Plant tissue Culture Techniques

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Propagation and establishment of high yielding banana plants (robusta sp) from shoot tip by plant tissue culture method

Propagation and establishment of high yielding banana plants (robusta sp) from shoot tip by plant tissue culture method

woody legumes and cereals. (Masoud Sheidai et al., 2010; Amornwat Srangsam and Kamnoon Kanchanapoom, 2003) The emphasis was laid to improve growth under stress condition, pest and disease resistance, improved nutritional quality, nitrogen fixation and the control of partitioning within the plant. (Wirakarnain et al., 2008) The changes in the life-style of people have shifted their consumption pattern towards nutritious foods like fruits. The production of fruits through conventional methods is not sufficient to meet the growing demand. (Juliet Akello et al., 2009; Shongwe et al., 2008) Hence, there is a need of using modern technologies like tissue culture to fill-up the gap between the demand and supply of banana seedlings. Such plants are being cultivated at select places in the state of Andhra Pradesh. It is being promoted mainly by the private companies through supplying of seed materials. At this juncture, it is important to study the performance of tissue cultured banana over that of sucker- propagated banana (Francois Lecompte and Loic Pages, 2007; Juliet Akello et al., 2007; LianJie et al., 2009; Daniel Coyneet al., 2010). Plant tissue culture techniques are essential to many types of academic inquiry, as well as tomany applied aspects of plant science (Muhammad Youssef et al., 2010). Currently, tissue-cultured plants that have been genetically engineered provideinsight into plant molecular biology and gene regulation. Plant tissue culture techniques are alsocentral to innovative areas of applied plant science, including plant
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Turmeric powder ( Curcuma longa Linn. ) as an antifungal agent in plant tissue culture studies

Turmeric powder ( Curcuma longa Linn. ) as an antifungal agent in plant tissue culture studies

Prevention or control of contamination in in-vitro plant culture has been strictly based on the use of sterile techniques. Any subsequent manipulation of the plant tissue must typically be carried out in a filtered-air environment, e.g., in a laminar-flow hood (Bottino, 1981). Endophytic or latent type contamination by bacteria and fungi (Kneifel and Leonhardt, 1992; Ryu and Holt, 1993) is an insidious process that continually threatens plant tissue culture techniques throughout the duration of the culture period. Chemical anti-microbial agents (Gilbert, 1991) such as benzyl pencillin, phosphomycin, chloremphenicol, rifamphicin (Haldeman et al., 1987; Phillips et al., 1981), nalidixic acid, etc., have been used in different crops (Chapman, 1994; Dodds, J. H., and L. W. Roberts. 1985) to avoid the contamination. These chemical antimicrobial agents generally trigger the normal physiological and chemical functions of the plant.
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Tissue Culture on Medicinal Plants –Stages

Tissue Culture on Medicinal Plants –Stages

G. superba L. is a medicinal climber, its seeds and tubers contain valuable alkaloids viz., colchicine and colchicoside as the major constituents, which are used to treat many diseases (Chitra, 2010). G. superba usually multiply by corm and seeds but due to low germination capability it restricts for the regeneration. G. superba is a commercially imperative medicinal plant which has diverse medicinal applications and eventually due to over-exploitation this plant is facing local extinction. Some time plant tissue culture techniques play a key for conservation of this plant. At present Plant tissue culture offers a valuable to overcome the problem regarding conventional propagation, and obtain disease free healthy plants (Neha Bhagat, 2011). Its seeds have poor germination and low availability, while propagation by corm also a limiting factor, making micropropagation an essential proposition in order to meet the demand for quite a huge amount for raw material by pharmaceutical industries (chaturvedi 2007). Plants have been regenerated from somatic embryogenesis, caulogenesis, direct plantlet formation and regeneration of shoot buds
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Strategies for large-scale production of commercially important banana varieties of Mizoram, India, using plant tissue culture technique

Strategies for large-scale production of commercially important banana varieties of Mizoram, India, using plant tissue culture technique

The technique of plant tissue culture has been well accepted and applied in the mass propagation of planting materials in various crops and plants. In India numerous micropropagation units are producing millions of plantlets catering the needs for the increasing demand of quality planting materials. The advantages of this technique lie in the production of plantlets that are disease free and genetically identical to the elite mother plants. Application of plant tissue culture technique is the only viable means for the large scale production of banana planting materials which is not possible through conventional propagation. The article discusses the strategies of the mass pro- duction of commercially important banana in Mizoram using plant tissue culture techniques.
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ASSESMENT OF THE VARIANTS PRODUCED THROUGH TISSUE CULTURE TECHNIQUES IN WITHANIA SOMNIFERA (L )Dunal BY RAPD PCR METHOD

ASSESMENT OF THE VARIANTS PRODUCED THROUGH TISSUE CULTURE TECHNIQUES IN WITHANIA SOMNIFERA (L )Dunal BY RAPD PCR METHOD

RAPD (Random Amplified Polymorphic DNA) analysis has been demonstrated to be sensitive in detecting variation among individuals (Sheidai et al., 2010). RAPD markers have been applied to many plant species to evaluate the clonal fidelity and genetic stability of the micropropagated plants. Somaclonal variation may be detected by using molecular marker such as RAPD and AFLP and by cytological studied (Sahijram et al. 2003).

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“A Review on Tissue Culture Studies in Eclipta alba – An Important Medicinal Plant” by Neeti Dhaka, India.

“A Review on Tissue Culture Studies in Eclipta alba – An Important Medicinal Plant” by Neeti Dhaka, India.

There is an increased focus on naturopathy and Ayurveda in recent times, which has lent prime focus upon medicinal plants. Their medicinal importance has gained attraction of firms and commercial exploiters. This has resulted in over-exploitation of these plants or their cultivation to a greater extent that is causing regional agro-ecological imbalances. Therefore, there is a need to propagate such practices that can help meet both increased demand and reduce stress on natural resources, viz., land, water and biotic material. Plant tissue culture technology is the right intervention to optimise the commercial utilisation of Eclipta alba that can maintain the biodiversity balance as well. Tissue culture protocols developed for this purpose are deployed for conserving the endangered species, rare therapeutic species and other identified species for specific purpose. The micropropagation/clonal techniques are used for mass scale multiplication from shoot tips and nodal segments. One can achieve large numbers in relatively short period of time and less space that is attractive to harness their medicinal properties commercially. Both, the yield of crop and its extracts,
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PLANT TISSUE CULTURE

PLANT TISSUE CULTURE

Embryo culture is the most successful technique of the tissue culture techniques used by plant breeders. Embryo abortion is one of the common cases that occur after cross breeding between species and sometimes after cross breeding between species. In such cases, hybrid seeds unless the embryo is isolated in early stages and grown on a diet to grow and develop into a plant. In cases where the abortion takes place at very early stages after direct fertilization, in which the embryo cannot be isolated because of its small size, the researchers isolate the whole ovule and fertilize it media appropriate until the development of the embryo. Ovules technology is also used to eliminate incompatibilities, especially in genetically divergent species. Pollen may mature at an early time before mites are ready to be vaccinated. The vaccine tube does not grow on the flowerbed due to the presence of some inhibitors in the flowering season. In vitro pollination and fertilization this problem can be overcome by isolating the oocytes and then fertilizing them with pollen from the desired plant in order to obtain the seeds of live embryos that grow later to seedlings or ovarian implants can be used in studies that relate to the appearance of fruits separation , stages of development and food needs (8).
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Thinking inside the Box: Tissue Culture for Plant Propagation in a Key Ecological Species, Andropogon gerardii

Thinking inside the Box: Tissue Culture for Plant Propagation in a Key Ecological Species, Andropogon gerardii

In this study, we showed that micropropagation was effective and efficient for a non-model grass species with implications for other ecologically important plants. We found that we were able to propagate, root, and acclimate 360 total plants over slightly less than six months. Propagation rates for the different ge- notypes of A. gerardii were higher than traditional propagation techniques in this species (e.g., rhizome propagation in the greenhouse). Hartnett (1989) found multiplication rates for A. gerardii in the greenhouse were between 0.1× and 0.4× plants per week at low density. At higher density, multiplication rates were lower [52]. Commercial cultivars grown over several years yielded 0.2× plants per week [53]. We observed greenhouse multiplication rates of 0.04×, 0.1×, and 0.05× for G2, G5, and G11, respectively, when pots were kept at 25% volumetric water content over 15 months [54]. Another study examining A. ge- rardii and A. hallii hybrids took over three years to propagate sufficient clones [55]. In other cases, studies using rhizome propagation of A. gerardii did not specify rates or duration [56] or used genetically recombined seed [44]. Overall, the rhizome propagation rates are between < 1 and 11% of the rates we observed with micropropagation. This difference is substantial, but the micropropagation advantage could vary among other non-model plant species.
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High-Tech Tissue Culture Techniques for Mass Production of Nucellar Plants of Citrus Jambhiri

High-Tech Tissue Culture Techniques for Mass Production of Nucellar Plants of Citrus Jambhiri

ABSTRACT: Despite extensive research done on nuclear embryogenesis in citrus, a major snag remains unsolved that the nuclear embryo proliferation does not lead to production of even a fraction of the number of globular embryos formed initially. During further proliferation, mostly normal differentiation of embryos gets drowned in the mass of proliferating cotyledonary structures. The present experimentation is mainly aimed to resolve this problem and produce sufficient number of nucellar plantlets per culture. The significance of nucellar embryogenesis, particularly in case of rootstocks can hardly be over emphasized. However, it is also significant in case of C. jambhiri, where commercial use of its nucellars for raising orchards is in practice. Nucellus is a somatic tissue having a genetic makeup similarly to the vegetative plant body. The culture of nucellar tissue and augmentation and induction of nucellar polyembryony have initially been the main focus of research. However, the associated extended juvenile characteristics of nucellar plantlets, if not slightly poor quality of fruit, restrict the application of in vitro nucellar embryogenesis to rootstocks only, since both such characteristics will not be of any consequence while virus-free rootstocks will be produced at a rapid rate throughout the year, which in turn is imperative for all the year round production of micrografts. When critically examined, the nucellar embryonal proliferation, i.e., embryo-to-embryo multiplication is an uncommon phenomenon. Instead, there is cotyledonary proliferation forming resette-like structures having a single radicle with pluricotyly, which is a big snag in production of nucellar plantlets.
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Application of 3D printing to prototype and develop novel plant tissue culture systems

Application of 3D printing to prototype and develop novel plant tissue culture systems

Commercial culture vessels are generally manufactured by injection moulding. However, injection moulding requires large upfront investment which makes proto- typing new vessels with this process extremely expensive [17]. Additive manufacturing (AM), or 3D-printing, is a technology in which models can be designed using a vari- ety of software and manufactured using techniques such as fused deposition modeling (FDM), stereolithography (SLA), and selective laser sintering (SLS). Due to main- stream and hobbyist adoption, 3D printers have recently become small, affordable, and user friendly. While these techniques are generally not well suited to large-scale manufacturing, they allow rapid prototyping and small scale production of specialized/customized parts. This technology allows researchers who are familiar with the problems of their system to develop problem-specific solutions that may not be known to manufacturers or may not be feasible as a commercial product. Recently, this technology has also been employed to make custom- ized labware [18–20], customized reaction-ware with reaction components printed for various chemistry appli- cations [21, 22], as well as medical simulation and educa- tion [18, 23]. While this technology has great potential to improve plant tissue culture systems for species-specific solutions, it has not yet been applied in this field.
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Biotechnological Methods and Tissue Culture Techniques for Mass Production of Nucellar Plants of Citrus limonia

Biotechnological Methods and Tissue Culture Techniques for Mass Production of Nucellar Plants of Citrus limonia

ABSTRACT: The present experimentation is mainly aimed to resolve this problem and produce sufficient number of nucellar plantlets per culture. The significance of nucellar embryogenesis, particularly in case of rootstocks can hardly be over emphasized. However, it is also significant in case of C. limonia, where commercial use of its nucellars for raising orchards is in practice. Nucellus is a somatic tissue having a genetic makeup similarly to the vegetative plant body. The culture of nucellar tissue and augmentation and induction of nucellar polyembryony have initially been the main focus of research. However, the associated extended juvenile characteristics of nucellar plantlets, if not slightly poor quality of fruit, restrict the application of in vitro nucellar embryogenesis to rootstocks only, since both such characteristics will not be of any consequence while virus-free rootstocks will be produced at a rapid rate throughout the year, which in turn is imperative for all the year round production of micrografts. When critically examined, the nucellar embryonal proliferation, i.e., embryo-to-embryo multiplication is an uncommon phenomenon. Instead, there is cotyledonary proliferation forming resette-like structures having a single radicle with pluricotyly, which is a big snag in production of nucellar plantlets.
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A critical appraisal on plant tissue culture studies in medicinally important plants

A critical appraisal on plant tissue culture studies in medicinally important plants

valuable source of different kinds of useful products like wood, medicine, fiber, firewood and timber suitable for furniture, as well as decorative plants, pharmaceutical products and aestic items. In recent years the relevance of plant tissue culture methods has gained thrust to meet the growing demands for pharmaceutical industries since the genetic diversity of medicinal plants is decreasing hap hazardously. Conventional propagation and high demand of planting material are the major constraints for the large scale multiplication of medicinal species which can be met economically and efficiently in short span of time by in vitro propagation. Micropropagation is an alternative means of propagation that can be engaged in the conservation of flora in relatively shorter time. Tissue culture is useful for multiplying and conserving the useful species which are difficult to regenerate and thus can be saved from extinction. There is an only partial advancement explored at the research level to suggest tissue culture studies in Sterculiaceae. Conclusively, this appraisal is the first of its kind which emphasizes the procedures available for in vitro propagation along with some of the remarkable achievements carried in some pharmaceutically and commercially important plants by in vitro culture techniques of plants from this family.
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The Efficient Tissue Culture System of  Orostachys fimbriata

The Efficient Tissue Culture System of Orostachys fimbriata

herb contains a variety of active substances with the effect of cooling blood and hemostasis, detoxication and re- straining wound. Therefore, O. fimbriata is used for bloody flux, partial blood, hemorrhoids blood and chronic wound which is not cured for a long time [1]. O. fimbriata is small and exquisite, with beautiful plant type and panicle with amount of flowers. It has become the “new favorite” of family gardening small fleshy plant and can also be arranged in flower bed, flower border and roof greening [2]. However, due to the great medicinal and ornamental value of O. fimbriata, wild resources are subjected to excessively exploitation and thus become threatened. In vitro culture techniques represent an excellent option for the study and conservation of rare, threatened or endangered plants [3] [4]. Therefore, the interest in using these techniques for rapid and large- scale propagation of medicinal and ornamental plants has been significantly increased [5] [6]. The present study is undertaken to establish an efficient protocol for rapid large-scale regeneration of plantlets in vitro from leaf explants of O. fimbriata.
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Tissue Culture of Momordica charantia L.: A Review

Tissue Culture of Momordica charantia L.: A Review

Advances in biotechniques, particularly methods for culturing plant cell cultures should provide new means for the commercial processing of even rare plants and the chemicals they provide .The advantage of this method is that it can ultimately provide a continuous, reliable source of natural products .The major advantage of the cell cultures include synthesis of bioactive secondary metabolites, running in controlled environment, independently from climate and soil conditions. The use of in vitro plant cell culture for the production of chemicals and pharmaceuticals has made great strides building an advances in plant science. The increased use of genetic tools and an emerging picture of the structure and regulation of pathways for secondary metabolites will provide the basis for the production of commercially acceptable levels of product. The increased level of natural products for medicinal purposes coupled with the low product yields and supply concerns of plant harvest has renewed interest in large-scale plant culture technology. Knowledge of biosynthetic pathways of desired phytochemicals in plants as well as in culture is often still in its infancy, and consequently strategies needed to develop an information based on a cellular and molecular level. These results show that in vitro plant cell cultures have potential for commercial production of secondary metabolites. The introduction of newer techniques of molecular biology, so as to produce transgenic cultures and to effect the expression and regulation of biosynthetic pathways, is also likely to be a significant step towards making cell cultures more generally applicable to that commercial production of secondary metabolites.
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TISSUE CULTURE STUDIES IN VIGNA UNGICULATA L

TISSUE CULTURE STUDIES IN VIGNA UNGICULATA L

The excised plant part called explant are at first washed with liquid detergent (5% v/v „teepol‟) then the explants are surface sterilized by 0.1% w/v mercuric chloride (Hgcl2) for a limited time (generally 10-15 minutes). The sterile explants are properly washed with sterile distilled water in the laminar air flow chamber before inoculation. The explants are incubated on the nutrient medium supplied with hormones and incubated under controlled physical condition. The suitable temperature for in vitro growth is usually 25 ± 2ºC. The cultures are and Roots.
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Evaluation of VP22 Spread in Tissue Culture

Evaluation of VP22 Spread in Tissue Culture

FIG. 4. COS-1 cells, plated on square coverslips, were transfected with the expression vector for native VP22 (600 ng). Forty hours after transfection, the coverslip was transferred to a fresh culture dish and a coverslip of control, untransfected cells was placed directly adjacent to it. The abutted coverslips were then fixed with methanol, washed in PBS, and processed for detection of VP22 by staining with AGV30. The coverslips remained directly abutted during all steps of processing. Panel 1 shows a typical field of view of the edge of the transfected coverslip nearest the abutted coverslip, while panel 2 shows a typical field of view of the edge of the untransfected coverslip directly abutting the transfected cells. Panels 3 and 4 are fields taken progressively towards the center of the untransfected coverslip, away from the transfected cells. (b) Panels 1 and 2 are as described above showing the abutted edges of the transfected and untransfected coverslips, respectively. Panel 3 shows the background from an untransfected coverslip processed separately.
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 NATURAL REMEDIES WITH POTENTIAL ANTI-CANCER ACTIVITY AND ROLE OF PLANT TISSUE CULTURE IN DEVELOPMENT OF THOSE REMEDIES: A REVIEW

 NATURAL REMEDIES WITH POTENTIAL ANTI-CANCER ACTIVITY AND ROLE OF PLANT TISSUE CULTURE IN DEVELOPMENT OF THOSE REMEDIES: A REVIEW

this review exhibit anticancer activities. Natural products offer a great opportunity to evaluate not only totally new chemical classes of anticancer agents, but also novel and potentially relevant mechanisms of action and tissue culture, increases the amount of planting material to facilitate distribution and large scale planting. In this way, thousands of copies of a plant can be produced in a short time. Micro propagated plants are observed to establish more quickly, grow more vigorously and are taller, have a shorter and more uniform production cycle, and produce higher yields than conventional propagules.
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Tissue Culture Of Rice: Problems, Progress And Prospects

Tissue Culture Of Rice: Problems, Progress And Prospects

formation and basal medium with l mg/l kin, l g/l L-proline, 300 mg/l casein hydrolysate and 20 mM L-lysine showed best regeneration. Bano et al., (2005) proved that MS medium containing 0.5 mg/l BAP and 0.2 mg/l IAA proved best for plant regeneration when seeds of Swat-II were used as explant on MS medium. Grewal et al., (2005) conducted an experiment to demonstrate the promotive effect of maltose, sucrose, proline, cefotaxime and activated charcoal on different rice varieties. It was reported that these nutrients with hormones enhances somatic embryogenesis.

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Quantitative Determination of Nitidine from Roots and Plant Tissue Culture Extracts of Toddalia asiatica (Linn ) Using HPTLC

Quantitative Determination of Nitidine from Roots and Plant Tissue Culture Extracts of Toddalia asiatica (Linn ) Using HPTLC

HPTLC method is used not only for quantification of phytochemicals [10,11] and qualitative analysis of plant extracts [12], but also for quality control of raw materials and standardization of polyherbal formulations [13,14]. Literature survey reveals that there are no reports on HPTLC determination of nitidine from the roots of T. asiatica. The present work illustrates the densitometric HPTLC method establishment and validation for quanti- tation of nitidine in roots and tissue culture extracts of T. asiatica.

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Plant Breeding Techniques

Plant Breeding Techniques

Over the last fifty years plant breeding has largely developed in response to the demands of intensive agricultural produc- tion, striving for increased yields, storability and cosmetic per- fection under a system of management based on artificial fer- tiliser nutrition and the use of pesticides. Until now, organic farmers have made use of these traditionally bred varieties but the question being asked more and more regularly is «do these varieties truly fulfil the needs of organic production?» Are the seeds and vegetative multiplication material, usually results of traditional and conventional breeding programs, adapted to the conditions of organic agriculture? And, what do consumers expect from an organic variety? Healthy, tasty, and unique prod- ucts?
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