The document describes the process of somatic embryogenesis. It involves 7 key steps:
1) Induction of embryogenesis from explant tissue on media supplemented with auxin
2) Development of somatic embryos through globular, heart, and torpedo stages of growth
3) Maturation of embryos with the formation of root and shoot meristems and cotyledons
4) Conversion of mature embryos to plantlets through germination on auxin-free media
Factors like explant type, growth regulators, and genotype influence the process. Somatic embryos differ from zygotic embryos in lacking a seed coat and having greater potential for propagation but weaker plantlets.
This document discusses anther and pollen culture techniques. It provides a brief history of the development of these techniques from the 1950s onward. It then describes the process of anther culture, where anthers are cultured in nutrient medium to produce haploid callus or embryos. Pollen or microspore culture involves isolating pollen grains from anthers and culturing them. The goal is to produce haploid embryos or callus that can develop into haploid plantlets. Key factors that influence success include the genotype, microspore stage, culture medium, temperature, and physiological status of the donor plant. Anther culture has applications in mutation studies, plant breeding, and secondary metabolite production.
This document discusses meristem culture and shoot tip culture techniques. It describes the three stages of meristem culture: establishment, multiplication, and root regeneration. Shoot tips less than 1 mm are excised and cultured on medium supplemented with hormones like cytokinins and auxins to promote growth. Meristem culture allows for virus elimination, micropropagation, genetic resource preservation, and facilitates international plant exchange. It is an effective method for producing disease-free plants.
1. Acclimatization is the process by which plants adapt to changes in their environment over multiple generations through natural selection.
2. It requires genetic variability in introduced plant materials and occurs more readily in cross-pollinated species and annual crops.
3. Examples of acclimatization include humans developing more red blood cells at high altitudes and plants surviving freezing temperatures if the temperature drops gradually over time rather than suddenly.
Synthetic seeds are encapsulated somatic embryos or shoot buds that can be used for planting like traditional seeds. They allow for clonal propagation of plants that are difficult to reproduce through traditional seeds, including some fruit crops. The production of synthetic seeds involves inducing somatic embryogenesis in callus cultures, maturing the embryos, and encapsulating them in a protective gel before planting. This allows genetic material to be stored and dispersed while avoiding issues with seed-borne diseases, low seed viability, and difficulties reproducing species that lack traditional seeds.
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.
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.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
This document discusses anther and pollen culture techniques. It provides a brief history of the development of these techniques from the 1950s onward. It then describes the process of anther culture, where anthers are cultured in nutrient medium to produce haploid callus or embryos. Pollen or microspore culture involves isolating pollen grains from anthers and culturing them. The goal is to produce haploid embryos or callus that can develop into haploid plantlets. Key factors that influence success include the genotype, microspore stage, culture medium, temperature, and physiological status of the donor plant. Anther culture has applications in mutation studies, plant breeding, and secondary metabolite production.
This document discusses meristem culture and shoot tip culture techniques. It describes the three stages of meristem culture: establishment, multiplication, and root regeneration. Shoot tips less than 1 mm are excised and cultured on medium supplemented with hormones like cytokinins and auxins to promote growth. Meristem culture allows for virus elimination, micropropagation, genetic resource preservation, and facilitates international plant exchange. It is an effective method for producing disease-free plants.
1. Acclimatization is the process by which plants adapt to changes in their environment over multiple generations through natural selection.
2. It requires genetic variability in introduced plant materials and occurs more readily in cross-pollinated species and annual crops.
3. Examples of acclimatization include humans developing more red blood cells at high altitudes and plants surviving freezing temperatures if the temperature drops gradually over time rather than suddenly.
Synthetic seeds are encapsulated somatic embryos or shoot buds that can be used for planting like traditional seeds. They allow for clonal propagation of plants that are difficult to reproduce through traditional seeds, including some fruit crops. The production of synthetic seeds involves inducing somatic embryogenesis in callus cultures, maturing the embryos, and encapsulating them in a protective gel before planting. This allows genetic material to be stored and dispersed while avoiding issues with seed-borne diseases, low seed viability, and difficulties reproducing species that lack traditional seeds.
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.
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.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
This document discusses distant hybridization and various techniques used to produce haploid plants. Distant hybridization refers to crosses between individuals of different plant species or genera. Such crosses can result in fully fertile, partially fertile, or fully sterile offspring depending on chromosomal homology. Androgenesis and gynogenesis are techniques used to induce haploid plants from male and female gametes, respectively. Androgenesis involves culturing immature anthers or isolated microspores while gynogenesis involves culturing unpollinated flower parts. Wide hybridization is also used to induce maternal haploids. Factors like genotype, developmental stage, and culture conditions influence haploid induction and regeneration.
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.
- 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.
Somatic embryogenesis, in plant tissue culture 2KAUSHAL SAHU
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
Embryo culture involves growing plant embryos artificially in order to enhance survival rates. It is commonly used to rescue weak or immature embryos that may not otherwise survive to become viable plants. The process involves excising embryos from seeds or ovaries and placing them onto sterile nutrient-rich media under suitable temperature, light, and humidity conditions. Embryo culture has various applications in plant breeding, including shortening breeding cycles, overcoming seed dormancy, producing hybrids, and conserving plant germplasm. It is an important technique in modern plant breeding and development of new crop varieties.
Anther culture is a technique where anthers are excised from flower buds and cultured to produce haploid plants. The first report of haploid tissue from anther culture was in 1964-1966 in Datura pollen grains. Over 250 species have been produced through anther culture, most commonly in families like Solanaceae, Cruciferae, and Poaceae. Haploid plants are useful for identifying recessive traits, eliminating lethal genes, and producing homozygous diploid plants more quickly. There are several pathways that microspores can follow during anther culture, such as symmetric or asymmetric division, to produce haploid plants. Successful anther culture requires optimizing various factors like donor plant genotype, anther
Cell suspension culture involves growing single plant cells or small cell aggregates in agitated liquid medium. It allows studying cellular events during growth and development without the limitations of callus culture. An ideal suspension culture consists of only uniformly growing single cells. It is established by transferring friable callus pieces to agitated medium, then filtering and subculturing the dispersed cells. Suspension culture offers insights into cell physiology and is useful for cloning, secondary metabolite production, and mutagenesis studies. While it addresses issues with callus culture, cell suspension cultures can have decreasing productivity over time and slow growth.
Somatic hybridization is a technique used to create hybrid plants by fusing isolated plant cells called protoplasts from two different plant species or varieties. This fusion occurs under in vitro conditions and can result in symmetric hybrids that contain chromosomes from both parents, or asymmetric hybrids that lose chromosomes from one parent. Cybrids are a type of hybrid where the nucleus comes from one species but the cytoplasm, including chloroplasts and mitochondria, comes from both parental species. Somatic hybridization and cybrid production allow for novel combinations of genes that can provide agricultural benefits like stress resistance but technical challenges remain in regenerating hybrid plants.
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.
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
This document discusses somaclonal variation, which refers to genetic variation that arises during tissue culture or plant regeneration from cell cultures. It provides definitions and history of the term as coined by Larkin and Scowcroft in 1981. The document outlines the various causes and types of somaclonal variation including physiological, genetic, and biochemical causes. It also describes methods for generating somaclonal variation both with and without in vitro selection. Finally, it discusses applications for detecting and isolating somaclonal variants, particularly for developing disease resistance in various crop species.
Genetic variations can occur in plants produced through plant tissue culture and be detected as changes in genetic characteristics or phenotypes. Variations commonly include changes in chromosome number and structure. Regenerated plants with chromosomal changes often show alterations in traits like leaf shape and color, growth rate, and fertility. These heritable mutations can persist when plants are transplanted to fields. Somaclonal variations are caused by genetic factors like pre-existing variations in explant cells or mutations during tissue culture, and can result in changes in plant characteristics that are useful for crop improvement.
1) Germplasm conservation involves preserving genetic material, such as seeds, cells, tissues, and body parts, through in-situ and ex-situ methods to maintain biodiversity and provide resources for breeding programs.
2) Cryopreservation at ultra-low temperatures in liquid nitrogen is an important ex-situ technique that can preserve germplasm long-term without subculturing. It involves preculturing plant materials, treating with cryoprotectants, and either slow-freezing or vitrification prior to storage in liquid nitrogen.
3) A case study demonstrates the successful cryopreservation of mint shoot tips using encapsulation-dehydration and PVS2-vitrification, with
This document discusses somatic embryogenesis and its consequences in cereals. It begins with an introduction to somatic embryogenesis, noting that it is a process where embryos are derived from somatic cells rather than gametes. It then covers factors that affect somatic embryogenesis like the explant source, plant growth regulators, and genotype. It also describes the stages of somatic embryogenesis and different types. The document discusses the role of somatic embryogenesis in improving cereals through somaclonal variation and disease resistance. It concludes that somatic embryogenesis is a model for plant breeding and genetic improvement.
A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Seed dormancy allows seeds to remain dormant during unfavorable conditions until conditions become suitable for germination. There are two main types of dormancy - primary and secondary. Primary dormancy occurs due to internal factors like hormones, while secondary dormancy is caused by external factors like temperature. Dormancy can be overcome through methods like scarification, stratification, hormone treatment, and photoperiod manipulation. Seed dormancy provides important biological benefits like survival during drought or frost and dispersal to new areas.
Organogenesis, in plant tissue cultureKAUSHAL SAHU
Introduction
Definition
Types of organogenesis
Organogenesis through callus formation (indirect organogenesis)
Growth regulators for indirect organogenesis
Organogenesis through adventitious organ (direct organogenesis)
Growth regulators for direct organogenesis
Factor affecting the soot bud differentiation
Organogenic differentiation
Application of organogenesis
Conclusion
References
Anther culture is an in vitro technique used to produce haploid plants from male gamete cells. Haploid cells contain only one set of chromosomes. The first report of anther culture was in the 1970s, while the first natural occurrence of haploids was observed in 1922 in Datura plants. Anthers containing microspores are cultured on nutrient media supplemented with hormones and sugars. Depending on the species, haploid plants or callus can regenerate from the cultured anthers within 3-8 weeks. Haploid plants are useful for breeding programs as they can be doubled to generate fertile, homozygous plants in one generation.
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.
Micropropagation, or in vitro clonal propagation, allows for the rapid multiplication of plant materials to produce many progeny plants using modern tissue culture methods. Clonal propagation is the asexual reproduction of genetically identical individuals. Micropropagation involves selecting plant material, sterilizing explants, culturing them on growth media, and multiplying shoots through successive transfers. Shoots are then rooted and acclimatized. The process uses controlled conditions and growth regulators to stimulate organogenesis or somatic embryogenesis. Micropropagation has applications for commercial scale propagation of new varieties, eliminating diseases, and cloning plants that are difficult to propagate conventionally.
This document discusses distant hybridization and various techniques used to produce haploid plants. Distant hybridization refers to crosses between individuals of different plant species or genera. Such crosses can result in fully fertile, partially fertile, or fully sterile offspring depending on chromosomal homology. Androgenesis and gynogenesis are techniques used to induce haploid plants from male and female gametes, respectively. Androgenesis involves culturing immature anthers or isolated microspores while gynogenesis involves culturing unpollinated flower parts. Wide hybridization is also used to induce maternal haploids. Factors like genotype, developmental stage, and culture conditions influence haploid induction and regeneration.
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.
- 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.
Somatic embryogenesis, in plant tissue culture 2KAUSHAL SAHU
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
Embryo culture involves growing plant embryos artificially in order to enhance survival rates. It is commonly used to rescue weak or immature embryos that may not otherwise survive to become viable plants. The process involves excising embryos from seeds or ovaries and placing them onto sterile nutrient-rich media under suitable temperature, light, and humidity conditions. Embryo culture has various applications in plant breeding, including shortening breeding cycles, overcoming seed dormancy, producing hybrids, and conserving plant germplasm. It is an important technique in modern plant breeding and development of new crop varieties.
Anther culture is a technique where anthers are excised from flower buds and cultured to produce haploid plants. The first report of haploid tissue from anther culture was in 1964-1966 in Datura pollen grains. Over 250 species have been produced through anther culture, most commonly in families like Solanaceae, Cruciferae, and Poaceae. Haploid plants are useful for identifying recessive traits, eliminating lethal genes, and producing homozygous diploid plants more quickly. There are several pathways that microspores can follow during anther culture, such as symmetric or asymmetric division, to produce haploid plants. Successful anther culture requires optimizing various factors like donor plant genotype, anther
Cell suspension culture involves growing single plant cells or small cell aggregates in agitated liquid medium. It allows studying cellular events during growth and development without the limitations of callus culture. An ideal suspension culture consists of only uniformly growing single cells. It is established by transferring friable callus pieces to agitated medium, then filtering and subculturing the dispersed cells. Suspension culture offers insights into cell physiology and is useful for cloning, secondary metabolite production, and mutagenesis studies. While it addresses issues with callus culture, cell suspension cultures can have decreasing productivity over time and slow growth.
Somatic hybridization is a technique used to create hybrid plants by fusing isolated plant cells called protoplasts from two different plant species or varieties. This fusion occurs under in vitro conditions and can result in symmetric hybrids that contain chromosomes from both parents, or asymmetric hybrids that lose chromosomes from one parent. Cybrids are a type of hybrid where the nucleus comes from one species but the cytoplasm, including chloroplasts and mitochondria, comes from both parental species. Somatic hybridization and cybrid production allow for novel combinations of genes that can provide agricultural benefits like stress resistance but technical challenges remain in regenerating hybrid plants.
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.
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
This document discusses somaclonal variation, which refers to genetic variation that arises during tissue culture or plant regeneration from cell cultures. It provides definitions and history of the term as coined by Larkin and Scowcroft in 1981. The document outlines the various causes and types of somaclonal variation including physiological, genetic, and biochemical causes. It also describes methods for generating somaclonal variation both with and without in vitro selection. Finally, it discusses applications for detecting and isolating somaclonal variants, particularly for developing disease resistance in various crop species.
Genetic variations can occur in plants produced through plant tissue culture and be detected as changes in genetic characteristics or phenotypes. Variations commonly include changes in chromosome number and structure. Regenerated plants with chromosomal changes often show alterations in traits like leaf shape and color, growth rate, and fertility. These heritable mutations can persist when plants are transplanted to fields. Somaclonal variations are caused by genetic factors like pre-existing variations in explant cells or mutations during tissue culture, and can result in changes in plant characteristics that are useful for crop improvement.
1) Germplasm conservation involves preserving genetic material, such as seeds, cells, tissues, and body parts, through in-situ and ex-situ methods to maintain biodiversity and provide resources for breeding programs.
2) Cryopreservation at ultra-low temperatures in liquid nitrogen is an important ex-situ technique that can preserve germplasm long-term without subculturing. It involves preculturing plant materials, treating with cryoprotectants, and either slow-freezing or vitrification prior to storage in liquid nitrogen.
3) A case study demonstrates the successful cryopreservation of mint shoot tips using encapsulation-dehydration and PVS2-vitrification, with
This document discusses somatic embryogenesis and its consequences in cereals. It begins with an introduction to somatic embryogenesis, noting that it is a process where embryos are derived from somatic cells rather than gametes. It then covers factors that affect somatic embryogenesis like the explant source, plant growth regulators, and genotype. It also describes the stages of somatic embryogenesis and different types. The document discusses the role of somatic embryogenesis in improving cereals through somaclonal variation and disease resistance. It concludes that somatic embryogenesis is a model for plant breeding and genetic improvement.
A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Seed dormancy allows seeds to remain dormant during unfavorable conditions until conditions become suitable for germination. There are two main types of dormancy - primary and secondary. Primary dormancy occurs due to internal factors like hormones, while secondary dormancy is caused by external factors like temperature. Dormancy can be overcome through methods like scarification, stratification, hormone treatment, and photoperiod manipulation. Seed dormancy provides important biological benefits like survival during drought or frost and dispersal to new areas.
Organogenesis, in plant tissue cultureKAUSHAL SAHU
Introduction
Definition
Types of organogenesis
Organogenesis through callus formation (indirect organogenesis)
Growth regulators for indirect organogenesis
Organogenesis through adventitious organ (direct organogenesis)
Growth regulators for direct organogenesis
Factor affecting the soot bud differentiation
Organogenic differentiation
Application of organogenesis
Conclusion
References
Anther culture is an in vitro technique used to produce haploid plants from male gamete cells. Haploid cells contain only one set of chromosomes. The first report of anther culture was in the 1970s, while the first natural occurrence of haploids was observed in 1922 in Datura plants. Anthers containing microspores are cultured on nutrient media supplemented with hormones and sugars. Depending on the species, haploid plants or callus can regenerate from the cultured anthers within 3-8 weeks. Haploid plants are useful for breeding programs as they can be doubled to generate fertile, homozygous plants in one generation.
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.
Micropropagation, or in vitro clonal propagation, allows for the rapid multiplication of plant materials to produce many progeny plants using modern tissue culture methods. Clonal propagation is the asexual reproduction of genetically identical individuals. Micropropagation involves selecting plant material, sterilizing explants, culturing them on growth media, and multiplying shoots through successive transfers. Shoots are then rooted and acclimatized. The process uses controlled conditions and growth regulators to stimulate organogenesis or somatic embryogenesis. Micropropagation has applications for commercial scale propagation of new varieties, eliminating diseases, and cloning plants that are difficult to propagate conventionally.
Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on nutrient media. Key factors include the growth media, environmental conditions, and the explant source tissue. Plant cells have the abilities of totipotency, dedifferentiation, and competency. Tissue culture is used for micropropagation, genetic modification, pathogen elimination, and more. Common types include callus, organ, and embryo culture. Somatic embryogenesis and organogenesis allow regeneration of whole plants. Tissue culture can generate somaclonal variation through genetic or epigenetic changes.
1. Zygotic embryogenesis is the process by which a zygote undergoes differentiation into a mature embryo through cell division and growth.
2. Types of embryos include zygotic embryos formed by fertilization and non-zygotic embryos like somatic and parthenogenic embryos.
3. In vitro embryogenesis techniques like embryo culture can rescue hybrid embryos from wide crosses that experience post-fertilization abortion, allowing the production of rare hybrids. Embryo culture involves excising embryos and providing a nutrient medium for growth.
Somatic embryogenesis and artificial seed productionArvind Yadav
This document discusses somatic embryogenesis and artificial seed production. It describes the two main types of somatic embryogenesis (indirect and direct), the steps involved in the process, and factors that affect it such as genotype, explant type, growth regulators, and nitrogen source. It also covers embryo maturation, secondary somatic embryogenesis, synchronization of embryo development, and production of artificial or synthetic seeds by encapsulating somatic embryos. The goal is large-scale clonal propagation of plants through synthetic seed technology.
Clonal propagation invitro is called micropropagation.In micropropagation, apical meristem is cultivated. so this technique is also known as meristem culture or mericlones.Webber used the word ‘clone’for first time to apply for cultivated plants that were propagated vegetatively.It indicates that plants grown from such vegetative parts are not individuals in the ordinary sense, but are simply transplanted parts of the same individual and such plants are identical.
This document summarizes various plant tissue culture techniques including embryogenesis, organogenesis, micropropagation, callus culture, cell suspension culture, anther culture, shoot tip culture, protoplast culture, axillary bud culture, and hairy root culture. It describes the applications of each technique such as clonal propagation, production of haploids, secondary metabolite production, virus elimination, and genetic conservation. Direct and indirect organogenesis are also summarized.
This document discusses in vitro plant breeding techniques. It describes how plant cells, tissues, and organs can be cultured under controlled conditions in glass or plastic vessels with defined growth media. Plant cells have three key abilities - totipotency, dedifferentiation, and competency - that allow regeneration of whole plants in tissue culture. The ratio of auxin and cytokinin plant hormones can determine whether roots or shoots develop. Somatic embryogenesis is described as the formation of embryo-like structures from somatic cells that can develop into whole plants similarly to zygotic embryos.
1. Somatic embryogenesis is a process where embryos are formed from somatic plant cells, such as leaf cells, that are not normally involved in embryo formation. 2. These somatic embryos develop through similar stages as zygotic embryos, including globular, heart-shaped, torpedo-shaped, and cotyledonary stages. 3. Somatic embryogenesis can occur through direct embryogenesis from explant tissue or indirect embryogenesis which involves first forming a callus that then develops embryos.
This document discusses in vitro plant breeding techniques. It describes in vitro culture as the growth and development of plant tissues, cells, or protoplasts under controlled artificial conditions using glass or plastic vessels. The key abilities of plants that allow for in vitro culture are totipotency, dedifferentiation, and competency. Important factors for successful in vitro culture include growth media, environmental conditions, and the explant source. The document outlines various types of in vitro culture including seed, embryo, organ, callus, cell suspension, and protoplast cultures. It also describes plant regeneration pathways such as microcutting, organogenesis, and somatic embryogenesis.
micropropagation- a very useful technology in plant tissue culture.YoGeshSharma834784
Plant tissue culture, also known as micropropagation, involves culturing plant cells, tissues or organs on nutrient media under sterile conditions. The ratio of two key hormones, auxin and cytokinin, determines whether roots or shoots develop. Micropropagation is used for clonal propagation of plants, germplasm preservation, studying somaclonal variation, embryo culture, haploid production, and in vitro hybridization. It allows for the rapid, mass production of genetically identical plantlets while providing benefits such as being disease-free.
This document discusses plant tissue culture and micropropagation. It describes how plant cells, tissues or organs can be cultured in vitro on nutrient media to rapidly multiply stock plant materials. Two key plant hormones, auxin and cytokinin, influence shoot versus root development. Micropropagation is used for clonal propagation, germplasm preservation, and producing disease-free plants. The process involves selection of explants, establishment of sterile culture, multiplication of shoots, rooting, and acclimatization. Organogenesis and somatic embryogenesis are two regeneration pathways used.
Plant tissue culture techniques allow for the growth and development of plant cells, tissues, organs, or protoplasts in sterile conditions on nutrient media. There are several types of in vitro culture including callus culture, organ culture, and somatic embryogenesis. Plant cells have the properties of totipotency, dedifferentiation, and competency which allow regeneration of whole plants from single cells. Tissue culture is important for applications such as crop improvement, mass propagation, and germplasm conservation.
This document provides information about a seminar on micropropagation techniques in fruit crops. It discusses the need for micropropagation due to issues like seasonal limitations and virus transmission. The advantages of micropropagation include producing true-to-type plants, overcoming seasonal constraints, and allowing large-scale multiplication. The document outlines the stages of micropropagation including establishment, proliferation, rooting, and acclimatization. It also describes different approaches like axillary budding and somatic embryogenesis. Several case studies demonstrate the use of micropropagation in plants like date palm, mango, and lemon.
This document summarizes research on developing an in vitro regeneration system for indirect somatic embryogenesis in cereal crops. It begins with definitions of somatic embryogenesis and describes the key stages and factors involved. It then presents a case study on developing an indirect somatic embryogenesis system for rice. The study explores different media formulations and plant growth regulator concentrations to induce callus formation from rice scutellar explants and regenerate plantlets. Optimal conditions were identified as 3.5-5 mg/L 2,4-D for callus induction and 3 mg/L BAP with 4 mg/L NAA for shoot regeneration.
Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on nutrient media. Some key points:
- Callus culture involves growing an unorganized cell mass (callus) from explants on auxin-cytokinin media, which can then be used for micropropagation.
- Cell suspension culture breaks up callus into single cells that grow in liquid media, allowing for easier scaling up than callus.
- Micropropagation is the process of rapidly multiplying plant materials like shoots, roots or embryos in culture to produce many clonal copies of plant materials like orchids or strawberries.
- Protoplast isolation and culture allows the transfer of genes directly into plant cells without the
Somatic embryogenesis is the artificial process where an embryo develops from a somatic cell rather than a gamete. It does not involve fertilization or the formation of endosperm or a seed coat. The first observations of somatic embryogenesis were in carrot cells in 1958. Now it has been achieved in over 300 plant species. It is a valuable technique for rapid large scale propagation of plants, including for commercial and conservation purposes. The process involves four key steps - induction, maintenance, development and regeneration.
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.
Similar to Morphogenesis, organogenesis, embryogenesis & other techniques (20)
Somaclonal variation refers to genetic variation that arises in plants regenerated from tissue culture. This genetic variation can arise from changes in ploidy level, chromosome structure, or point mutations during tissue culture regeneration. Somaclonal variation provides opportunities for plant breeders to generate novel variants that may have desirable traits for crop improvement.
This document discusses protoplasm fusion and somatic hybridization, techniques covered in the course BIOTECHNOLOGY OF HORTICULTURAL CROPs. Protoplasm fusion allows the merging of plant cells from different species, while somatic hybridization uses protoplasm fusion to create hybrid plants by combining the genomes of two different plant species.
Protoplast isolation and culture allows plant cells to be studied individually. Enzymes are used to remove plant cell walls to isolate protoplasts, which can then be cultured. This lecture discusses techniques for isolating protoplasts from plant tissues and establishing cultures to regenerate new plants.
This document outlines the steps involved in plant tissue culture, which are: 1) media preparation, 2) explant selection, 3) establishment of explant in media, 4) callus development, 5) plantlet development, 6) hardening or acclimatization, and 7) open field planting. It then defines euploidy and aneuploidy, explaining that euploidy involves the entire set of chromosomes and can result in single, double, or multiple sets, while aneuploidy is having one or a few extra or missing chromosomes.
To develop a project for establishment of commercial tissue culture laboratoryHORTIPEDIA INDIA
The document provides guidelines for establishing a commercial tissue culture laboratory, including:
- The laboratory should have separate rooms for media preparation, glassware washing, sterilization, aseptic transfer, and primary culture growth to maintain cleanliness and prevent contamination.
- Facilities are needed for washing and sterilizing glassware, preparing and storing media, conducting aseptic procedures, and maintaining cultures under controlled conditions.
- Proper location, ventilation, lighting, temperature and humidity control are important considerations for laboratory design.
The document discusses developing a model cropping system by examining existing cropping systems in different regions of India, including Punjab, Himachal Pradesh and Jammu & Kashmir, the North East, Gangetic plain, Central India, Western India, humid tropics, and Telangana, Andhra Pradesh, and Karnataka. It covers agro-forestry practices and integrating forestry and agriculture farming. The goal is to gain an understanding of cropping systems currently used to inform the development of a new model.
This document discusses protoplasm fusion and somatic hybridization, which are techniques used in biotechnology for horticultural crops. Protoplasm fusion involves fusing plant cells together to create a hybrid cell, while somatic hybridization uses protoplasm fusion to combine whole plant genomes to produce a hybrid plant. These techniques can be used to introduce beneficial traits from one plant into another.
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.
This document outlines the steps involved in plant tissue culture, which are: 1) media preparation, 2) explant selection, 3) establishment of explant in media, 4) callus development, 5) plantlet development, 6) hardening or acclimatization, and 7) open field planting. It then defines euploidy and aneuploidy, explaining that euploidy involves the entire set of chromosomes and can result in single, double, or multiple sets, while aneuploidy is having one or a few extra or missing chromosomes.
This 3-credit course focuses on developing protocols for mass multiplication of horticultural crops through suspension culture techniques. Students will conduct a practical to establish suspension cultures from different explants and optimize conditions for maximum proliferation. The goal is to learn methods for large-scale commercial production of horticultural plants using biotechnological approaches.
Pruning and training techniques are important for fruit crops. Pruning is done in younger plants to establish a strong framework and in mature plants to control size and maintain production. The appropriate time for pruning depends on the specific fruit crop. For some crops like sapota and jamun, no pruning is required.
This document describes the process of nucellus culture for producing disease-free plantlets of citrus species. It involves excising nucellar tissue from unfertilized ovules and culturing it on media containing supplements like malt extract and adenine to induce embryoid formation. The embryoids are then excised and cultured on media containing gibberellic acid to develop into plantlets. Nucellus culture is useful for eliminating viral diseases from citrus and producing polyembryonic seedlings. However, it is a difficult technique requiring precise excision and culturing steps.
This document discusses various fruit cropping systems. It defines a cropping system as the crops, sequences, and management techniques used over years on a field. Sole cropping grows one crop over a large area for several years, while mixed farming combines crops and livestock. Sequential cropping plants successive crops after harvest, and ratooning allows stubble to resprout for another crop. Intercropping and relay planting involve growing two crops together through the season. Agroforestry integrates forestry and agriculture on the same land.
This document outlines the steps for in vitro plant regeneration: 1) preparing media, 2) selecting explant tissue, 3) establishing explants in media, 4) developing callus tissue, 5) developing plantlets, 6) hardening plants, and 7) planting in open fields. It also discusses using immature inflorescence, scutellar tissue from immature seeds, epidermis, and procambial tissue as explants for producing somatic embryos in plants like common wheat.
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.
To study plant tissue culture laboratory design and set upHORTIPEDIA INDIA
This document provides guidelines for setting up a plant tissue culture laboratory, including necessary equipment and design considerations. It discusses key areas of the lab such as the culture room, media preparation area, and glassware washing area. Maintaining aseptic conditions is a primary focus of the design. Proper airflow, traffic flow, and separation of clean and dirty areas are emphasized. Necessary equipment includes a laminar hood, autoclave, incubator shaker, and supplies for tissue culture work. Adherence to safety protocols is also covered.
Surface sterilization and sealing of cultures (surface sterilization of culturesHORTIPEDIA INDIA
This document provides instructions for surface sterilization of plant cultures. It discusses:
1) Explants must be sterilized to prevent microbial contamination that can kill cultured tissues.
2) Surface sterilization methods include rinsing explants, soaking in detergent and disinfectants like ethanol or mercuric chloride, then rinsing in sterile water.
3) Non-living materials like media and glassware are sterilized by autoclaving, while filters sterilize thermolabile compounds. Proper sterilization is essential to increase success of tissue culture experiments.
The document discusses explants used for clonal propagation in biotechnology of horticultural crops. It defines an explant as a tissue taken from a mother plant and cultured under aseptic conditions on a defined medium. The choice of explant depends on the type of culture to be initiated, its purpose, and the plant species. Common explant sources include shoots, leaves, stems, and roots. Explants must be surface sterilized to remove contaminants and allow contamination-free culture initiation. Sub-culturing is needed when cells reach the stationary state of growth.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
Travis Hills' Endeavors in Minnesota: Fostering Environmental and Economic Pr...Travis Hills MN
Travis Hills of Minnesota developed a method to convert waste into high-value dry fertilizer, significantly enriching soil quality. By providing farmers with a valuable resource derived from waste, Travis Hills helps enhance farm profitability while promoting environmental stewardship. Travis Hills' sustainable practices lead to cost savings and increased revenue for farmers by improving resource efficiency and reducing waste.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
Phenomics assisted breeding in crop improvementIshaGoswami9
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...Leonel Morgado
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
Immersive Learning That Works: Research Grounding and Paths ForwardLeonel Morgado
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...AbdullaAlAsif1
The pygmy halfbeak Dermogenys colletei, is known for its viviparous nature, this presents an intriguing case of relatively low fecundity, raising questions about potential compensatory reproductive strategies employed by this species. Our study delves into the examination of fecundity and the Gonadosomatic Index (GSI) in the Pygmy Halfbeak, D. colletei (Meisner, 2001), an intriguing viviparous fish indigenous to Sarawak, Borneo. We hypothesize that the Pygmy halfbeak, D. colletei, may exhibit unique reproductive adaptations to offset its low fecundity, thus enhancing its survival and fitness. To address this, we conducted a comprehensive study utilizing 28 mature female specimens of D. colletei, carefully measuring fecundity and GSI to shed light on the reproductive adaptations of this species. Our findings reveal that D. colletei indeed exhibits low fecundity, with a mean of 16.76 ± 2.01, and a mean GSI of 12.83 ± 1.27, providing crucial insights into the reproductive mechanisms at play in this species. These results underscore the existence of unique reproductive strategies in D. colletei, enabling its adaptation and persistence in Borneo's diverse aquatic ecosystems, and call for further ecological research to elucidate these mechanisms. This study lends to a better understanding of viviparous fish in Borneo and contributes to the broader field of aquatic ecology, enhancing our knowledge of species adaptations to unique ecological challenges.
Or: Beyond linear.
Abstract: Equivariant neural networks are neural networks that incorporate symmetries. The nonlinear activation functions in these networks result in interesting nonlinear equivariant maps between simple representations, and motivate the key player of this talk: piecewise linear representation theory.
Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
Current Ms word generated power point presentation covers major details about the micronuclei test. It's significance and assays to conduct it. It is used to detect the micronuclei formation inside the cells of nearly every multicellular organism. It's formation takes place during chromosomal sepration at metaphase.
Morphogenesis, organogenesis, embryogenesis & other techniques
1. 1. Media preparation
2. Explant selection
3. Establishment of explant in media
4. Callus development
5. Plantlet development
6. Hardening or acclimatization
7. Open field planting
Lecture 8: Morphogenesis, organogenesis,
embryogenesis & other techniques
Course Code : HRT 552
Course Title : BIOTECHNOLOGY OF
HORTICULTURAL CROPs
2. Introduction to Tissue Culture
⚫Tissue Culture (also known as Micropropagation or In vitro culture) is:
⚫The growing of plant cells, tissues, organs, seeds or other plant parts in
a sterile environment on a nutrient medium.
3. Steps of Micropropagation
⚫ Stage 0 – Selection & preparation of the mother plant
sterilization of the plant tissue takes place
⚫ Stage I - Initiation of culture
explant placed into growth media
⚫ Stage II - Multiplication
explant transferred to shoot media; shoots can be constantly divided
⚫ Stage III - Rooting
explant transferred to root media
⚫ Stage IV - Transfer to soil
explant returned to soil; hardened off
11. 3/27/2015 Deptt of Plant Biotechnology 11
EMBRYOGENESIS
:-
Plant embryogenesis refers to the process of development
of plant embryos, being either a sexual or asexual
reproductive process that forms new plants.
Embryogenesis may occur naturally in the plant as a result
of sexual fertilization, and those embryo are called zygotic
embryos and develop into seeds, which can germinate and
give rise to seedlings.
Plant cells can also be induced to form embryos in plant
tissue culture; these embryo are called somatic embryos.
14. Zygotic Embryogenesis:-
The zygotic embryo is formed following double
fertilization of the ovule, forming the plant
embryo and the endosperm which together go
into the seed, this process is known as zygotic
embryogenesis.
Seeds may also develop without fertilization
through pathways referred to as apomixis.
15. Somatic Embryogenesis:-
Somatic embryogenesis is a process by which
somatic cells or tissues develops into
differentiated embryos.
Embryos regenerate from somatic cells or
tissues ( haploid or diploid etc) it is termed as
Somatic Embryogenesis.
16. Somatic embryogenesis was first induced in suspension culture
(Stewart et al, 1958) and in callus culture (Reinert, 1959) of carrot,
Umbelliferae and Solanaceae dicotyledonous families have
produced somatic embryos.
SE occur most frequently in tissue culture as an alternative
organogenesis for regeneration of whole plant.
In literature, somatic embryos are referred to by many names such
as embryo like structures, adventitious or vegetative embryos,
Embryoids; and the process is termed as adventitious , asexual or
somatic embryogenesis.
19. Contd:-
INDUCTION
Development and
Maturation
Globular
Heart stage
Torpedo
Germination and
Conversion
• Globular stage: Embryo
is small and round
(multicellular).
• Heart stage (Bilateral
symmetry): Shape changes
to heart shape with more
cotyledon development.
• Torpedo shaped stage:
Consists of initial cells for
the shoot/root meristem.
• Mature stage: Embryo
becomes cylindrical.
20. Induction
Auxin required for induction
Pro embryonic masses are formed.
2,4-D are mostly used.
NAA, DICAMBA are also used.
21. Development
• Auxin must be removed for embryo development.
• Continuous use of Auxin inhibits embryogenesis.
• Stages are similar to those of Somatic embryogenesis:-
Globular
Heart
Torpedo
Cotyledonary
Germination (Conversion)
22. Maturatio
n
Require complete maturation with apical meristem,
radicle and cotyledons.
Often obtained repetitive embryony.
Storage protein production necessary.
Often require ABA for complete maturation.
ABA often required for normal morphology.
24. Routes of Somatic Embryogenesis:-
Two routes to somatic embryogenesis
Direct somatic embryogenesis
The embryos initiate directly from explants in the
absence of callus formation. Embryos are formed
due to PEDCs cell.
Indirect somatic embryogenesis
Callus from explants takes place from which
embryos are developed. Embryos are formed due to
IEDCs cells.
25. Examples of Direct Somatic Embryogenesis:-
Figure :- Isolation of mature embryo from imbibed cereal grain. (a) A curved-tip scalpel
blade is inserted beneath the Coleoptilar region of the Mature embryo; (b) With a swift and
smooth scooping motion the mature embryo is dislodged from its attachment to the
scutellum; (c) Isolated mature embryo which will be inoculated with abaxial surface in
contact with culture medium. Ganeshan et al., 2006.
26. Contd:
- Mature embryos culture in the Murashige and Skoog, 1962
medium with supplements 1gm/l enzymatic casein
hydrolysate, 0.7 gm/l L-proline.
4.5 µM of TDZ and 4.4 µM of BAP are best combination of
growth regulators in which Durum Wheat produces 35
number of shoots per explant and Mature embryos of CDC
Dancer oat produces 16 shoots per explant.
Explants for direct embryogenesis include microspores,
ovules, scutellum, endosperm, embryos and seedlings.
27. Indirect Somatic Embryogenesis:-
In Indirect SE, callus is produced from explants.
Embryoids(suspensory cell to cotyledon) are
produced from callus tissue.
Explants are roots, shoots, leaf cells, anthers, seeds
etc.
Steps involved in Plant regeneration of Rice variety
through Indirect SE:-
28. a) Formation of callus b) Greening of callus c) Embryo at globular stage
d) Torpedo stage of embryo e) Cotyledonary stage and regeneration
of embryo f-g) Multiple shoot regeneration h) Complete plantlets i)
Hardening of plantlets. (Rice Variety:- Swarna)
Mondal et al., 2011
(a) (b) (c) (d)
(e) (f) (g) (h)
(i)
29. 3/27/2015
Factors affecting Somatic Embryogenesis:-
1) Genotype:-
Genetically engineered / transgenic plant
does not regenerate through SE because due
to variation.
Methylation occurs in the DNA during mitosis
then SE occurs. If Methylation occur in the
cytosine bases or H3 protein then SE get
stop.
30. 2) Explant:-
Totipotent somatic cell are used.
Immature inflorescence and Scutellar tissue of
immature seeds are used for the research. Ex:-
Triticum aestivum .
Epidermis, Procambial tissue are also produced
somatic embryo.
31. During Proembryonic
phase, 2,4-D generates
DNA Hyper methylation
so that cells in a highly
active mitotic stage.
High concentration of
auxin produces root in
somatic embryo.
2,4-D is one of the growth
regulator that produces
callus from cereals and
conc. of 2,4-D 0.1-10µM
3) Auxin:-
Polar transport of auxin
produces somatic
embryo.
Auxin concentration
will be more then
somatic embryogenesis
get stop. Ex:- Maize.
Auxin induces indirect
somatic embryogenesis
in monocots.
32. 4) Cytokinins:-
Cytokinin promote axial growth.
Cytokinin produces globular embryo from initial
embryo.
Cytokinin combination with auxin, induces somatic
embryogenesis and produce callus in cereals.
Cytokinin ratio more than auxin then it produces
Shoots.
33. 5) Gibberellic acid:-
GA promote
elongations of embryo
axis, cell division.
It synthesized of
photosynthetic
pigments in developing
somatic embryo.
It improve
photosynthetic activity,
Extra storage reserves in
vitro germination.
Hypo cotyledon are used
as explant then GA inhibit
somatic embryogenesis.
Addition of Uniconazole,
Paclobutrazol inhibit
somatic embryogenesis.
GA higher in suspensory
embryo than the proper
embryo. So GA requires
early embryo
development.
34. 6) Abscisic acid (ABA):-
ABA control tolerance and seed dormancy during
later stage of embryogenesis.
ABA induced somatic embryogenesis in high osmotic
stress and high temperature in auxin free medium.
Primary embryo contain more conc. of ABA than
secondary embryo.
Treatment of Fluridone inhibit ABA synthesis and
primary embryo does not produce secondary
embryo.
35. 7) Polyamines:-
Spermidine, Spermine and
Putrescine are added as
growth regulators and
secondary messenger.
source for plants.
Spermine act as a
Polyamines serve as nitrogen
It act as a free radical
scavengers by protecting
senescing membranes
against lipid per oxidation.
In Maize, Putrescine are
most effective with varying
concentration of GA3.
antioxidant in a medium.
It help in vegetative
growth, pollen
development, regulation of
DNA duplication,
transcription of genes, cell
division, development of
organs.
36. 8) Phytosulfokine
It modulate the culture media.
It promote somatic
embryogenesis by activating
cell division of embryogenic
cells, in presence auxin.
Phytosulfokine increases the
cell through differentiation
process.
9) Phenolic compounds:-
Phenolic compounds are inhibit
somatic embryogenesis.
4hydroxy benzyl alcohol inhibits
the globular stages.
Vanillyl benzyl ether are inhibit
the suspensor development.
Recently identification of 4
[(phenyl methoxy) methyl]
phenol involves in seed
development stills unknown.
37. Differences between Zygotic and Somatic embryo:-
Zygotic embryo
Fertilized egg or zygote.
Contain seed coat.
Produce seed.
Plantlets are healthy.
Not like to mother plant.
Propagation is low.
Somatic embryo
Sporophytic cells.
Did not contain seed coat.
Only form embryo.
Plantlets are weaker
Like to mother plant.
Propagation is high.
38. Advantages and Disadvantages of Somatic
Embryogenesis:-
Higher propagation rate. Somaclonal variation.
Suitable for Suspension
culture.
Artificial seed production.
Germplasm
conservation.
Labour savings.
39. Disadvantages
Response tissue
specific (explants).
Low frequency embryo
production.
Incomplete embryo
production.
May create unwanted
genetic variation
(Somaclonal variation).
Inability to generate
large numbers of
normal, free living
plantlets.
Plantlets are weaker.
40. Somatic Embryo Germination Media:-
MS medium containing different concentrations of
BAP (0, 1, 2, 3, 4and 5 mg/l), in combination with
different concentrations of NAA (0, 0.5, 1.0, 1.5, 2.5
and 4.0 mgL-1) were used as treatments for the
germination of somatic embryos.
Media were kept in the incubation room 25±2°C
with 16 hrs of light provided by fluorescent bulbs
and a light intensity of 16.75 µmolm-²s-¹ for eight
weeks.
Calculate the Callus induction frequency(%) and
Regeneration frequency(%).
41. 3/27/2015 Deptt of Plant Biotechnology 41
Somatic embryogenesis is an efficient plant
regeneration system.
It is potentially useful tool for genetic transformation.
Cross linking between hormone and transcription
factors is likely to play an important part in SE.
But mechanism of plant embryogenesis is unclear
and comphrensive work in future it by studying the
interaction of various factors thereby entire picture of
regulatory mechanism of embryogenesis would be
transparent.
42. 3/27/2015 Deptt of Plant Biotechnology 42
Conclusion
Indirect Somatic embryogenesis reduces the breeding
cycle.
Indirect somatic embryogenesis are used in the crop
improvement.
Indirect somatic embryogenesis are produce virus free
plants.
Indirect somatic embryogenesis are better than the
Direct somatic embryogenesis.
43. Types of In Vitro Culture
Culture of intact plants (seed and seedling culture)
Embryo culture (immature embryo culture)
Organ culture Callus culture
Cell suspension culture
Protoplast culture
Somatic Embryogenesis
Micropropagation
Somaclonal variation
44. Micropropagation
⚫Embryogenesis
Direct embryogenesis
Indirect embryogenesis
⚫Organogenesis
Organogenesis via callus formation
Direct adventitious organ formation
⚫Microcutting
Meristem and shoot tip culture
Bud culture
45. EXPLANT PREPARATION
EXPLANT : It is defined as a portion of plant body, which has been
taken from the plant to establish a culture
•Explant may be taken from any part of the plant like
root,stem,leaf,or meristematic tissue like cambium, floral parts like
anthers, stamens etc..
•Age of the explant.
• Homozygous plants are preferred.
45
50. What is Callus development ?
⚫ A callus is a blob of tissue – (mostly undifferentiated cells)
⚫ A callus is naturally developed on a plant as a result of a
wound
⚫ This callus can be left to develop or can be further
divided
51. Callus Culture
⚫Equimolar amounts of auxin and cytokinin stimulate
cell division. Leads to a mass proliferation of an
unorganised mass of cells called a callus.
⚫Requirement for support ensures that scale-up is
limited.
⚫Callus Suspension Culture
⚫When callus pieces are agitated in a liquid medium,
they tend to break up.
⚫Suspensions are much easier to bulk up than callus
since there is no manual transfer or solid support.
52. Protoplast Isolation
⚫Created by degrading the cell wall using enzymes.
⚫Very fragile, can’t pipette.
⚫The membranes are made to fuse.
osmotic shock, electrical current, virus
⚫Regenerate the hybrid fusion product.
⚫Contain genome from both organisms.
⚫Very, very difficult .
53. Use of enzymes results
in a high yield of
uniform protoplasts
after removal of cellular
debris Protoplasts can
originate from different
sources: greenhouse or
field material,
micropropagated
plants, calli,
54. Protoplast Fusion Techniques
⚫ Protoplast fuse spontaneously during isolation process
mainly due to physical contact.
⚫ Induced Fusion.
⚫ Chemofusion- fusion induced by chemicals.
⚫ Types of fusogens
⯍ PEG
⯍ NaNo3
⯍ Ca 2+ ions
⯍ Polyvinyl alcohal
⚫ Mechanical Fusion- Physical fusion of protoplasts under
microscope by using micromanipulator and perfusion
micropipette.
57. Uses for Protoplast Fusion
⚫Combine two complete genomes
Another way to create allopolyploids
⚫Partial genome transfer
Exchange single or few traits between species
May or may not require ionizing radiation
⚫Genetic engineering
Micro-injection, electroporation, Agrobacterium
⚫Transfer of organelles
Unique to protoplast fusion
The transfer of mitochondria and/or chloroplasts between species
58. Somaclonal Variation
Variation found in somatic cells dividing mitotically in culture
Ageneral phenomenon of all plant regeneration systems that involve a
callus phase
Some mechanisms:
Karyotipic alteration
Sequence variation
Variation in DNA Methylation
Two general types of Somaclonal Variation:
Heritable, genetic changes (alter the DNA)
Stable, but non-heritable changes (alter gene expression, epigenetic)
59. Somaclonal Breeding Procedures
⚫Use plant cultures as starting material
Idea is to target single cells in multi-cellular culture.
Usually suspension culture, but callus culture can work (want as much
contact with selective agent as possible).
Optional: apply physical or chemical mutagen.
⚫Apply selection pressure to culture.
Target: very high kill rate, you want very few cells to survive, so long as
selection is effective.
⚫Regenerate whole plants from surviving cells.
60. Advantages of somatic hybridization
⚫Production of novel interspecific and intergenic hybrid
Pomato (Hybrid of potato and tomato).
⚫Transfer gene for disease resistance, abiotic stress
resistance, herbicide resistance and many other quality
characters.
⚫Production of heterozygous lines in the single species
which cannot be propagated by vegetative means.
⚫Production of unique hybrids of nucleus and cytoplasm.
61. Plant germplasm preservation
⚫ In situ : Conservation in ‘normal’ habitat
rain forests, gardens, farms
⚫ Ex Situ :
Field collection, Botanical gardens
Seed collections
In vitro collection: Extension of micropropagation techniques
⯍ Normal growth (short term storage)
⯍ Slow growth (medium term storage)
⯍ Cryopreservation (long term storage
⚫ DNA Banks
62. Cryopreservation
Storage of living tissues at ultra-low temperatures (-196°C)
Conservation of plant germplasm
⯍Vegetatively propagated species (root and tubers, ornamental,
fruit trees).
Conservation of tissue with specific characteristics
⯍ Medicinal and alcohol producing cell lines
⯍ Genetically transformed tissues.
⯍ Transformation/Mutagenesis competent tissues (ECSs).
Conservation of plant pathogens (fungi, nematodes)
63. Applications:
⚫ Study of Biochemical & Physiological activities.
⚫ The effect of various hormones.
⚫Production of Secondary Metabolites.
⚫To preserve the plant species which are on red-line.
⚫Improve crop yield with regard to molecular
breeding & Genetic Engineering.
⚫To make transgenic & cis-genic plants.
64. Commercial Applications of Clonal Propagation
⚫Clonal propagation has the potential for propagation of thousands of
plantlets from a single genetic stock.
⚫Examples:
Orchids,
Potato,
Asparagus,
Strawberry, And
Various flowers or herbaceous ornamentals that set seed poorly.
⚫This may not be suitable for seeding field crops.
65. Problems in Tissue Culture
⚫ Application of protoplast technology requires efficient plant regeneration
system.
⚫ The lack of an efficient selection method for fused product is sometimes a
major problem.
⚫ The end-product after somatic hybridization is often unbalanced.
⚫ Regeneration products after somatic hybridization are often variable.
⚫ It is never certain that a particular characteristic will be expressed.
⚫ Genetic stability.
⚫ Sexual reproduction of somatic hybrids.
66. Conclusion
⚫PTC is the technique by which plant cells can be
grown in vitro sexually & asexually. By the help of this
we can study biochemical, physiological and
hormones activity.
⚫High yield, good quality of crops can be obtained.
⚫PTC , G.E. and Molecular breeding these techniques
are used to transfer the gene of same species or from
different species.
67. References
⚫Plant Tissue Culture, ELESIVISER Publishers
,Bhojwani & Rajdhan
⚫ H.S. Chawla
⚫M. S. Shekhawat
⚫Images from google search engine