performance across a range of environmental conditions
Invitro mutation selection for biotic stresses in Plantsamvannan
In-vitro selection is a somaclonal variation method that uses a selection agent or particular condition to select for somaclones with a desired character. Various mutagens like gamma irradiation, chemicals, and transposons can be used to induce mutations in vitro. Somatic embryogenesis is advantageous for in-vitro selection as it allows treatment of large populations and rapid generation of non-chimeric plants. Chemical mutagens are commonly used for in-vitro selection due to ease of handling. In-vitro selection has been used successfully to obtain disease resistance in various crop species like tobacco, rice, wheat, and potato.
1. Callus culture involves growing undifferentiated plant cells and tissues on a nutrient medium under sterile conditions. This allows for the production of genetically identical clones without seeds or pollination.
2. A callus is an unorganized mass of cells formed from injured or cultured plant tissue. Successful callus culture requires selecting an explant, preparing sterile culture media, and regulating hormone levels to induce cell proliferation.
3. Callus cultures are maintained through periodic sub-culturing to replenish nutrients and prevent toxicity. The growth and characteristics of callus tissue can provide insights into plant cell metabolism, differentiation, and pathways for genetic engineering applications.
Tissue culture involves growing plant and animal cells, tissues, or organs artificially in a sterile environment with a nutrient medium. It allows for cloning and mass propagation of plants and animals. Tissue culture requires selecting appropriate explant tissue, developing a sterile culture medium with growth regulators, and regularly subculturing and multiplying the cells. It has many applications like rapidly propagating desirable crops and endangered species, providing pathogen-free plants, selecting for improved varieties, and studying cell processes. Animal tissue culture differs in requiring serum, having limited cell divisions, and posing some biohazard risks. Its uses include growing viruses, producing monoclonal antibodies, and facilitating genetic studies.
Conservation and preservation of germplasmIñnøcènt ÅñDi
The document discusses germplasm conservation, including both ex situ and in situ methods. Ex situ conservation involves maintaining genetic resources outside their natural habitat, such as in seed banks, field gene banks, DNA banks, botanical gardens, and through in vitro and cryopreservation methods. In situ conservation preserves species in their natural environments through biosphere reserves, national parks, wildlife sanctuaries, and on-farm conservation. Cryopreservation is described as a method to bring plant cells and tissues to a zero metabolism state through freezing at very low temperatures in liquid nitrogen.
- Ovule culture involves aseptically isolating ovules from ovaries and growing them on defined nutrient media under controlled conditions. This allows for studying embryo development from zygote to mature embryo.
- Ovaries and unfertilized ovules can be sources for haploid or doubled haploid production. An ovule contains a megaspore or egg cell that can be fertilized to form a zygote and eventually a mature embryo.
- The protocol involves collecting unfertilized or fertilized ovules, surface sterilizing the ovaries, isolating ovules using a spatula, and incubating them on solid or liquid media in light or dark conditions.
OVARY CULTURE:-
"the in-vitro culturing of ovaries in an aseptic condition from the pollinated or un-pollinated flowers, in an appropriate nutrient medium and under optimal conditions." And
OVULE CULTURE:-
"Ovule culture is an experimental system by which ovules are aseptically isolated from the ovary and are grown aseptically on chemically defined nutrient medium under controlled conditions."
Acclimatization or acclimatisation (also called acclimation or acclimatation) is the process in which an individual organism adjusts to a change in its environment (such as a change in altitude, temperature, humidity, photoperiod, or pH), allowing it to maintain performance across a range of environmental conditions
Invitro mutation selection for biotic stresses in Plantsamvannan
In-vitro selection is a somaclonal variation method that uses a selection agent or particular condition to select for somaclones with a desired character. Various mutagens like gamma irradiation, chemicals, and transposons can be used to induce mutations in vitro. Somatic embryogenesis is advantageous for in-vitro selection as it allows treatment of large populations and rapid generation of non-chimeric plants. Chemical mutagens are commonly used for in-vitro selection due to ease of handling. In-vitro selection has been used successfully to obtain disease resistance in various crop species like tobacco, rice, wheat, and potato.
1. Callus culture involves growing undifferentiated plant cells and tissues on a nutrient medium under sterile conditions. This allows for the production of genetically identical clones without seeds or pollination.
2. A callus is an unorganized mass of cells formed from injured or cultured plant tissue. Successful callus culture requires selecting an explant, preparing sterile culture media, and regulating hormone levels to induce cell proliferation.
3. Callus cultures are maintained through periodic sub-culturing to replenish nutrients and prevent toxicity. The growth and characteristics of callus tissue can provide insights into plant cell metabolism, differentiation, and pathways for genetic engineering applications.
Aseptic techniques in plant tissue culturekumarkanika
Importance of practising Aseptic Techniques in plant tissue culture and what are these techniques what precautions should be taken when doing tissue culture
Marker Assisted Selection in Crop BreedingPawan Chauhan
Marker Assisted Selection is a value addition to conventional methods of Crop Breeding. It has been gaining importance in plant breeding with new generation of plant breeders and to get accurate and fast desired result from plant breeding.
This document discusses plant tissue culture and micropropagation techniques. It begins with an introduction to plant tissue culture and defines it as the growth of plant cells, tissues or organs in a sterile nutrient culture medium under controlled conditions. The document then provides a brief history of plant tissue culture, outlines the process of micropropagation including initiation, multiplication, rooting and acclimatization stages. It discusses using micropropagation to produce disease-free plants by culturing apical meristems and provides examples of various plants that have been made disease-free through this method. The advantages and disadvantages of micropropagation are also summarized along with current applications and future prospects of plant tissue culture techniques.
1) Clonal propagation is the multiplication of genetically identical copies of a plant cultivar through asexual reproduction. This can be done in vivo through methods like cutting, layering, and grafting, or in vitro through tissue culture techniques.
2) Micropropagation through tissue culture involves taking explants like shoot tips or meristems and culturing them on nutrient media to induce multiplication. Common methods are meristem culture, organogenesis, and somatic embryogenesis.
3) Clonal propagation has advantages like producing true-to-type plants, allowing off-season production, reducing life cycles, enabling large-scale production, and facilitating germplasm conservation and genetic transformation. However, it can be costly and labor intensive
In vitro pollination involves pollinating pistils or ovules that have been cultured in a nutrient medium such as Nitsch's medium. This technique can help overcome pre-fertilization barriers to hybridization between plant species. Key steps include sterilizing flower parts, collecting pollen, and applying pollen to excised pistils, ovaries, ovules, or stigmas depending on the method. Factors like culture medium, temperature, genotype, and physiological state of the explant can influence seed set. In vitro pollination has applications in plant breeding like overcoming self-incompatibility or cross-incompatibility barriers and producing haploid plants or hybrids.
This document discusses plant tissue culture, in vitro pollination, and in vitro fertilization. It covers the history of plant tissue culture, types of tissue culture including meristem culture and callus culture, selection of explants, and the basic steps of tissue culture including inoculation and induction of callus. It also describes different types of in vitro pollination and techniques for in vitro fertilization in maize plants, including isolation of sperm and egg cells. Applications of these techniques include micropropagation of ornamental plants and production of artificial seeds.
This document summarizes research on somatic embryogenesis in rice. It describes the process of somatic embryogenesis, including the stages of embryogenesis and factors that affect it. The methodology section outlines the materials and methods used, including collecting rice seeds as explants, sterilizing them, and culturing them on callus induction and embryo germination media with different concentrations of plant growth regulators like 2,4-D, BAP and NAA. The goal is to develop an efficient system for somatic embryogenesis and plant regeneration in rice.
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
Tissue culture techniques can be used for crop improvement in several ways:
1) They allow for wide hybridization through techniques like in vitro fertilization, embryo culture, and protoplast fusion to overcome genetic barriers.
2) They enable the production of haploids which are useful for developing homozygous lines.
3) Somaclonal variation generated in tissue culture can provide traits with agricultural importance.
4) Micropropagation allows for large-scale propagation of disease-free plants.
1. The document discusses various techniques of in vitro plant tissue culture including organ culture, seed culture, meristem culture, embryo culture, ovary culture, anther culture, callus culture, cell suspension culture, and protoplast culture.
2. Key steps in in vitro culture include isolation of explant tissues, regeneration and callus formation, embryogenesis, and organogenesis.
3. Applications of these techniques include virus-free plant production, increasing seed germination efficiency, producing somatic embryos and haploid plants, enabling single cell studies, and facilitating somatic hybridization.
Plant Tissue Culture (Nucleic Acids, Amino Acids, Callus Culture, Transgenic Plants, Embryo Rescue, Embryonic Tissues, Cometabolism, Fungi and Actinomycetes, Grampositive Rods, Cloning Vectors, Biodegradation, Batch Cultures, Organ Culture)
Plants cell tissue culture is a rapidly developing technology which holds promise of restructuring agricultural and forestry practices. During the last two decades cell culture have made considerable advanced in the field of agriculture, horticulture, plant breeding, forestry, somatic cell genetics, phytopathology etc. Plant cells can be grown in isolation from intact plants in tissue culture systems. The cells have the characteristics of callus cells, rather than other plant cell types. These are the cells that appear on cut surfaces when a plant is wounded and which gradually cover and seal the damaged area.
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106-E, Kamla Nagar, Opp. Spark Mall,
New Delhi-110007, India.
Email: npcs.ei@gmail.com , info@entrepreneurindia.co
Tel: +91-11-23843955, 23845654, 23845886, 8800733955
Mobile: +91-9811043595
Website: www.entrepreneurindia.co , www.niir.org
Tags
Plant Tissue Culture in India, Commercialization of Plant Tissue Culture in India, Role of Plant Tissue Culture in Agriculture, Plant Tissue Culture Industry in India, Industrial Plant Tissue Culture, Tissue Culture in Agriculture, Plant Tissue Culture, Tissue Culture, Cell Culture and Tissue Culture, Tissue Culture and Cell Culture, Tissue Culture in Plants, Plant Cell and Tissue Culture, Commercial Plant Tissue Culture in India, Plant Tissue Culture Business Plan, Plant Tissue Culture and Biotechnology, Tissue Culture Plants, Plant Tissue Culture Business Plan, Business Opportunities in Plant Tissue Culture, Tissue Culture Methods, Cybrid Production, Process of Cybrids Production, Production of Cybrids, Production of Cybrid Plants, Production of Haploid Plants, Haploid Production, Plant Secondary Metabolism, Production of Secondary Metabolites, Production of Secondary Metabolites Using Plant Cell Cultures, Plant Tissue Cultures in Production of Secondary Metabolites, Secondary Metabolites Production, Production of Somatic Hybrid Plants, Somatic Hybridization of Plants, Somatic Hybrid, Somatic Hybrid Production, Production of Enriched Biomass, Enrichment on Biomass Production, Formulation of Tissue Culture Medium, Collection of Explant Materials, Subculture of Callus, Regeneration of Plants from Callus, Preparation of Chick Embryo Extract, Preparation of Embryo Extract from Young Embryos, Preparation of Bovine Embryo Extract, Preparation of Eagles Medium, Media for Plant Tissues, Organ Culture, Preparation of Trypsinised Embryonic Carcass, Enrichment Culture Methods, Genetic Modification of Industrial Microorganisms Mutation
Micropropagation
steps of micropropagation
system used to regenerate plantlets by micropropagation
methods of micropropagation
embrogensis
organogenesis
bud culture
How does micropropogation work?
Examples with flow diagrams
Advantages & Disadvantages
Applications
Reference
Somatic hybridisation involves the fusion of somatic protoplasts from two different plant species or varieties to create a hybrid plant. It allows for novel combinations of genes from unrelated species and can overcome barriers to sexual crossing. Protoplasts are isolated from plant tissues enzymatically or mechanically and then fused using techniques like PEG or electrofusion. Hybrid cells are selected and cultured, and plants are regenerated from hybrid calli. Somatic hybridisation has applications in genetic recombination of asexual plants, transferring desirable traits between species, and studying organelle genomes.
This document outlines the procedure for tissue culture and micropropagation through in vitro culture. It involves 3 major stages: 1) preparation which includes explant selection, sterilization and inoculation onto nutrient medium. 2) Sub-culturing to induce callus formation. 3) Organogenesis where the callus is transferred to auxin-rich medium to induce root formation, followed by hardening in a greenhouse and transplantation to soil. Key steps include explant sterilization using antiseptic agents, preparation of solidified nutrient medium, incubation for callus induction, sub-culturing every 28 days to prevent nutrient depletion, and a final acclimatization and transplantation stage.
Embryo culture is a laboratory method for producing plant lets from a fertilized or unfertilized embryo in invitro condition. there are several advantages are associated with the embryo culture like production of haploid plants, making distant crosses successful, sometimes aborted embryos can be rescued from a unsuccessful hybridization.
Somaclonal and gametoclonal variation refer to genetic variations that arise in plants regenerated from cell and tissue cultures. There are two main types - somaclonal variation originating from somatic cells, and gametoclonal variation from gametic cells like pollen. Variations can be induced through long term culture, exposure to mutagens, or selection in media containing inhibitors or toxins. Somaclonal variants are isolated and screened using cytological, biochemical, and molecular markers to identify desirable heritable traits for commercial use in plant breeding programs.
This practical aims to induce callus formation from different plant explants. Students will take explants like leaf, stem, and root segments from selected horticultural crops and culture them on nutrient media supplemented with plant growth regulators, like auxins and cytokinins, to induce callus tissue. The success of callus induction will depend on the explant type and plant species as well as the plant growth regulator concentrations used in the culture medium.
This document provides information about a horticulture course focusing on biotechnology. The course code is HRT 552 and is titled "BIOTECHNOLOGY OF HORTICULTURAL CROPS." Practical 9 of the course covers mass multiplication through direct organogenesis.
Somatic embryogenesis is the process where embryos form from somatic (non-reproductive) plant cells in vitro. It is an important biotechnological tool that allows for clonal propagation, genetic transformation, and other applications. The first observation of somatic embryogenesis was in carrot cells in 1958. Somatic embryogenesis occurs through direct or indirect pathways and involves induction, development, and maturation stages. It has advantages over zygotic embryogenesis like a higher propagation rate and applications in synthetic seed production and genetic engineering.
This document discusses embryo culture and embryo rescue techniques. It begins by defining an embryo and explaining that embryo culture involves growing plant embryos in artificial media to enhance survival. Embryo rescue involves culturing immature embryos to prevent abortion, especially for interspecific or intergeneric hybrids where endosperm development fails. The key steps of embryo culture include excising embryos, placing them in sterile media with suitable temperature, light and nutrients, and transferring viable plantlets to soil. Embryo culture has applications in shortening breeding cycles, overcoming dormancy, producing sterile seeds, and rescuing distant hybrids.
Plant tissue culture is the process of growing plant cells, tissues or organs in sterile conditions on a nutrient medium. It has many applications like germplasm preservation of endangered plants, producing disease-free plants through micropropagation, and creating novel hybrids through protoplast fusion and somatic hybridization. However, issues remain like genetic instability of hybrids and lack of efficient selection methods. Overall, tissue culture is a valuable biotechnology tool with potential for crop improvement and conservation efforts.
Plant tissue culture is the process of growing plant cells, tissues or organs in sterile conditions on a nutrient medium. It has many applications like germplasm preservation of endangered plants, genetic improvement of crops, and production of secondary metabolites. The basic steps include selection of explant, initiation of culture on growth media, multiplication through cell division, and rooting and transfer to soil. Technologies such as micropropagation, somatic hybridization, and cryopreservation have been commercialized for mass propagation of crops. However, issues remain regarding genetic stability, selection of fused products, and regeneration efficiency.
Aseptic techniques in plant tissue culturekumarkanika
Importance of practising Aseptic Techniques in plant tissue culture and what are these techniques what precautions should be taken when doing tissue culture
Marker Assisted Selection in Crop BreedingPawan Chauhan
Marker Assisted Selection is a value addition to conventional methods of Crop Breeding. It has been gaining importance in plant breeding with new generation of plant breeders and to get accurate and fast desired result from plant breeding.
This document discusses plant tissue culture and micropropagation techniques. It begins with an introduction to plant tissue culture and defines it as the growth of plant cells, tissues or organs in a sterile nutrient culture medium under controlled conditions. The document then provides a brief history of plant tissue culture, outlines the process of micropropagation including initiation, multiplication, rooting and acclimatization stages. It discusses using micropropagation to produce disease-free plants by culturing apical meristems and provides examples of various plants that have been made disease-free through this method. The advantages and disadvantages of micropropagation are also summarized along with current applications and future prospects of plant tissue culture techniques.
1) Clonal propagation is the multiplication of genetically identical copies of a plant cultivar through asexual reproduction. This can be done in vivo through methods like cutting, layering, and grafting, or in vitro through tissue culture techniques.
2) Micropropagation through tissue culture involves taking explants like shoot tips or meristems and culturing them on nutrient media to induce multiplication. Common methods are meristem culture, organogenesis, and somatic embryogenesis.
3) Clonal propagation has advantages like producing true-to-type plants, allowing off-season production, reducing life cycles, enabling large-scale production, and facilitating germplasm conservation and genetic transformation. However, it can be costly and labor intensive
In vitro pollination involves pollinating pistils or ovules that have been cultured in a nutrient medium such as Nitsch's medium. This technique can help overcome pre-fertilization barriers to hybridization between plant species. Key steps include sterilizing flower parts, collecting pollen, and applying pollen to excised pistils, ovaries, ovules, or stigmas depending on the method. Factors like culture medium, temperature, genotype, and physiological state of the explant can influence seed set. In vitro pollination has applications in plant breeding like overcoming self-incompatibility or cross-incompatibility barriers and producing haploid plants or hybrids.
This document discusses plant tissue culture, in vitro pollination, and in vitro fertilization. It covers the history of plant tissue culture, types of tissue culture including meristem culture and callus culture, selection of explants, and the basic steps of tissue culture including inoculation and induction of callus. It also describes different types of in vitro pollination and techniques for in vitro fertilization in maize plants, including isolation of sperm and egg cells. Applications of these techniques include micropropagation of ornamental plants and production of artificial seeds.
This document summarizes research on somatic embryogenesis in rice. It describes the process of somatic embryogenesis, including the stages of embryogenesis and factors that affect it. The methodology section outlines the materials and methods used, including collecting rice seeds as explants, sterilizing them, and culturing them on callus induction and embryo germination media with different concentrations of plant growth regulators like 2,4-D, BAP and NAA. The goal is to develop an efficient system for somatic embryogenesis and plant regeneration in rice.
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
Tissue culture techniques can be used for crop improvement in several ways:
1) They allow for wide hybridization through techniques like in vitro fertilization, embryo culture, and protoplast fusion to overcome genetic barriers.
2) They enable the production of haploids which are useful for developing homozygous lines.
3) Somaclonal variation generated in tissue culture can provide traits with agricultural importance.
4) Micropropagation allows for large-scale propagation of disease-free plants.
1. The document discusses various techniques of in vitro plant tissue culture including organ culture, seed culture, meristem culture, embryo culture, ovary culture, anther culture, callus culture, cell suspension culture, and protoplast culture.
2. Key steps in in vitro culture include isolation of explant tissues, regeneration and callus formation, embryogenesis, and organogenesis.
3. Applications of these techniques include virus-free plant production, increasing seed germination efficiency, producing somatic embryos and haploid plants, enabling single cell studies, and facilitating somatic hybridization.
Plant Tissue Culture (Nucleic Acids, Amino Acids, Callus Culture, Transgenic Plants, Embryo Rescue, Embryonic Tissues, Cometabolism, Fungi and Actinomycetes, Grampositive Rods, Cloning Vectors, Biodegradation, Batch Cultures, Organ Culture)
Plants cell tissue culture is a rapidly developing technology which holds promise of restructuring agricultural and forestry practices. During the last two decades cell culture have made considerable advanced in the field of agriculture, horticulture, plant breeding, forestry, somatic cell genetics, phytopathology etc. Plant cells can be grown in isolation from intact plants in tissue culture systems. The cells have the characteristics of callus cells, rather than other plant cell types. These are the cells that appear on cut surfaces when a plant is wounded and which gradually cover and seal the damaged area.
See more
https://goo.gl/pXccQD
https://goo.gl/MNnSqw
https://goo.gl/QgJiqW
Contact us:
Niir Project Consultancy Services
106-E, Kamla Nagar, Opp. Spark Mall,
New Delhi-110007, India.
Email: npcs.ei@gmail.com , info@entrepreneurindia.co
Tel: +91-11-23843955, 23845654, 23845886, 8800733955
Mobile: +91-9811043595
Website: www.entrepreneurindia.co , www.niir.org
Tags
Plant Tissue Culture in India, Commercialization of Plant Tissue Culture in India, Role of Plant Tissue Culture in Agriculture, Plant Tissue Culture Industry in India, Industrial Plant Tissue Culture, Tissue Culture in Agriculture, Plant Tissue Culture, Tissue Culture, Cell Culture and Tissue Culture, Tissue Culture and Cell Culture, Tissue Culture in Plants, Plant Cell and Tissue Culture, Commercial Plant Tissue Culture in India, Plant Tissue Culture Business Plan, Plant Tissue Culture and Biotechnology, Tissue Culture Plants, Plant Tissue Culture Business Plan, Business Opportunities in Plant Tissue Culture, Tissue Culture Methods, Cybrid Production, Process of Cybrids Production, Production of Cybrids, Production of Cybrid Plants, Production of Haploid Plants, Haploid Production, Plant Secondary Metabolism, Production of Secondary Metabolites, Production of Secondary Metabolites Using Plant Cell Cultures, Plant Tissue Cultures in Production of Secondary Metabolites, Secondary Metabolites Production, Production of Somatic Hybrid Plants, Somatic Hybridization of Plants, Somatic Hybrid, Somatic Hybrid Production, Production of Enriched Biomass, Enrichment on Biomass Production, Formulation of Tissue Culture Medium, Collection of Explant Materials, Subculture of Callus, Regeneration of Plants from Callus, Preparation of Chick Embryo Extract, Preparation of Embryo Extract from Young Embryos, Preparation of Bovine Embryo Extract, Preparation of Eagles Medium, Media for Plant Tissues, Organ Culture, Preparation of Trypsinised Embryonic Carcass, Enrichment Culture Methods, Genetic Modification of Industrial Microorganisms Mutation
Micropropagation
steps of micropropagation
system used to regenerate plantlets by micropropagation
methods of micropropagation
embrogensis
organogenesis
bud culture
How does micropropogation work?
Examples with flow diagrams
Advantages & Disadvantages
Applications
Reference
Somatic hybridisation involves the fusion of somatic protoplasts from two different plant species or varieties to create a hybrid plant. It allows for novel combinations of genes from unrelated species and can overcome barriers to sexual crossing. Protoplasts are isolated from plant tissues enzymatically or mechanically and then fused using techniques like PEG or electrofusion. Hybrid cells are selected and cultured, and plants are regenerated from hybrid calli. Somatic hybridisation has applications in genetic recombination of asexual plants, transferring desirable traits between species, and studying organelle genomes.
This document outlines the procedure for tissue culture and micropropagation through in vitro culture. It involves 3 major stages: 1) preparation which includes explant selection, sterilization and inoculation onto nutrient medium. 2) Sub-culturing to induce callus formation. 3) Organogenesis where the callus is transferred to auxin-rich medium to induce root formation, followed by hardening in a greenhouse and transplantation to soil. Key steps include explant sterilization using antiseptic agents, preparation of solidified nutrient medium, incubation for callus induction, sub-culturing every 28 days to prevent nutrient depletion, and a final acclimatization and transplantation stage.
Embryo culture is a laboratory method for producing plant lets from a fertilized or unfertilized embryo in invitro condition. there are several advantages are associated with the embryo culture like production of haploid plants, making distant crosses successful, sometimes aborted embryos can be rescued from a unsuccessful hybridization.
Somaclonal and gametoclonal variation refer to genetic variations that arise in plants regenerated from cell and tissue cultures. There are two main types - somaclonal variation originating from somatic cells, and gametoclonal variation from gametic cells like pollen. Variations can be induced through long term culture, exposure to mutagens, or selection in media containing inhibitors or toxins. Somaclonal variants are isolated and screened using cytological, biochemical, and molecular markers to identify desirable heritable traits for commercial use in plant breeding programs.
This practical aims to induce callus formation from different plant explants. Students will take explants like leaf, stem, and root segments from selected horticultural crops and culture them on nutrient media supplemented with plant growth regulators, like auxins and cytokinins, to induce callus tissue. The success of callus induction will depend on the explant type and plant species as well as the plant growth regulator concentrations used in the culture medium.
This document provides information about a horticulture course focusing on biotechnology. The course code is HRT 552 and is titled "BIOTECHNOLOGY OF HORTICULTURAL CROPS." Practical 9 of the course covers mass multiplication through direct organogenesis.
Somatic embryogenesis is the process where embryos form from somatic (non-reproductive) plant cells in vitro. It is an important biotechnological tool that allows for clonal propagation, genetic transformation, and other applications. The first observation of somatic embryogenesis was in carrot cells in 1958. Somatic embryogenesis occurs through direct or indirect pathways and involves induction, development, and maturation stages. It has advantages over zygotic embryogenesis like a higher propagation rate and applications in synthetic seed production and genetic engineering.
This document discusses embryo culture and embryo rescue techniques. It begins by defining an embryo and explaining that embryo culture involves growing plant embryos in artificial media to enhance survival. Embryo rescue involves culturing immature embryos to prevent abortion, especially for interspecific or intergeneric hybrids where endosperm development fails. The key steps of embryo culture include excising embryos, placing them in sterile media with suitable temperature, light and nutrients, and transferring viable plantlets to soil. Embryo culture has applications in shortening breeding cycles, overcoming dormancy, producing sterile seeds, and rescuing distant hybrids.
Plant tissue culture is the process of growing plant cells, tissues or organs in sterile conditions on a nutrient medium. It has many applications like germplasm preservation of endangered plants, producing disease-free plants through micropropagation, and creating novel hybrids through protoplast fusion and somatic hybridization. However, issues remain like genetic instability of hybrids and lack of efficient selection methods. Overall, tissue culture is a valuable biotechnology tool with potential for crop improvement and conservation efforts.
Plant tissue culture is the process of growing plant cells, tissues or organs in sterile conditions on a nutrient medium. It has many applications like germplasm preservation of endangered plants, genetic improvement of crops, and production of secondary metabolites. The basic steps include selection of explant, initiation of culture on growth media, multiplication through cell division, and rooting and transfer to soil. Technologies such as micropropagation, somatic hybridization, and cryopreservation have been commercialized for mass propagation of crops. However, issues remain regarding genetic stability, selection of fused products, and regeneration efficiency.
This document summarizes plant tissue culture techniques. It discusses that plant tissue culture involves cultivating plant organs, tissues or cells in test tubes with artificial media. The basic requirements for plant tissue culture include aseptic conditions, temperature control, sub-culturing, and a proper culture medium. The techniques are grouped into embryo culture, meristem culture, anther or pollen culture, tissue and cell culture, and callus culture. Each technique has specific applications such as micropropagation, recovering virus-free stocks, and facilitating germplasm exchange and conservation. Crops that can be produced using these techniques include tomato, brinjal, potato, banana, orchids, and Chinese cabbage.
Tissue culture is a technique where small pieces of plant or animal tissue are cultured in a sterile medium outside of the organism. It was first developed in 1885 and has since been used extensively in medicine, agriculture, and research. It allows for the rapid duplication of plant materials while eliminating diseases and maintaining genetic traits. However, it requires specialized facilities and equipment and reduces genetic diversity.
Plant tissue culture is a method of growing isolated plant cells, tissues or organs in sterile conditions on an artificial nutrient medium. Early attempts were unsuccessful due to lack of knowledge about plant nutrition and hormones. Major advances included Philip White's 1939 report of continuous carrot and tobacco culture in vitro and Folke Skoog's discovery of auxin properties. Today, tissue culture is used commercially to propagate plants like flowers rapidly and introduce new varieties. Successful tissue culture requires sterile conditions, nutrient media tailored to the plant species, and controlling contamination.
This document provides an introduction to plant tissue culture and propagation prepared for third year biology students. It defines key terms related to plant tissue culture such as explant, callus, and nutrient medium. The document discusses the history and development of plant tissue culture techniques dating back to early experiments in the 1900s. It also covers the application of plant tissue culture for cloning plants, eliminating diseases, and producing metabolites, as well as factors that affect culture efficiency such as growth media, environmental conditions, and the explant source.
Plant tissue culture provides several benefits for studying plant growth and development. It allows scientists to isolate plant parts and culture them in vitro, simplifying the study of controlling influences. Some key applications of plant tissue culture include clonal propagation of disease-free plants, studying plant cells' ability to regenerate whole plants from cultured cells, producing genetic variability through somaclonal variation, regenerating plants from pollen to create haploids, and rescuing hybrid embryos. Plant tissue culture also enables fundamental biological research, production of high-value biochemicals, and generation of transgenic plants.
Application of plant tissue culture/ micro-propagationSushil Nyaupane
Tissue culture is the process of growing cells or tissues in sterile conditions. It allows for rapid cloning of plant materials. Plant tissue culture involves excising plant parts and growing them on nutrient media. This allows for mass multiplication of plant materials irrespective of season. Some key developments include Haberlandt's proposal of plant cell culture in 1902, and Murashige and Skoog's nutrient medium in 1962. Micropropagation is now used for conservation of rare species, producing disease-free plants, mutation breeding, and more. The future of this technique remains promising.
Plant tissue culture involves growing plant cells, tissues or organs in an artificial nutrient medium under sterile conditions. Some key points:
1. It allows for the rapid mass propagation of plants through micropropagation and the production of genetically uniform plants.
2. It facilitates the production of disease-free plants through culture of meristems and shoot tips.
3. It enables genetic modification of plants through techniques like protoplast fusion, anther culture and recombinant DNA technology.
The document discusses the history and techniques of plant tissue culture. It begins with early experiments in the 1830s culturing plant cells outside of their natural environment. Significant developments include the discovery of growth hormones auxin and cytokinin in the 1950s, and the development of the Murashige and Skoog medium in 1962. The document outlines the basic requirements and procedures for plant tissue culture, including selecting an explant, preparing sterile media, inoculation, incubation, sub-culturing, and transferring plantlets. Applications include rapid clonal propagation, inducing mutations, producing disease-free plants, and conserving rare species.
Plant tissue culture has been widely employed in area of agriculture, horticulture, forestry and plant breeding. It is an applied biotechnology used for mass propagation, virus elimination, secondary metabolite production and in vitro cloning of plants. Recently, plant tissue culture has been used for the conservation of endangered plant species through short and medium term conservation also known as slow growth and cryopreservation also known as long term conservation. These methods had been effectively used to conserve plant species with recalcitrant seeds or dormant seeds and showed greater advantage over the conventional methods of conservation. At present plant cell culture has made great advances. Possibly the most significant role that plant cell culture has to play in the future will be in its association with transgenic plants. The ability to accelerate the conventional multiplication rate can be of great benefit to many crops countries where a disease or some climatic disaster wipes out crops. Mr. Rohan R. Vakhariya | Rutuja R. Shah "Over Review on Plant Tissue Culture" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-4 | Issue-1 , December 2019, URL: https://www.ijtsrd.com/papers/ijtsrd29619.pdfPaper URL: https://www.ijtsrd.com/pharmacy/other/29619/over-review-on-plant-tissue-culture/mr-rohan-r-vakhariya
The document discusses totipotency in plant cells. Totipotency refers to the ability of single plant cells to regenerate into a whole plant through cell differentiation and tissue culture techniques. The document outlines various tissue culture systems used to study totipotency, including callus culture, suspension culture, single cell culture, and protoplast culture. Factors that influence a cell's ability to express totipotency, such as the explant source and culture conditions, are also discussed.
This document provides an overview of plant tissue culture techniques. It discusses that plant tissue culture involves growing plant cells, tissues or organs in a sterile environment with nutrient media. The techniques rely on two principles - totipotency, the ability of plant cells to regenerate into a whole plant, and plasticity, the ability of plants to alter their growth in response to their environment. Explants from various plant tissues are sterilized and placed on culture media, which are composed of inorganic salts, organic nutrients and plant hormones. The culture media, explant source, and plant species can affect regeneration efficiency. Applications of plant tissue culture include commercial plant production, conservation of rare species, screening for desirable traits like herbicide resistance, and
This document provides an overview of plant tissue culture techniques. It discusses that plant tissue culture involves growing plant cells, tissues or organs in a sterile environment with nutrient media. The fundamental principles are totipotency, the ability of plant cells to regenerate into a whole plant, and plasticity, the ability of plants to alter their growth in response to their environment. Explants are obtained from plant tissues and sterilized before being placed on culture media, which contains inorganic salts, organic nutrients and plant hormones. Common media include White's, Murashige and Skoog (MS), and Gamborg B5. Factors like plant species, genotype, explant source and health influence culture efficiency. Applications include commercial plant production
I have discussed Applications of Plant Tissue Culture under the following subheadings,
1. Micro Propagation
2. Clonal Propagation
3. Production of Genetically Variable Plants
4. Production of Virus Free Plants
5. Plant Breeding
6. Production of Useful Biochemicals
7. Preservation of Plant Genetic Resources
8. Importance of Tissue Culture in Biotechnology
Plant tissue culture is a technique for growing plant cells, tissues or organs in a sterile nutrient medium. It relies on the principles of totipotency, where plant cells can regenerate into a whole plant, and plasticity, the ability of plants to alter their growth. Explants from various plant tissues are sterilized and placed on nutrient media like MS or B5 medium. Factors like genotype, explant source and media composition affect regeneration efficiency. Applications include commercial plant production, conserving endangered species, screening for traits and producing secondary metabolites.
The document discusses tissue culture techniques and their applications in plant breeding. It provides a historical background of tissue culture dating back to the early 20th century. The major steps involved in tissue culture are explained, including selection of explant tissue, sterilization, establishment in culture medium, multiplication through callus formation, root formation, and hardening of plantlets. Key techniques like protoplast culture, haploid culture, and micropropagation are also summarized along with their achievements in generating somatic hybrids, true breeding lines, and large-scale clonal propagation respectively.
To achieve the target of creating a new plant or a plant with desired characteristics, tissue culture is often coupled with recombinant DNA technology. The techniques of plant tissue culture have largely helped in the green revolution by improving the crop yield and quality.
The knowledge obtained from plant tissue cultures has contributed to our understanding of metabolism, growth, differentiation and morphogenesis of plant cells. Further, developments in tissue culture have helped to produce several pathogen-free plants, besides the synthesis of many biologically important compounds, including pharmaceuticals. Because of the wide range of applications, plant tissue culture attracts the attention of molecular biologists, plant breeders and industrialists.
Similar to Plant Tissue Culture..“Micropropagation Studies On Bambusa tulda” (20)
Jammu and Kashmir Forest Department Ramban Forest DivisionManzoor Wani
This document provides details about a hands-on job training program on various forestry topics conducted by the Jammu and Kashmir Forest Department. It discusses the forest types and areas found in Jammu and Kashmir. It also covers the forest demarcation process and prohibited acts in demarcated forests. Finally, it acknowledges and thanks the various officials from the Forest Department who provided guidance and assistance during the training.
NAME : MANZOOR NABI
COURSE : B.SC (FORESTRY)
(Semester- v)
Year:-2015-16
student at mewar university Rajasthan, India
TOPIC: SHIFTING CULTIVATION
SUB : AGROFORESTRY
CONTACT ME.........
E-mail:- manzoornabi57@gmail.com
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Plant Tissue Culture..“Micropropagation Studies On Bambusa tulda”
1. Plant Tissue Culture
Manzoor Nabi Wani
Only A Small Explant Is Enough To Get Millions Of Plants With Extremely High Multiplication Rate
2016
Every People In
India Depends
Directly On
Forests For
Sustenance.
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“Micropropagation Studies On Bambusa tulda”
Under The Supervision Of
(Sr. Scientist)
State Forest Research Institute Jabalpur Madhya Pradesh
Gangrar, Chittorgarh, 312901(Rajasthan)
Year 2016
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DECLARATION
I hereby declare that a dissertation work entitled ―Micropropagation studies on
Bambusa tulda plant through nodal explant” Submitted to university in fulfillment for
the award of degree in Bachelors Of Science (forestry) is carried out by me at State
Research Institute Jabalpur Madhya Pradesh.
Date:
Manzoor Nabi Wani
B.SC VII SEM
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ACKNOWLEDGEMENT
It is my pleasure to acknowledge the help which I received during the work and through
preparing thesis; this dissertation would not have been possible without guidance and
help of several individuals who contributed their valuable assistance in completion of
this study.
Foremost, I would like to express my sincere gratitude to the Director, SFRI
Dr.G.Krishna Murthy (IFS) for giving me an opportunity to be a part of institute.
I am greatly indebted to my Supervisor Dr.S.K.Tiwari, Scientist and Head, Forest
Genetics Plant Propagation and Biotechnology Division, SFRI, Jabalpur (M.P) for his
abiding interest and invaluable guidance.
I express my profound gratitude to Mr. Amit Panday, Senior Research Officer, Forest
Genetic Plant Propagation and Biotechnology Division, SFRI Jabalpur (M.P) whose
guidance helped me to proceed my work.
I am very thankful to my Dr. Vk.Solanki, Head of Department of Forestry and Mewar
University including S.K.Das and Dr.Ashok Gadiya for their precious guidance.
I shall be failing in my duty, if I do not gratefully acknowledge the valuable time and
inspiration given by, Mr. Maneesh Puri Goswami, Mr. Pankaj Saini and Mr. Vineet
Mhera, SFRI,Jabalpur, Whose tips provided me enthusiasm to work hard.
Finally, I must express my profound gratitude to my Family for providing unfailing
support. Last I would like to thank ALMIGHTY ALLAH for the blessings without which I
might not complete my work.
MANZOOR NABI WANI
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CONTENTS
S.No Topic Page No…
01 Introduction 05
02 Review of Literature 35
03 methodology 38
04 Result and Discussion 46
05 Conclusion 50
06 Reference 61
07 Photo gallery 65
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DESCRIPTION:
Biotechnology is a relatively new science with direct applications to the Agriculture
industry. This article describes some of the pros and cons of Biotechnology.
Introduction:
One of the newest, yet controversial fields in science today is biotechnology.
Biotechnology began in the 1970s after the development of genetic engineering that
allowed scientists to modify the genetic material of living cells. Genetic engineering is
the manipulation of DNA molecules to produce modified plants, animals, or other
organisms. DNA is the part of a cell that controls the genetic information of an animal or
plant. DNA is a double-stranded molecule that is present in every cell of an organism.
The genetic information is contained in individual units or sections of DNA called genes.
The genes that are passed from parent to offspring determine the traits that the
offspring will have. Scientists are now able to isolate the gene or genes for the traits
they want in one animal or plant and move them into another. The movement of a gene
from one organism to another is called recombinant DNA technology.
Gene: A unit of hereditary material located on a chromosome.
Genetic engineering: Movement of genes from one cell to another.
Hormones: Chemicals released by cells that affect cells in other parts of the body.
Only a small amount of hormone is required to alter cell metabolism.
Recombinant DNA technology:
An application of genetic engineering in which genetic information from one organism
is spliced into the chromosome of another organism.
Biotechnology:
Use of cells or components of cells to produce products or processes.
Definition of Plant Biotechnology:
In a broad sense:
Plant biotechnology covers many of the tools and techniques that are commonplace
in agriculture and food production.
In a narrow sense:
Biotechnology considers only the new DNA techniques, molecular biology and
reproductive technological applications, like gene manipulation, gene transfer, DNA
genotyping and cloning.
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DIFFERENT SCIENCE FIELDS CONTRIBUTING INTO THE
ADVANCEMENT OF BIOTECHNOLOGY.
Historical Advancement of Biotechnology:
Biotechnology related activities depend on two parameters: technological
advancement and knowledge of available biota. Technological upgradation goes
parallel with the over-all understanding of physical and chemical phenomenon in
different time periods. Hence, Biotechnology starts as early as human have realized the
importance of organism (animal/plants or microbes) to improve their life-style.
Important milestones of Biotechnology
S.No. Time
Period
Major break-through
1. 7000 BC-100CE
fertility.
fertilizer and insecticide respectively
2. Pre-20th Century
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Applications of Biotechnology:
Biotechnology has influenced human life in many ways by inventions to make his life
more comfortable. Many scientific fields contribute to biotechnology and in return it
gives product for their advancement.
Impact of Biotechnology on different fields & human life.
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What is Plant Tissue Culture?
The aseptic culture of plant protoplasts, cells, tissues or organs under conditions which
lead to cell multiplication or regeneration of organs or whole plants ―
Definition: In plant tissue cultures, sterile plant material is cultured under aseptical
conditions in usually defined sterile culture medium often solidified by agar.
Commonly used terms in tissue culture:
Adventitious: development of organs such as buds, leaves, roots, shoots and
somatic embryos from shoot and root tissues and callus.
Agar: Natural gelling agent made from algae.
Aseptic technique: procedures used to prevent the introduction of microorganisms
such as fungi, bacteria, viruses and phytoplasmas into cell, tissue and organ cultures,
and cross contamination of cultures.
Autoclave: A machine capable of sterilizing by steam under pressure.
Axenic culture: a culture without foreign or undesired life forms but may include the
deliberate co-culture with different types of cells, tissues or organisms.
Callus: an unorganized mass of differentiated plant cells.
Cell culture: culture of cells or their maintenance in vitro including the culture of single
cells.
Chemically defined medium: a nutritive solution or substrate for culturing cells in which
each component is specified.
Clonal propagation: asexual multiplication of plants from a single individual or
explant.
Clones: a group of plants propagated from vegetative parts, which have been derived
by repeated propagation from a single individual. Clones are considered to be
genetically uniform.
Contamination: infected by unwanted microorganisms in controlled environment.
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Cryopreservation: ultra-low temperature storage of cells, tissues, embryos and
seeds.
Culture: A plant growing in vitro in a sterile environment.
Differentiated: cultured cells that maintain all or much of the specialized structure
and function typical of the cell type in vivo.
Embryo culture: In vitro culture of isolated mature or immature embryos.
Explant: an excised piece or part of a plant used to initiate a tissue culture.
Ex vitro: Organisms removed from tissue culture and transplanted; generally plants to
soil or potting mixture.
Hormone: Generally naturally occurring chemicals that strongly affect plant growth.
In Vitro: To be grown in glass.
In Vivo: To be grown naturally.
Laminar Flow Hood: An enclosed work area where the air is cleaned using HEPA
filters.
HEPA= HIGH EFFIECENCY PARTICULATE AIR
Medium: a solid or liquid nutritive solution used for culturing cells.
Meristem: a group of undifferentiated cells situated at the tips of shoots, buds and
roots, which divide actively and give rise to tissue and organs.
Micropropagation: multiplication of plants from vegetative parts by using tissue
culture nutrient medium.
Propagule: a portion of an organism (shoot, leaf, callus, etc.) used for propagation.
Somatic embryos: non-zygotic bipolar embryo-like structures obtained from somatic
cells.
Subculture: the aseptic division and transfer of a culture or portion of that culture to a
fresh synthetic media.
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Totipotency: Capacity of plant cells to regenerate whole plants when cultured on
appropriate media.
Transgenic: plants that have a piece of foreign DNA.
Undifferentiated: cells that have not transformed into specialized tissues.
Reasons for in vitro cultivation of plant material:
Controlled environmental conditions
Science, symbiotic orchid seed culture, metabolite production
Pathogen-free material
Once pathogen-free, the material propagated under sterile conditions remains
pathogen-free
Multiplication
Rapid clonal propagation; also done ex vitro for many plants
Embryo rescue (infertile hybrids)
Cryopreservation
Propagation and Distribution of Mutants, Colourmorphs, etc.
Possible also for ex vitro clonally propagated plants
Genetic engineering
True regeneration necessary, most meristems are not transformable.
History of plant tissue culture:
The science of plant tissue culture takes its roots from the discovery of cell
followed by propounding of cell theory. In 1838, Schleiden and Schwann
proposed that cell is the basic structural unit of all living organisms. They
visualized that cell is capable of autonomy and therefore it should be
possible for each cell if given an environment to regenerate into whole
plant. Based on this premise, in 1902, a German physiologist, Gottlieb
Haberlandt for the first time attempted to culture isolated single palisade
cells from leaves in knop‘s salt solution enriched with sucrose. The cells
remained alive for up to one month, increased in size, accumulated starch
but failed to divide. Though he was unsuccessful but laid down the
foundation of tissue culture technology for which he is regarded as the
father of plant tissue culture. After that some of the landmark discoveries
took place in tissue culture which are summarized as under:
- 1902 - Haberlandt proposed concept of in vitro cell culture
- 1904 - Hannig cultured embryos from several cruciferous species
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- 1922 - Kolte and Robbins successfully cultured root and stem tips
respectively
- 1926 - Went discovered first plant growth hormone –Indole acetic acid
- 1934 - White introduced vitamin B as growth supplement in tissue culture
media for tomato root tip
- 1939 - Gautheret, White and Nobecourt established endless proliferation of callus
cultures
- 1941 - Overbeek was first to add coconut milk for cell division in Datura
- 1946 - Ball raised whole plants of Lupinus by shoot tip culture
- 1954 - Muir was first to break callus tissues into single cells
- 1955 - Skoog and Miller discovered kinetin as cell division hormone
- 1957 - Skoog and Miller gave concept of hormonal control (auxin: cytokinin) of organ
formation
- 1959 - Reinert and Steward regenerated embryos from callus clumps and cell
suspension of carrot (Daucus carota)
- 1960 - Cocking was first to isolate protoplast by enzymatic degradation of cell wall
Plant Tissue Culture: Current Status and Opportunities 3
- 1960 - Bergmann filtered cell suspension and isolated single cells by plating
- 1960 - Kanta and Maheshwari developed test tube fertilization technique
- 1962 - Murashige and Skoog developed MS medium with higher salt concentration
- 1964 - Guha and Maheshwari produced first haploid plants from pollen grains of
Datura (Anther culture)
- 1966 - Steward demonstrated totipotency by regenerating carrot plants from single
cells of tomato
- 1970 - Power et al. successfully achieved protoplast fusion
- 1971 - Takebe et al.regenerated000000 first plants from protoplasts
- 1972 - Carlson produced first interspecific hybrid of Nicotiana tabacum by protoplast
- 1974 – Rein hard introduced biotransformation in plant tissue cultures
- 1977 - Chilton et al. successfully integrated Ti plasmid DNA from Agrobacterium
tumefaciens in plants
- 1978- Melchers et al. carried out somatic hybridization of tomato and potato resulting
in pomato
- 1981- Larkin and Scowcroft introduced the term somaclonal variation
- 1983 - Pelletier et al.conducted intergeneric cytoplasmic hybridization in Radish and
Grape
- 1984 - Horsh et al. developed transgenic tobacco by transformation with
Agrobacterium
- 1987 - Klien et al. developed biolistic gene transfer method for plant transformation
- 2005 - Rice genome sequenced under International Rice Genome Sequencing
Project.
Terminology in plant tissue culture:
Callogenesis: Callus Formation
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Embryogenesis: Somatic Embryo Formation
Organogenesis, Caulogenesis: Shoot Formation
Rhizogenesis: Root Formation
Types of Culture.
Callus culture
Suspension culture
Embryo culture
Endosperm culture
Meristem culture
Protoplast culture
Stage of plant tissue culture:
Stage 0: Preparation of donor plant. Any plant tissue can be introduced in vitro.
To enhance the probability of success, the mother plant should be ex vitro cultivated
under optimal conditions to minimize contamination in the in vitro culture.
Stage I: Initiation stage.
In this stage an explant is surface sterilized and transferred into nutrient medium.
Generally, the combined application of bactericide and fungicide products is suggested.
The selection of products depends on the type of explant to be introduced. The surface
sterilization of explant in chemical solutions is an important step to remove
contaminants with minimal damage to plant cells. The most commonly used
disinfectants are sodium hypochlorite, calcium hypochlorite , ethanol and mercuric
chloride (HgCl2).The cultures are incubated in growth chamber either under light or dark
conditions according to the method of propagation.
Stage II: Multiplication stage.
The aim of this phase is to increase the number of propagules. The number of
propagules is multiplied by repeated subcultures until the desired (or planned) number
of plants is attained.
Stage III: Rooting stage.
The rooting stage may occur simultaneously in the same culture media used for
multiplication of the explants. However, in some cases it is necessary to change media,
including nutritional modification and growth regulator composition to induce rooting and
the development of strong root growth.
Stage IV: Acclimatization Stage.
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At this stage, the in vitro plants are weaned and hardened. Hardening is done
gradually from high to low humidity and from low light intensity to high light intensity.
The plants are then transferred to an appropriate substrate (sand, peat, compost etc.)
and gradually hardened under greenhouse.
IMPORTANCE OF TISSUE CULTURE:
To conserve rare or endangered plant species.
The commercial production of plants used as potting, landscape and florist
subjects, which uses meristem and shoot culture to produce large numbers of
identical individuals.
A plant breeder may use tissue culture to screen cells rather than plants for
advantageous characters, e.g. herbicides resistance/tolerance.
Large scale growth of plant cells in liquid culture in bioreactors for production of
valuable compounds, like plant-derived secondary metabolites and recombinant
proteins used as biopharmaceuticals.
To cross distantly related species by protoplast fusion and regeneration of the
novel hybrid.
To rapidly study the molecular basis for physiological, biochemical and
reproductive mechanisms in plants, for example in vitro selection for stress
tolerant plants.
To cross pollinate distantly related species and then tissue culture the resulting
embryo, this would otherwise normally die.
For chromosomes doubling and induction of polyploidy, for examples doubled
haploid, tetraploids and other forms of polyploids. This is usually achieved by
application of antimitotic agents such as colchicine or oryzalin.
As a tissue for transformation, followed by either short term testing of genetics
constructs or regeneration of transgenic plants.
Production of identical sterile hybrid species can be obtained.
Removal of viruses by propagation from meristem tissues.
Only a small explant is enough to get millions of plant with extremely high
multiplication rate.
Micropropagation:
Micropropagation starts with the selection of plant tissues (explant) from a
healthy, vigorous mother plant .Any part of the plant (leaf, apical meristem, bud and
root) can be used as explant.
Embryogenesis:
The process of initiation and development of embryos or embryo-like structures from
somatic cells (Somatic embryogenesis).
Organogenesis:
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The process of initiation and development of a structure that shows natural organ form
and/or function.
Factors determining success with plants TC.
Status of donor plant.
•Species, genotype, age of tissue, explant size and type.
Experimental conditions.
•Temperature, light, day/night.
Composition of culture medium.
Important:
All factors are genotype-dependent and require optimization for each cultivar.
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PATHWAY OF TISSUE CULTURE
Morphogenesis:
Morphogenesis is the biological process that causes an organism to develop its
shape. Morphogenesis can take place also in a mature organism, in cell culture
or inside tumor cell masses.
↓ ↓
PATHWAY OF MORPHOGENESIS
Direct Morphogenesis
Rooting Under in-Vitro
Plant
Regeneration of Plants From
Callus
Explant
Indirect Morphogenesis
Explant
Shoots
Rooting Under in-Vitro
Plant
Hardening
Plantlets
Hardening
Plant
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CULTURE MEDIA
A growth medium or culture media is a semi solid designed to support the growth of
microorganisms or sell or small plants. There are different types of medium for growing
different types of cells or plant.
Types of media:
Synthetic media :
When a medium is composed of chemically defined components, it is referred to as
synthetic media.
Natural media:
If a medium contains chemically undefined compounds ( e.g. vegetable extract, fruit
juice, plant extract), it is regarded as a natural media.
Commonly used media:-
White’s medium:
This is one of the earliest plant culture media developed for root culture.
MS medium :
Murashige and skoog (MS) originally formulated a medium to induce organogenesis,
and regeneration of plants in cultured tissue. These days, MS medium is widely used for
many types of culture system.
B5 Medium :
Developed by Gamborg, B5 medium was originally designed for cell suspension and
callus culture. Presently used for protoplast culture.
N6 Medium:
Chu formulated this medium and it is used for cereal anther culture, beside other tissue
culture.
Nitsch’s Medium:
This medium was developed by Nitsch and Nitchs and frequently used for anther
culture.
Culture Medium constituents:
• Inorganic salt formulations.
• Source of carbohydrate.
• Vitamins.
• Water.
• Plant hormones - auxins, cytokinins, GA‘3.
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• Solidifying agents.
Plant tissue culture media:
Inorganic compounds (mineral nutrition)
Carbohydrates (typically sucrose)
Plant Growth Regulators (PGRs) (hormones)
Miscellaneous compounds
Plant tissues cultured in vitro require a balanced supply of nutrients.
Essential Nutrients:
Inorganic compounds.
Macronutrients.
•N, P, K, Mg, Ca, S.
Micronutrients.
•Mn, I, Cu, Co, B, Mo, Fe, Zn, (Ni, Al).
Mineral salt composition:
Macroelements: The elements required in concentration > 0.5 mmol/l.
The essential macroelements: N, K, P, Ca, S, Mg, Cl.
Microelements: The elements required in conc. < 0.5 mmol/l.
The essential microelements: Fe, Mn, B, Cu, Zn, I, Mo, Co.
The optimum concentration → maximum growth rate.
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Mineral salts:
Function of nutrients in plant growth:
Plant growth regulators:
(Body building Plants)
Auxins:
- induces cell division, cell elongation, swelling of tissues, formation of callus, formation
of adventitious roots.
- inhibits adventitious and axillary shoot formation
- 2,4-D,
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-NAA,
-IAA,
-IBA,
-pCPA
Cytokinins:
- shoot induction, cell division.
- BAP,
-Kinetin,
-Zeatin,
-2iP
Gibberellins:
Plant regeneration, elongation of internodes
- GA3…
Abscisic acid:
Induction of embryogenesis
- ABA
Cultured tissue must contain competent cells or cells capable of regaining competence
(dedifferentiation). e.g. an excised piece of differentiated tissue or organ (Explant)
→dedifferentiation → callus (heterogenous) → redifferentiation (whole plant) = cellular
totipotency.
1957 Skoog and Miller demonstrated that two hormones affect explants’
differentiation:
– Auxin: Stimulates root development
– Cytokinin: Stimulates shoot development
• Generally, the ratio of these two hormones can determine
plant development:
– ↑ Auxin ↓Cytokinin = Root development
– ↑ Cytokinin ↓Auxin = Shoot development
– Auxin = Cytokinin = Callus development
Typical plant growth regulators and their effects on tissue cultures:
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Auxins →
•callus
•roots
Cytokinins → •shoots
•embryoids
Gibberellic acids →
•cell growth
•elongation
Auxins:
Plant growth and physiological functions.
Phototropism .
Apical dominance.
Cell division.
Differentiation .
Initiation of embryos, organs (esp. roots).
Synthetic auxins are often more effective than the natural auxins.
Cytokinins:
Main effects in tissue culture systems.
Adventitious shoot formation (at high conc.).
Inhibition of root formation.
Cell division.
Callus formation and growth.
Stimulation and outgrowth of axillary buds.
Inhibition of leaf senescence.
Gibberellic acid (GA):
Synthesized irom mevalonate .
Shoot and root apices, embryos, cotyledons, fruit, tubers .
Only some forms are biologically active: GA3.
Dramatic effects on cell elongation.
Promotes cell division in combination with IAA.
Effects on seed germination (breaking seed dormancy).
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Improves fruit set, fruit growth, fruit maturation and fruit ripening.
Promotes flowering.
Solidifying Agents:
For the preparation of semisolid or solid tissue culture media, solidifying or gelling
agents are required. In fact, solidifying agent extend support to tissue growing in the
static conditions.
Agar: A polysaccharide obtained from seaweeds, is most commonly used as a
gelling agent for the following reasons:
(a) It does not react with media constituents.
(b) It is not digested by plant enzymes and is stable at culture temperature. Agar at a
concentration of 0.5 to 1% in the medium can form a gel.
(c)
Gelatin: It is used at high concentration (10%) with a limited success. This is
mainly because gelatin melts at low temperature (25%) and consequently the
gelling property is lost.
Other Gelling Agents: Bio-gel (polyacrylamide pellets), Phytagel, gelrite an d
purified agrose are other solidifying agents, although less frequently used.it is in fact
advantageous to use synthetic gelling compounds, since they can form gels at a
relatively low concentration (0.1 to 2.5 gl-1
)
pH of Medium: the optimal pH for most tissue cultures is in range of 0.5-0.6. At a pH
higher than 7.0 and lower than 4.5, the plant cells stop growing in culture. If general, pH
above 6.0 give the medium hard appearance, while below 5.0 does not allow gelling of
the medium.
Nutrient medium pH range of 5.7 to 5.8 is suitable in-vitro growth of explant.
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INSTRUMENTATION:
Following instruments were used in SFRI during study:
a. Types of Autoclave:
1. Horizontal autoclave.
2. Vertical autoclave.
1. Horizontal autoclave:
It was used for discarding contaminated cultures. Contaminated culture bottles,
test tubes etc. were kept one hour at 15 PSI pressure and 121o
C temperature in
autoclave.
Fig:Horizontal autoclave
2. Vertical autoclave:
Used for sterilization of media and glassware‘s. Media was kept for 20
minutes and glassware‘s was kept 1 hour at 15 PSI and 121o
C temperature
for purpose of sterilization.
Fig: Vertical autoclave
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b. RO Water Purifier:
The distillation process removes minerals and can reduce levels of chemical
contaminations.
A water distiller works by boiling water into water vapour, condensing it and
returning it to its liquid state. It is collection in a storage container.
Fig: RO Water Purifier
c. Microwave oven :
A Microwave oven was used to melt microbiological media, resulting in a
substantial reduction of heat generation and considerable saving time.
Fig: Microwave oven
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26
d. PH Meter and Magnetic Stirrer:
They are used for maintaining the pH of the solution and media. pH of the
media should b in the range of 5.7 to 5.8, which is adjusted by adding 1N HCL
or NaOH.
Magnetic stirrer was used to stir and mix solutions for precise periods, from
short as a minute to long as a few days.
Fig: pH Meter and Magnetic Stirrer
e. Electronic Balance:
It is used to weight accurate amount of salts and chemicals for media
preparation.
Fig:Electronic Balance
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27
f. Hot Air Oven:
It is used for sterilized of glassware‘s at 60 o
C for one day and 180 o
C for one
hour. Its temperature ranges from 25 o
C to 300 o
C.
Fig: Hot Air Oven
g. Laminar Air Flow:
They are used for inoculation purpose. Before inoculation LAF cabinet was
wiped with 70% alchol and then exposed to UV Light for 40 minutes.
Fig:Laminar Air Flow
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At the early stages of planning it is important to ensure that the basic infrastructure such
as continuity of power and supply of appropriate quantity of water are in place that local
back-up support service are available for the repair and servicing of equipment‘s. a
standard plant tissue culture laboratory should have facility for:
Washing and storage of glassware, plastic ware and lab ware.
Preparation, sterilization and storage of nutrient media.
Aseptic manipulation of plant material.
Maintenance of culture under controlled condition of light and temperature.
Greenhouse for hardening of plantlets.
HPLC unit for separation of sample.
a. Washing room:
Should be provided with appropriate washing are, running tap water, brushes
of various shapes and sizes, racks.
It may also have a dust proof cabinet to store them.
Fig: Washing room
b. Sterilization room:
Should have hot air oven for drying glassware/ lab wares and sterilization of
lab equipment‘s.
It should also have autoclave for sterilization of glassware‘s, distilled water and
media.
Fig: Sterilization room
Lab infrastructure
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c. Media preparation room:
Should have shelves for various chemicals required, refrigerator to store
chemicals, Electronic balance and benches for working.
Also provided with hot-plate cum magnetic stirrer, pH meter and microware
Oven.
Fig: Media preparation room
d. Inoculation room:
It should be provided with LAF (Laminar Air Flow) for aseptic manipulation of
explants
It have table for keeping media, sterilized cotton. Air conditioner and SDDW
(sterilized double distilled water)
Fig: Inoculation room
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e. Culture room:
It should be provided with controlled condition of light (1000 lux) and air
conditioners to maintain temperature around 25 to -2o
C.
Fig: Culture room
f. Mist chamber :
It should be provided with proper green shed.
Also provided with proper water provision and only fertilized soil is used for
Hardening of plant.
Fig: Mist chamber
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GLASSWARE’S AND PLASTIC WARE WASHING
A detergent (Labolene) especially designed for washing are available. Washing is done
in fallowing steps:-
Overnight soaking of glassware‘s or labwares in detergent solution.
Thoroughly rinsing first with tap water and then with distilled water.
Drying in hot air oven at 70_75o
for 1 hour.
Storing in dusting proof cupboards.
Sterilization Techniques:
Media, glassware‘s or plastic wares, instruments and plant material (explant)
are sterilized to keep them free from microbes i.e. to maintain aseptic
condition.
The instructions or equipment‘s used for aseptic manipulation such as forceps,
scissors, scalpels, needles and spatula are normally sterilized by dipping in
95% ethanol followed by flaming and cooling. This is done at the start of the
transfer or inoculation work and several times during operation to minimize the
contamination.
Techniques:
S.No Sterilization techniques Material sterilized
1 Physical method.
Moist heat (autoclaving) (121o
C at 15
PSI for 20 to 40 minutes).
Media, culture vessels glassware‘s
or plastic wares and Contamined
cultures.
2 Dry heat (160o
C to 180o
C for 1 hours). Empty glassware‘s like culture
bottles, pipettes, measuring cylinder,
test tubes etc.
3 Flaming (red hot). Needle. Scalpels, Forceps and
Scissors.
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Bhaans (Kashmir); Jati/Mirtinga/Wat (Arunachal Pradesh); Jati (Assam); Owati
(Meghalaya); Koraincho bans (Sikkim); Longmeii (Ao –Nagaland); Rawthing (Mizoram);
Mirtinga (Tripura).
Habitat:
This species occurs at an altitude of 1500 m. it prefers moist alluvial soil in good rainfall
areas and fine textured soil in semi evergreen forest, in relatively low rainfall areas with
subtropical to temperate climatic condition.
Distribution:
Distributed widely in North Eastern India and West Bengal.
Flowering and fruiting:
This species flower gregariously. The flowering cycle is 30 – to 60 years.
Identification features:
A large tufted bamboo.
Culm up to 20m high and 8 cm in diameter, smooth; internodes 40-70 cm long.
Culm sheath 20-25 cm long and broad, nearly glabrous, rounded at tip, black inside;
blade 10-15 cm long, triangular, cuspidate, appressed hairy beneath, rounded at base;
ligule 2 mm high, white hairy outside.
Leaves 20-35 cm long and 3-4 cm broad, oblong-lanceolate, base oblique, petiole
short; leaf-sheath glabrous or sparsely hairy, ligule short.
Silvicultural management techniques:
The seeds exhibit orthodox behavior and can be stored by proper control of moisture
content and temperature. Studies on seed viability shows that under natural condition
the seed are viable not more than two months but this can be extended by storing over
anhydrous silica gel in desiccators up to 18 months.
Vegetative propagation like rhizome and culm cutting are successfully practiced for
propagation of this species apart from seeds. The seedlings raised from culm cuttings
can be successfully multiplied by shoot proliferation. As per felling rules, felling cycle
Local name
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four years is suggested. Culm less than one year should be retained and cuttings
should be made 30 cm above the ground. A minimum of six culms should be left in a
clump. This species is one of the high yielding bamboos suggested for large scale
plantation.
Pest and diseases and their control:
The sap sucker Oregma bumbusae which causes the wilting and death of young shoots
have been reported. Bavistin, BHC powder or dialdrin or aldrin (0.5 percent solution or
powder) are affective control. Fungal infection also affects the yield and quality of pulp.
The species also affected by blight caused by Sarocladium oryzae. This can be
controlled by cultural practice and application of Dithane M45as soil drench.
Uses:
It is favored for handicraft, paper and structural purpose. It is a strong bamboo; it lends
itself easily to mechanized processing, and is being used for making bamboo boards
and composites.
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REVIEW OF LITERATURE
Pratibha Sharma and K.P Sarma. In vitro auxiliary shoot formation was highest in
Murashige and Skoog basal medium supplemented with 1.0 mg/l 6-Benzyle Adenine
(BA). Subcultures of shoots from clumps were continued several times for maintaining a
stock of mother culture. Clumps of at least 3 shoots were used for root induction in MS
medium with Indole-3- Acetic Acid (IAA), Indole-3- Butyric Acid (IBA) and naphthalene
acetic acid (NAA).Response of rooting was found more in 5.0 mg/l naphthalene acetic
acid. Rooted plantlets were successfully acclimatized in green house for 20 to 25 days
and then were transferred to the natural field condition. The survival rate was recorded
100 percent in field condition. To the best of our knowledge this is the first report on in
vitro generation of Bambusa tulda from mature field grown auxiliary bud in commercial
scale in North-East India. In this paper, a continuous mass multiplication protocol of B.
tulda was described, which is cost effective, easy to raise, economic to adopt, easy to
transport for selling purpose
In B.tulda, Banik (1987b) was able to increase the seed longevity period up to 18
months by storing over silica gel in a desiccator. Efforts to prolong the viability of fleshy
recalcitrant bamboo seeds by conventional storage methods were not promising.
Reports show that storing the orthodox seeds of bamboos over calcium chloride with a
moisture content of 10-11 % is ideal. The viability of seeds of B. bambos and B. tulda
was extended by storing the seeds over calcium chloride at room temperature.
Seedlings grow well in partial shade compared to direct sunlight. The germinating
plumules are very thin in B. tulda and thick in M.baccifera. Within 1-4 weeks, plumules
elongate rapidly into stems bearing single leaves arising alternately. The stems of B.
tulda, B.longispathus, and B. polymorpha are more or less woody in nature,but M.
baccifera has a soft and succulent stem with vigorous growth. Arhizome system starts
to develop in the seedling one or two monthsafter germination.
Reported that 5-9 month old seedlings of B. tulda can be multiplied 3-5 times in number
through this technique. Every year the seedling can be multiplied at the same rate,
keeping a stock for future macro proliferation. The survival rate of these multiplied
seedlings is 90-100%.
Macro-proliferation, a method of plant multiplication by separating the rooted tillers has
been used by many workers for enhancing the rate of multiplication of in vitro raised
plants and for continuous supply of plantlets. Splitting of rooted tillers could double the
production of Dendrocalamus asper plants (Singh et al., 2011) while three-fold increase
was achieved in and Bambusa balcooa (Mudoi and Borthakur 2009), .B tulda (Mishra et
al., 2011)
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Mishra Y, Patel PK, Yadav S, Shirin F, Ansari SA (2008). A micropropagation system
for cloning of Bambusa tulda Roxb. Sci. Hortic. 115:315-318.
Yogeshwar Mishra, Pradeep Kumar Patel, Suman Yadav, Fatima Shirin, S.A.
Ansari.
The communication describes standardization of an efficient in vitro propagation and
hardening procedure for obtaining plantlets from field grown culms of Bambusa tulda.
Administration for 10 min of 0.05 and 0.1% mercuric chloride to explants collected in
winter and summer seasons, respectively facilitated optimum culture establishment and
bud break. 0.1–0.2% mercuric chloride in rainy season enhanced aseptic culture
establishment but inhibited bud break due to toxicity to explants. MS liquid medium
enriched with 100 mMglutamine, 0.1 mMindole-3-acetic acid and 12 mM 6-
benzylaminopurine supported maximum in vitro shoot multiplication rate of two-fold. The
proliferated shoots were successfully rooted on MS liquid medium supplemented with
40 mM coumarin resulting in a maximum of 98% rooting. The procedure requires 45
days cycle for the in vitro clonal propagation (15 days for shoot multiplication and 30
days for root induction) and 80 days for acclimatized plantlet production.
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CoCl2.6H2O 5
Stock-III
(200x)
Iron
FeSo4.7H2O 5560
Na2EDTA.2H2O 7460
Stock-IV
(200x)
Vitamins
Inositol 20000
Nicotinic acid 100
Pyridoxine Hcl 100
Thiamine 20 mg/l
Glycine 400
Volume of stock 2000 ml 1000 ml 500 ml
Stock I 100 ml 50 ml 25 ml
Stock ii 10 ml 5 ml 2.5 ml
Stock iii 10 ml 5 ml 2.5 ml
Stock iv 10 ml 5 ml 2.5 ml`
PGR (Plant Growth Hormone) As per need.
Sucrose 3% (30 g/l)
pH 5.7 -5.8 using 1 Hcl or NaOH
Agar 0.8% (8 g/l)
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41
Take 500 ml DDW in a conical flask
Add 8 gm Agar to the solution
Adjust the pH of the solution 5.7-5.8with the pH meter
Now add required amount of PGR
Add 5 ml of Stock-II and stir
Then 5 ml of Stock-IV and shake well
Add 50 ml of Stock-I and stir
Add 5 ml of Stock-III and stir
Add 30 gm Sucrose and dissolve it
Now pour the media in test tubes or bottles
Keep it in the microwave to melt the Agar
Sterilize them in autoclave at 121oC at 15 PSI pressure for 30 minutes
MS Media Preparation
For 1 Litre Media
Important:
Auxin dissolved in alcohol.
Cytokines dissolved in NaOH.
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42
Explants were collected in bottle early in morning
Explant were cleaned with Alcohol and cut required parts
Cover the top of bottle with muslin cloth
Cover the top of bottle with muslin cloth
Wash the explant with DDW for (3-4) times
Wash the explant with DDW for (3-4) times
Wash the explant with Extren detergent
Now washing with 1% Bavistin (Fungicide) for 10 minutes
Keep with under running tap water for 5 minutes
Again wash the explant with DDW for 3-4 times
COLLECTION OF
EXPLANT & WASHING
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WASHING & INOCULATION OF EXPLANT IN LAMINAR AIR
FLOW
Washing the Explants with SDDW for 3-4 times
Labelled with the name of explant, date of inoculation
Now cover the test tubes / bottles along with cap by ceiling tape
Now innoculate the Explant in front of flame and cover the cap tightly
Put the Explants in petriplates and allow to dry
Again wash the Explants with SDDW for 3-4 times
Now wash the Explants with 0.1% (100 mg) HgCl2 for 2 minutes
Finally put the culture bottles or test tubes in culture room under the
proper light and temperature
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↓
↓
↓
↓
↓
PREPARATION AND WASHING OF LAF
Wipe out LAF with 70% alcohol
Put all the requirements and bottles of SDDW and a switch on UV light for
1 HOUR
Expose the LAF Cabinet under UV light for 1 HOUR
Again wipe out LAF, test tubes/bottles containing media with Alcohol
Dip scalpel and forceps into alcohol containing tubes
Wipe out hands with 70% Alcohol
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↓
↓
↓
↓
↓
↓
↓
TECHNIQUE FOR SUBCULTURING
Wipe out LAF 70% alcohol and expose the LAF cabinet with UV light
for 30 minutes
Now put all the requirements including media bottles and fresh
culture tubes and expose to UV for 30 minutes
Wipe out hands and LAF surface with 70% alcohol before use
Forcep, scalpel and petriplates were wiped out with alcohol and flame
sterilized
Put the explants in petriplates and slowly remove the dry part and
media stick to explant to avoid contamination
Properly cut the lower and upper part of explant to remove dead cells
and carefully inoculation the explant in media bottle
Bottles were labeled with name of explant, date of subculturing and
cover the top of bottle tape
Finally put the culture bottles in culture room under the proper light
and temperature
OBSERVATION OF CULTURE VESSELS:
Culture tubes / bottles were timely observed for
growth and contamination.
Contaminated cultures were immediately
removed.
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47
EFFECT OF DIFFERENT TIME INTERVAL AS WELL AS CONCENTRATION
AND TREATMENT DURING FRESH CULTURING:
NAME of EXPLANT - BAMBUSA TULDA
PGR
Combination
TREAMENT
% OF RESPOND SHOOTS CONTAMINATION %
SHOOTS
Avg. data
I
week
II
week
III
week
I
week
II
week
III
week
I week II week
III week
No
Length
(cm)
No
Length
(cm)
No Length
(cm)
CONTROL
RT-5min
93% 86% 13% 13%
2.2
EXTRIN-2gm
5 min
BAVISTIN-
2gm, 15 min
86% Nil 1.3 1.2 2.8
3.1 3.0
Hgcl2-0.1% 4
min
1BAP
1mg/l
RT-5min
EXTRIN-2gm
5 min
BAVISTIN-
2gm 15 min
Hgcl2-0.1% 5
min
100% 100% 90% Nil Nil 10% 1.6 1.5 2.6 3.1 3.2 3.5
RT=Running Water
BAP=benzyl 6-amino purine.
Maximum number of shoot multiplication and their better respond was observed at the
concentration and combination of 1.0 mg/l Benzyl Amino Purine.
In control there is a less number of shoot multiplication.
From the above table it is recommended that during fresh culturing the effect of Hgcl2
was found to best with 0.1 Hgcl2 concentration for 5 minutes as compared to 4 minutes.
The species of Bambusa tulda showed best response in the PGR combination of 1.0
BAP 1mg/l. so the subculturing of the explants was under process in the same
combination of PGR.
Observation table
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Images Showing The Growth Of
Explant At Different Time Intervals
Figure 1:-Growth after 7 days in 1 mg/l BAP)
Gr
Figure 1:-Growth after 14 days in 1 mg/l BAP)
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Conclusion
The present study was conducted at the plant tissue laboratory, deparment of Forest
Genetic, Plant Propagation & Biotechnology Division,state Forest Research Institute
Jabalpur Madhya Pradesh, during the period from 30 may to 28 june to propagate from
combinations of the control and PGR medium for micropopagation of Bumbusa tulda.
Murashing and skoog medium was used.
The nodal cutting explants, maximum number of shoot multiplication and their better
respond was observed at the concentration and combination of 1.0 mg/ Benzyl Amino
Purine optimum 3.2 shoot with 3.5 cm length after 21 IIIrd week.+
53. SFRI
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Our experiences in SFRI Jabalpur Madhya Pradesh
About wildlife
Mr. R.Bisen (ACF)
Wildlife:
Wildlife is all non-domesticated plants, animals, and other living things.
Domestication, act of taming, or controlling, wild plant and animal species
and producing them for human benefit, is performed often and has an
impact on the environment, both positive and negative.
Wildlife Conservation:
Wildlife conservation is the practice of protecting endangered plant
and animal species and their habitats.
Among the goals of wildlife conservation are to ensure that nature
will be around for future generations to enjoy and to recognize the
importance of wildlife and wilderness lands to humans.
Many nations have government agencies dedicated to wildlife
conservation, which help to implement policies designed to protect
wildlife. Numerous independent nonprofit organizations also
promote various wildlife conservation causes.
Wildlife conservation has become an increasingly important
practice due to the negative effects of human activity on wildlife.
The science of extinction is called dirology.
An endangered species is defined as a population of a living being
that is at the danger of becoming extinct because of several
reasons.
Either they are few in number or are threatened by the varying
environmental or predation parameters.
Major threats to wildlife
Habitat loss—due to destruction
Habitat fragmentation
Habitat degradation: pollution, invasive species
Climate change
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Unregulated Hunting and poaching
Over exploitation
Wildlife values.
Positive values
Tangible Intangible
Negative values:
Wildlife damage.
Man-animals conflict.
Economic loss.
Physical utility
Economic value
Recreational value
Historical value
Scientific value
Ecological value
Existence value
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Positive values Tangible:
Physical utility: as food, clothing and others as domestic
Economic value: furs, hides, every and medicines
Recreational value: NP, sanctuary, bird watching, tourism
Historical value: About historical knowledge of wildlife.
Intangible
Scientific value: research and development of new things.
Ecological value: maintaining ecological balance.
Existence value: future potentiality that helps in preservation of
genetic diversity.
Negative values
Wildlife damage: damage to agriculture as well as forest crops.
Man-animals conflict: human death, injury and illness and disease
chikungunea.
Economic loss: reduced the productivity of forest and agriculture
crop.
PRINCIPLES OF CONSERVATION
The management of biosphere to yield sustainable benefit to present
generations, maintaining its potential to meet the needs and aspirations of
future generations‖ is called as conservation. there is an urgent need to
conserve the biodiversity. The conservation of biodiversity has the following
objectives:
To preserve the biodiversity.
Maintenance of ecological balance.
Sustainable utilization of resources for all.
METHODS OF CONSERVATION
In situ conservation: ―Conservation of species in its natural habitat or
ecosystem. It involves setting large areas for wildlife conservation and
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protection of endangered species. There are different categories of
protected areas. They are
1> National Parks,
2> Sanctuaries,
3> Biosphere Reserves,
4> Protected forests
1. National Parks
A relatively large area which is strictly reserved for the conservation of wild
life forms. Deforestation, grazing, cultivation and private ownership are not
permitted in this area. These are designated to conserve specific wild
animals like Tiger, Lion etc. along with other life forms. The parks are
around 100-1000sq.km. and boundaries are circumscribed by legislation.
Except buffer zone, no biotic interference and tourism permissible. There
are 90 national parks are occur in India and some of them are listed
below.
SV National Park – Andhra Pradesh – Leopard, Elephant etc.
Gir NP – Madhya Pradesh – Lions, Antilopes
Khaziranga National Park – Assam – Rhinoceros, Leopard etc.
Sundarbans – West Bengal - Tigers
2. Sanctuaries.
These are meant for conservation of specific animal and plant. The area is
about 100-1000sq.km and there is no legislative boundaries. Grazing,
timber harvesting, collection of NTFP (Non Timber Forest Products) and
private ownership are accepted. There are 492 wildlife sanctuaries in India.
Rajeev Gandhi wildlife sanctuary of Andhra Pradesh is meant for
conservation of Tiger, Leopard and Crocodiles. Madhumalai wildlife
sanctuary (Tamil Nadu) is meant for conservation of Elephant, Four horned
Antelopes, Kumbhalgarh wildlife sanctuary in Rajasmand district Rajasthan
for bears, Sitamata wildlife sanctuary for flying squirrel, Bheinsroadgarh
wildlife sanctuary for leopard.
3. Biosphere reserves
These are meant for protecting and conservation of entire ecosystem.
First time UNESCO (United Nations Education Social & Cultural
Organization) proposed Biosphere reserves in 1972 through Man &
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Biosphere Program . Local tribal people would be allowed to continue
their traditions but have no specific legal status. Commercial activities
and construction of projects not allowed in Biosphere reserves. It
have Core zone, Natural disturbed area, Manipulation zone. . In India
18 Biosphere Reserves are occurred,
4. Protected forests
A protected forest is a forest with some amount of legal or constitutional
protection, or where the habitat and resident species are legally accorded
protection from further deplication .
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Forest mensuration
Mr. SK Jain (Acf)
Introduction: It is an essential part of mathematics. The word is derived
from latin word ‗mensura‘ which means ‗measure‗.
Mensuration is a branch of forestry which deals with the determination of
dimensions (e.g. diameter, height, volume etc), form, age and increment of
single tree, stands or whole woods, either standing or after felling
.(Chaturvedi and Khanna, 1982)
Objectives
Basis for sale.
Basis of management.
Measurement for research.
Measurement for planning.
Scope
Provides foundations of measurement principles applicable to any
forest measurement problems.
Application of statistical theory and use of computer for data
processing
Forecast of future yields.
Measurement not only standing trees but also felled timber and
their conversion.
Unit of measurement in forest measurement
1 mile=1.609km.
1 cft=0.0283 cu m.
1 cft/acre= 0.070 cu m/ha.
1 ha= 2.47105 acres.
1 cu m/ha= 14.291 cft/acre.
1 cubic metre= 35.3147 cubic ft.
1 kilogram= 2.20462 pounds.
1 metric=0.98420 ton.
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Measurement of Trees
The main objective of measurement of individual trees is to estimate the
quantity of timber, firewood or any other forest produce which can be
obtained from them. It covers:
Diameter and Girth Measurement.
Height Measurement.
Measurement of Logs and Fuel wood.
Diameter Measurement of Trees
The linear measurement, the main objective of which is to estimate
the volume of the trees.
The volume of a tree is dependent on diameter or girth at breast-
height, total height and form factor.
It is not only necessary for calculation of volume of logs, but also
necessary for making inventory of growing stock as well as to
correlate height, volume, age, increment of trees.
DBH Measurement
DBH is simply the average stem diameter outside bark at point,
1.37m above ground.
Universally adopted standard height for measuring girth, diameters
and basal area of standing trees India, Burma, America, Union of
South Africa and other British Colonies-1.37m.
In Europe, U.K., DBH is taken as 1.3m. It is recommended by FAO
as standard.
Significance of DBH
Convenient height for taking measurement.
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Avoids the fatigue caused unnecessarily.
Saves extra expenditure from not clearing the base.
Abnormalities, eg. Root swell, disappear below breast-height.
Standardizes diameter measurement giving a uniform point of
measurement. Diameter measurement at stump height is preferred,
but standardization is lost because height of stump depends upon
skill of the labor and the commercial value of the tree.
Points to be considered at the time of measuring diameter
Breast height point should be marked by intersecting vertical and
horizontal lines 12 cm long, painted with white paint.
Breast height should be marked by means of a measuring stick on
standing 1.37m or 4 ft 6 in above the ground level, but 1.3 (4‘3‖) in
case of FAO.
On the sloppy ground, the diameter at breast height should be
measured on the uphill side, after removing any dead leaves or
needles lodged there.
In the case of leaning tree, dbh is measured alone the tree stem
and not vertically.
Breast height mark should be shifted up or down as little as
possible to a more normal position of the stem and then diameter
measured if stem is abnormal.
Buttress is formed due to edaphic factor so if buttress is seen, the
dbh should be taken a little above the buttress formed.
Instruments for DBH measurement
Mostly used to measure dbh/girth in developing countries are,
Wooden scale.
Callipers
Tape.
Biltmore stick.
Penta prism.
Their use depends upon the condition of trees (felled or standing)
and the degree of accuracy required (research, business, etc.).
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Instruments for Height Measurement
1. Based on Similar Triangles.
Christen Hypsometer.
Smythies‘ Hypsometer.
Disadvantages
It‘s very slow, fatigue, heavy and rough information.
Advantages
Able to make manually even in the field
Used by unskilled labor.
2. Based on Trigonometrical Principles
Abney‘s Level.
Brandis Hypsometer.
Relaskop.
Haga Altimeter.
Disadvantages
Manufactured scientifically, repair is difficult on spot, Limited use
and expensive.
Only used by skilled labor.
Advantages
It‘s fast, easy to carry and accuracy maintained available in
markets
Adopted by many countries.
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