0.1 plant tissue culture step by step guide
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Selection of plant species Selection of mother plant for collection of explant Explant Surface sterilization Media composition for various objectives (Callus, somatic embryo, shoot etc) Genetic fidelity Acclimatization (Primary hardening, secondary hardening, field transfer)
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Plant tissue culture involves growing plant cells, tissues, or organs in a sterile environment with nutrient media. Some key events in the history of plant tissue culture include Haberlandt's proposal in 1902 that single plant cells could be cultured, White's work in the 1930s on growth regulators, and the discovery of kinetin by Miller and Skoog in 1957. Techniques for genetic transformation include protoplast fusion, using Agrobacterium tumerfaciens and its Ti plasmids, microinjection, and electroporation. Plant tissue culture is now used for cloning plants, introducing new traits through genetic engineering, and mass producing commercial crops.
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Plant tissue culture techniques are used to grow plant cells, tissues, or organs in sterile conditions. This allows for cloning of plants through micropropagation, where a small piece of plant tissue is grown in a nutrient medium, allowing for mass propagation. The document discusses the history and development of tissue culture, including early experiments in the 1800s and discoveries of growth hormones like auxin and cytokinin that allowed cultured cells and tissues to proliferate. It also outlines the stages of micropropagation including establishment, multiplication, root formation, and acclimatization of plants.
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The science of plant tissue culture takes its roots from the discovery of Cells (Robert Hooke in 1665) and 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 plants.
2. Plant Tissue Culture: Past & Present Prospects
In 1902, a German physiologist, Gottieb Haberlandt for the first time attempted to culture isolated single palisade cells from leaves in knop’s salt solution.
The cell remained alive for up to 1 month, increased in size, accumulated starch but failed to divide.
Though he was unsuccessful but he laid the foundation of tissue culture so he is regarded as Father of Plant Tissue Culture.
In the Subsequent years different landmark discoveries were made. Some of them are:
Use of specialized media for aseptic culture of Orchid seeds (Knudson, 1925) and other workers also demonstrated that plants could be propagated in vitro from the minuscule seeds of the Orchidaceae.
Further culture of other plant tissue was not possible due to lack of knowledge of the specific hormones to be added to the culture media.
This limitation was overcomed by the elucidation of the nature of Auxin, IAA, by Thimann and Went(1930) that plants would be subsequently regenerated through the use of IAA or its analogs.
Discovery of Cytokinins, specially Kinetin(6-furfurylaminopurine) by Miller et al. (1956), the regeneration of intact plants from tissue of many herbaceous species became a practical reality.
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Plant tissue culture media provides nutrients and environment for plant growth in vitro. It contains inorganic salts that provide minerals, organic compounds like sugars and vitamins, and gelling agents. Common media include Murashige and Skoog, Gamborg's B5, and Linsmaier-Skoog. These supply essential elements like nitrogen, phosphorus, and potassium as well as trace elements needed for plant development. Media is prepared by adding ingredients to water, adjusting pH, adding gelling agent, sterilizing by autoclaving, and allowing it to solidify before use in culture.
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This document provides a history of major developments in plant tissue culture from 1902 to 2005. Some of the key events include:
- 1902: Haberlandt proposed the concept of in vitro cell culture.
- 1954: Muir was the first to break callus tissues into single cells.
- 1955: Skoog and Miller discovered kinetin as a cell division hormone.
- 1960: Cocking was the first to isolate protoplasts by enzymatic degradation of cell walls.
- 1964: Guha and Maheshwari produced the first haploid plants from pollen grains through anther culture.
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Secondary metabolites are organic compounds produced by plants and organisms that are not essential for growth or reproduction. They play important roles in defense against herbivores and pathogens. Approximately 1500 new secondary metabolite molecules are identified from plants each year, around 30% of which show some biological activity. Secondary metabolites have many applications in medicine, food, cosmetics, and other industries. Plant tissue culture is used to produce many important secondary metabolites in a controlled environment, as production from native plants can be limited by environmental and geographical factors. Common production methods include cell suspension cultures, hairy root cultures, and immobilized cell cultures. Factors like media composition, temperature, pH, and elicitors can influence metabolite yield. An example is the production
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Introduction
Advantages of Micropropagation over the conventional methods
History
Stages of Micropropagation
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2. Stage 1; Initiation of aseptic cultures
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Factors affecting initiation stage
Conclusions
References
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The document discusses micropropagation, which is a type of tissue culture used to rapidly produce many uniform plants from small plant parts like meristems. It involves four stages: establishing an aseptic culture, multiplying propagules in nutrient media, pre-transplant rooting, and transplanting to soil. Micropropagation has advantages like producing many clean, genetically identical plants quickly and independently of seasons. It requires specialized facilities and expertise. Potential problems include contamination, tissue browning, abnormal growth, and shock from in vitro to in vivo conditions. Micropropagation has applications for rapidly increasing stocks of superior varieties and producing disease-free plants.
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Plant tissue culture involves growing plant cells, tissues, or organs in a sterile environment with nutrient media. Some key events in the history of plant tissue culture include Haberlandt's proposal in 1902 that single plant cells could be cultured, White's work in the 1930s on growth regulators, and the discovery of kinetin by Miller and Skoog in 1957. Techniques for genetic transformation include protoplast fusion, using Agrobacterium tumerfaciens and its Ti plasmids, microinjection, and electroporation. Plant tissue culture is now used for cloning plants, introducing new traits through genetic engineering, and mass producing commercial crops.
Plant tissue culture
Plant tissue culture is a technique used to propagate plants asexually under controlled laboratory conditions. It involves culturing plant cells, tissues, or organs on artificial nutrient media. The key principles are totipotency, where plant cells can regenerate into whole plants, and plasticity, where plants can alter their growth to suit their environment. There are various types of plant tissue culture including embryo, seed, meristem, cell, protoplast, callus, pollen, and organ culture. Important applications include producing virus-free plants through meristem culture and generating novel hybrids through protoplast fusion.
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Plant tissue culture involves growing plant cells, tissues, or organs in a sterile nutrient medium. Key developments included the discovery that single plant cells could be cultured in 1902, the role of growth regulators in the 1930s, and the development of techniques like protoplast fusion and genetic engineering in the 1970s-1980s. Successful tissue culture involves explant preparation, multiplication of cultures, rooting, and transplantation to the greenhouse. Various methods can be used to introduce foreign DNA, including Agrobacterium infection, microinjection, and electroporation.
application of tissue culture in crop improvement
The document discusses various applications of tissue culture and genetic engineering in crop improvement. It provides examples of transgenic plants developed with improved nutritional quality, biotic and abiotic stress tolerance, and for producing pharmaceuticals. Specifically, it describes the development of golden rice with enhanced vitamin A content and Bt crops with insect resistance. The use of genetic engineering to develop herbicide and disease resistant crops as well as plants with enhanced traits like drought and salt tolerance is also covered.
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Plant tissue culture
Biology project on plant tissue culture of brahmi leaves, and a gist of the general calyx induction.
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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.
Plant tissue culture,its methods, advantages,disadvantages and applications.
Plant tissue culture is the most widely used technique for growing very large number of plant using a very small part of the main plant(explant). Tissue culturing is very common for many popular and demanding crops.Few of them discussed here are Potato,Papaya,Pinepple,Banana,Gerbera,Sunflower,Orchids
1. propagation by plant tissue culture
Plant tissue culture techniques are used to grow plant cells, tissues, or organs in sterile conditions. This allows for cloning of plants through micropropagation, where a small piece of plant tissue is grown in a nutrient medium, allowing for mass propagation. The document discusses the history and development of tissue culture, including early experiments in the 1800s and discoveries of growth hormones like auxin and cytokinin that allowed cultured cells and tissues to proliferate. It also outlines the stages of micropropagation including establishment, multiplication, root formation, and acclimatization of plants.
Establishment Of Cell Suspension Callus Culture from Leaves Andrographis pani...
Establishment Of Cell Suspension Callus Culture from Leaves Andrographis paniculata sp. and Micropropagation of Borreria laevicaulis sp.
Prospect of tissue culture in india
1. Introduction: Tissue Culture is the in vitro culture of cells, tissues, organs or whole plant under controlled nutritional and environmental Conditions(T. Thorp, 2007).
The science of plant tissue culture takes its roots from the discovery of Cells (Robert Hooke in 1665) and 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 plants.
2. Plant Tissue Culture: Past & Present Prospects
In 1902, a German physiologist, Gottieb Haberlandt for the first time attempted to culture isolated single palisade cells from leaves in knop’s salt solution.
The cell remained alive for up to 1 month, increased in size, accumulated starch but failed to divide.
Though he was unsuccessful but he laid the foundation of tissue culture so he is regarded as Father of Plant Tissue Culture.
In the Subsequent years different landmark discoveries were made. Some of them are:
Use of specialized media for aseptic culture of Orchid seeds (Knudson, 1925) and other workers also demonstrated that plants could be propagated in vitro from the minuscule seeds of the Orchidaceae.
Further culture of other plant tissue was not possible due to lack of knowledge of the specific hormones to be added to the culture media.
This limitation was overcomed by the elucidation of the nature of Auxin, IAA, by Thimann and Went(1930) that plants would be subsequently regenerated through the use of IAA or its analogs.
Discovery of Cytokinins, specially Kinetin(6-furfurylaminopurine) by Miller et al. (1956), the regeneration of intact plants from tissue of many herbaceous species became a practical reality.
Meristem tip culture for the production of the virus free plants
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
3. plant tissuce culture media
Plant tissue culture media provides nutrients and environment for plant growth in vitro. It contains inorganic salts that provide minerals, organic compounds like sugars and vitamins, and gelling agents. Common media include Murashige and Skoog, Gamborg's B5, and Linsmaier-Skoog. These supply essential elements like nitrogen, phosphorus, and potassium as well as trace elements needed for plant development. Media is prepared by adding ingredients to water, adjusting pH, adding gelling agent, sterilizing by autoclaving, and allowing it to solidify before use in culture.
plant tissue culture by thanuja
This document provides an overview of plant tissue culture. It defines tissue culture as the in vitro cultivation of plant cells or tissues under aseptic conditions on a nutrient medium. The history and key figures in the development of plant tissue culture are discussed. Details are provided on nutrient requirements, preparation and sterilization of culture media, basic laboratory requirements, establishment of cultures from explants, and types of growth and cultures. The advantages and applications of plant tissue culture are also summarized.
History of plant biotechnology
Plant biotechnology is based on cellular totipotency and genetic transformation. Key developments include the Cell Theory of Schleiden and Schwann in the 1830s-1840s, Gottlieb Haberlandt's experiments in the late 1800s demonstrating plant cell totipotency, and Frederick Griffith's 1928 discovery of genetic transformation in bacteria. In 1983, three groups introduced the first transgenic plants by transferring bacterial genes into tobacco using Agrobacterium tumefaciens-mediated transformation. Scientists hoped this new technology could introduce important agronomic genes into crop species.
Poster presentation ptc - kamlesh
This document summarizes plant tissue culture techniques. It discusses the history of plant tissue culture from 1902 to 1987, highlighting important discoveries. The principles of totipotency and plasticity are described. Different types of plant tissue culture are listed, including embryo culture, seed culture, and meristem culture. Applications such as producing virus-free plants, mass propagation, and creating hybrids through protoplast fusion are covered.
Lab soil
This document outlines an experiment to determine the effect of temperature, humidity, and incubation time on the decomposition of organic compounds by soil microorganisms. Students will incubate soil with carbohydrates, proteins, and lipids at different temperatures and humidity levels for varying durations. They will then test for the presence of these compounds to see which are most decomposed under each condition. The experiment aims to show how environmental factors influence the activity of soil microbes and the carbon cycle of organic matter decomposition.
Plant tissue culture
Plant tissue culture involves growing plant cells, tissues, or organs in sterile conditions on a nutrient culture medium. Key aspects of plant tissue culture include selecting and sterilizing explant tissue, transferring it to a solid culture medium containing hormones, and periodically moving the growing tissue to new media for multiplication. The growing plantlets are eventually transferred to pots and grown in a greenhouse. Plant tissue culture allows for conservation of endangered species, production of virus-free plants, hybrid regeneration, and faster plant growth.
Plant tissue culture By Allah Dad Khan
This document provides a history of major developments in plant tissue culture from 1902 to 2005. Some of the key events include:
- 1902: Haberlandt proposed the concept of in vitro cell culture.
- 1954: Muir was the first to break callus tissues into single cells.
- 1955: Skoog and Miller discovered kinetin as a cell division hormone.
- 1960: Cocking was the first to isolate protoplasts by enzymatic degradation of cell walls.
- 1964: Guha and Maheshwari produced the first haploid plants from pollen grains through anther culture.
Secondary metabolities, Shikonin production
Secondary metabolites are organic compounds produced by plants and organisms that are not essential for growth or reproduction. They play important roles in defense against herbivores and pathogens. Approximately 1500 new secondary metabolite molecules are identified from plants each year, around 30% of which show some biological activity. Secondary metabolites have many applications in medicine, food, cosmetics, and other industries. Plant tissue culture is used to produce many important secondary metabolites in a controlled environment, as production from native plants can be limited by environmental and geographical factors. Common production methods include cell suspension cultures, hairy root cultures, and immobilized cell cultures. Factors like media composition, temperature, pH, and elicitors can influence metabolite yield. An example is the production
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Culture stage 1 initiation stage
Introduction
Advantages of Micropropagation over the conventional methods
History
Stages of Micropropagation
1. Stage 0; Preparative stage
2. Stage 1; Initiation of aseptic cultures
A) Explant
B) Sterilization
C) Browning of medium
Factors affecting initiation stage
Conclusions
References
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The document discusses micropropagation, which is a type of tissue culture used to rapidly produce many uniform plants from small plant parts like meristems. It involves four stages: establishing an aseptic culture, multiplying propagules in nutrient media, pre-transplant rooting, and transplanting to soil. Micropropagation has advantages like producing many clean, genetically identical plants quickly and independently of seasons. It requires specialized facilities and expertise. Potential problems include contamination, tissue browning, abnormal growth, and shock from in vitro to in vivo conditions. Micropropagation has applications for rapidly increasing stocks of superior varieties and producing disease-free plants.
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Tissue culture is the process of growing plant cells, tissues or organs in an artificial, sterile environment. It involves removing plant cells and placing them in a nutrient solution supplemented with hormones, vitamins and minerals. Key requirements include using appropriate explant tissue, a suitable growth medium, aseptic conditions, and growth regulators. The document outlines various tissue culture techniques such as micropropagation, callus culture, somatic embryogenesis, organogenesis, and anther/pollen/ovule/ovary culture. Factors affecting successful tissue culture include the plant genotype, explant source, nutrient composition of the growth medium, and control of environmental factors like light, temperature and sterility.
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This document summarizes micropropagation techniques used to rapidly multiply plants in vitro. It discusses explant selection, initiation and establishment of aseptic cultures, multiplication of plantlets, and acclimatization. Micropropagation allows for mass production of disease-free plants through small propagules in controlled environments. The document also covers plant genetic transformation methods using Agrobacterium tumefaciens to introduce exogenous DNA and produce transgenic plants with desired traits.
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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.
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The document describes a seminar presentation on using plant tissue culture techniques to induce variation and improve crops. It provides background on tissue culture, discusses types of culture and variation that can occur. It also presents a case study on developing insect-resistant transgenic brinjal (eggplant) and studies on tissue culture-induced variation in tomato and torenia plants. The conclusion states that tissue culture has been valuable for plant biotechnology and improving food security.
Hairy root culture, multiple shoot culture ,
Definition of hairy root culture ,multiple shoot culture ,Production of hairy root and multiple shoot , advantages an disadvantages of hairy root and multiple shoot culture, Sterilization and sterilizing agents wit concentration and exposure time
PLANT TISSUE CULTURE.pptx
Plant tissue culture involves growing plant cells, tissues or organs under sterile conditions on a nutrient medium. It has a history dating back to the 1930s and was pioneered by scientists like Haberland and White. Techniques include using explants from plants, sterilizing them, and culturing them on nutrient media containing salts, sugars and growth regulators. This allows for dedifferentiation of cells and regeneration of whole plants. Micropropagation specifically refers to rapidly multiplying shoots in culture. It has five stages - selection of stock plants, initiation of culture, shoot multiplication, rooting, and hardening. It has advantages like producing many pathogen-free plants quickly from small explants.
Cell culture techniques
Equipments used , types of culture and media, subculturing, secondary culture, finite & continuous cell lines, cryopreservation and applications of cell culture
report on in vitro seed production and seed immobilization
This document describes a student's project report on in vitro seed production and seed immobilization. It includes an introduction describing the importance of seed viability testing and in vitro techniques. It also describes the objectives of the project which are: 1) determining mung bean seed viability, 2) preparing MS medium stock solutions, 3) in vitro mung bean seed germination, and 4) artificial seed production using sodium alginate immobilization. The document includes sections on review of literature, materials and methods, results and discussion, and references.
4th tissue culture
This document discusses various types of plant tissue culture techniques. It defines explants as sterile pieces of plant material used to initiate cultures. Young, rapidly growing tissue is preferred for explants. Common types of in vitro culture discussed include callus culture, cell suspension culture, organ culture, and protoplast culture. Aseptic technique and sterilization methods like heating, filtration, and chemicals are used to prevent contamination during culture.
58833070-17a4-4647-ada7-8edbb27bfb1a.pdf
Callus culture is the growth of undifferentiated cells from plant explants on artificial nutrient media. The history of callus culture began in the 1920s with experiments on carrot tissue. Key developments included the discovery of the plant hormone auxin in 1928 and the establishment of callus cultures using auxin in the 1930s-1940s. Callus cultures are initiated from explants placed on media containing auxin and cytokinin, then subcultured regularly. Callus appears as an unorganized mass of cells that can be used for experiments or regenerated into plantlets.
Steps involved in micropropagation
The document describes the steps involved in micropropagation:
1. Explant selection from a donor plant
2. Establishment of the explant in culture media
3. Callus development and cell division from the explant
4. Development of plantlets from the callus tissue
5. Hardening or acclimatization of the plantlets for transplanting.
Steps involved in micropropagation
The document describes the steps involved in micropropagation:
1. Explant selection from a donor plant
2. Establishment of the explant in culture media
3. Callus development and cell division from the explant
4. Development of plantlets from the callus tissue
5. Hardening or acclimatization of the plantlets for transplanting.
Plant tissue culture
Tissue culture is a technique where cells, tissues or whole plants are grown in a sterile nutrient culture medium under controlled conditions. It allows for rapid vegetative propagation of plants. Key steps include sterilizing equipment and explants, preparing nutrient medium, subculturing to promote growth, and rooting and hardening plantlets. Tissue culture has many applications like mass multiplication of crops and plants, eliminating diseases, and genetic modification. It is used commercially for propagating crops but contamination and rooting difficulties can be issues.
Tissue culturing
Tissue culture is a process that clones plants through micropropagation. It involves culturing plant tissues in sterile conditions with specific nutrients and hormones. There are four main stages - initiation, multiplication, rooting, and acclimatization. The multiplication stage uses cytokinins to induce shoot growth from explants like leaves or stems. Rooting uses auxins to induce root formation from shoots. The process allows for mass production of genetically identical plants independent of seasons.
Suresh yadav
This document discusses production of disease-free seedlings through tissue culture techniques. It begins with an introduction to meristem culture and how it can be used to eliminate viruses from plant materials. It then provides the step-by-step protocol for conducting meristem culture, including explant selection, surface sterilization, culture establishment, and acclimatization of plantlets. Applications of meristem culture like virus elimination, micropropagation, and facilitating international exchange of germplasm are also summarized. The document lists some advantages and disadvantages of the technique.
Applications of Plant Tissue Culture
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
Introduction to basic biotechnology - part a
1. The document discusses biotechnology applications for horticultural crops including genetically modified organisms. It describes techniques like genetic engineering, tissue culture, embryo culture and haploid breeding that can be used to develop new crop varieties.
2. Examples of traits that can be improved through biotechnology include increased yield, pest and disease resistance, and improved postharvest qualities. The document lists some GM crops that have been approved or are undergoing field tests.
3. The challenges of tropical fruit export and postharvest storage are discussed. Controlling ethylene production and perception genes could help extend shelf life of fruits like mango to increase export potential. Precision genome editing techniques may allow targeting of genes with more specificity.
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report on in vitro seed production and seed immobilization
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0.1 plant tissue culture step by step guide
- 1. By – Dr. Mafatlal M. Kher Plant tissue culture step by step guide Date & Time : Wednesday, 08 September 2021 Semester : V Program : B.Sc. Biotechnology School : School of Science Subject code : BSBO502 (Unit I) 1
- 2. Plant tissue culture step by step Guide Selection of plant species Selection of mother plant for collection of explant Explant Surface sterilization Media composition for various objectives (Callus, somatic embryo, shoot etc) Genetic fidelity Acclimatization (Primary hardening, secondary hardening, field transfer) 2
- 3. Selection of Plant Species Commercial Medicinal Horticulturally important plant Agricultural Biodiversity related: Rare, IUCN Red Data List. 3
- 4. Selection of mother/stock plant of explant collection Healthy Disease free Actively growing shoots Availability of explant High Yielding If medicinal: amount of bioactive compounds/gm of fresh or dry weight Horticulture: Fruit (quality and quantity) Conservation: If plant is rare or not 4
- 5. Explant Type Explant with predetermined meristem: Shoot tip (shoot apical meristem), Node (axillary meristem). Without predetermined meristem: Leaf, internode, petiole, petal, anther, pollen grains, unfertilized or fertilized ovule. Size Explant size 1.5 to 2.5 cm length for node or minimum one axillary bud ( for nodal explant in a tree species) or with 2-3 axillary buds in case of herbaceous plants like mint, brahmi etc. Shoot tip culture < 0.2 cm Orientation Node: Upright position (natural position), Leaf natural position lower surface must be in contact with media, therefore slants are prepared for leaf or explants without predetermined meristem, sometimes such explants can cultured in petridishesh too. 5
- 6. Discussion Q: If I have inoculated 100 shoot tip and 100 nodes on shoot culture medium: Which one will perform better? And Why? Answer: Node, because it is having more vigour index (more surface area, more cell, more cytoplasm). Q: Why Shoot tip culture is still in practice? Answer: Shoot tip culture is utilized for the production of Virus free plant. What is special in shoot tip ? • Active cell division, it will not allow virus to replicate. • Vascular bundles are abs, as virus transmitted via vascular bundle • High cell metabolic activity, that will kill virus articles during cell division. • Therefore, shoot tip culture is preferred for the production of virus free plants. 6
- 7. Surface sterilization of explants Types of surface sterilizing agents and their concentrations, combinations and duration of the treatment. 7 Sterilizing agent Concentration used (%) Duration (min) Effectiveness Calcium hypochlorite 9-10 5-30 Very good Sodium hypochlorite 20% v/v commercial solution 5-30 Very good Hydrogen peroxide 10-12 5-15 Good Bromine water 1-2 2-10 Very good Silver nitrate 1 5-30 Good Mercuric chloride 0.1-1 2-10 Very good Ethyl alcohol 70 30 sec to 2 Good Isopropyl alcohol 70 30 sec to 2 Good Bavistin (50% WP carbendazim. Carbendazim 50%WP) 2% 2-10 Good (Antifungal)
- 8. Basic steps for:- Callus culture Somatic embryogenesis Axillary shoot or shoot tip culture 8
- 9. Media composition Callus culture 1. Callus induction medium (Keep cultures for one to two months) 2. Subculture on shoot induction medium (1-2 months) 3. Multiplication 4. Elongation of shoots 5. Rooting of shoots 6. Genetic fidelity test 7. Primary hardening 8. Secondary hardening 9. Field transfer 9
- 10. Media composition Somatic embryogenesis 1. Somatic embryo induction medium 2. Maturation of somatic embryo 3. Somatic embryo germination medium 4. Genetic fidelity test 5. Primary hardening 6. Secondary hardening 7. Field transfer 10
- 11. Media composition Axillary shoot/Shoot tip culture 1. Shoot induction 2. Shoot multiplication 3. Elongation of shoots 4. Rooting of shoots 5. Genetic fidelity test 6. Primary hardening 7. Secondary hardening 8. Field transfer 11