The history of plant tissue cultivation dates back to 1902 when Haberlandt published his attempt to plant a single plant cell. However, in 1934, White was able to achieve some success in the same field when he succeeded in producing a whole plant of tomatoes from planting part of a plant root Tomato in an industrial diet, and in late 1939 a number of researchers, individually and separately, produced whole plants from the cultivation of specialized plant tissues. Both Gautheret and researcher White produced tobacco in the same way. In 1957, researchers Miller and Skoog explained that the ratio of auxin and cytokine in the food medium plays an important role in determining the nature of the growth and specialization of the plant part planted in the food medium or one. In 1958, Steward and others produced the carrot plant by developing a mass of non-specialized cells are cited by researcher Gautheret. In 1962 researchers Murashige and Skoog were able to make a quantum leap in the development of this modern science by reaching a special combination of the nutritional media for growing the tissues of the tobacco plant, which was later examined from the most famous food media used today, whether for single or double housing. In 1965 researchers, Hildebrandt and Vasil were able to use the food media (MS) for Murashige and Skoog to produce a whole tobacco plant by cultivating a single plant cell. In 1969, Nitsch found a way to grow hundreds of tobacco plants that carry half of the haploid chromosome Cultivation of pollen. In the early 1970s in several experiments, researchers were able to isolate and extract protoplast from the middle layer of leaves and then cultivate it into a food medium to produce a whole plant. The researchers also controlled the growth and specialization of protoplasts due to the varying conditions of the experiment. In 1972, Carlson and others succeeded in producing the first hybrid tobacco plant asexual when they were able to extract and synthesize protoplast from two types of tobacco and develop them on an artificial food medium to produce a whole plant. This step promised to make bells alert to the importance of this science. To carry out studies on propagation of plant tissueculture (4).
This study aimed to investigate the tissueculture and rapid propagation techniques of Orostachys fimbriata, a medicinal and ornamental herbaceous perennial herb belonging to the family Crassu- laceae. The leaves of O. fimbriata were used as explants to investigate the effects of plant growth regulators such as thidiazuron (TDZ), 6-benzylamino purine (6-BA) and 1-naphthaleneacetic acid (NAA), on the induction of callus, differentiation of adventitious buds and rooting of shoots. Our results showed that optimum callus induction medium was MS medium supplemented with 0.5 mg∙L −1 TDZ and 0.2 mg∙L −1 NAA. 1.5 mg∙L −1 6-BA and 0.2 mg∙L −1 NAA was the optimum hormone
The different explants used for the rice tissueculture include coleoptile, roots, seed derived scutellum, immature and mature embryos, leaves, stem nodes, inflorescence and anthers. Among these different explants used, seed derived scutellum serve as the best explant for the tissueculture of rice genotypes. From this study we conclude that carbohydrate, explant used, plant growth regulators, basal salts of culturing medium, culture condition and most importantly genotype of explants plays important role in successful tissueculture. MS followed by LS media with some modifications proved best for the embryogenic callus induction in diverse rice genotypes and MS and N6 media proved best for the efficient regeneration. Different carbohydrates (sucrose, glucose, maltose, sorbitol and mannitol) revealed genotype specific results in the tissueculture of rice. Likewise, different gelling agents (agar, agarose, gelrite and phytagel) revealed genotype specific results. Using different concentrations and combination of 2, 4-D, IAA, NAA, kinetin, BAP, ethylene, high frequency of callus induction was observed. The use of tryptophan, proline, casein hydrolysate, lysine, glutamine, asparginine, arginine etc. improved the frequency of callus induction. Similarly, different concentrations and combination of 2, 4-D, IAA, NAA, kinetin, BAP, 2 iP, GA R 3 R , TDZ proved best for the
While genetic analysis in Drosophila remains one of the most powerful methods to determine the function of a particular protein involved in cell division (reviewed in ref. 28), this is not always possible due to the lack of mutations for particular genes or due to the complexity of phenotypes observed with certain hypomorphic alleles. In those cases, RNAi in Drosophila tissueculture cells can be an excellent alternative for the study of gene function. It is important to note that analysis of fixed material can lead to ambiguous interpretations since each fixed sample represents a single time point in what is a highly dynamic process. Live cell imaging would be of great utility for the study of mitosis on RNAi-depleted cells. In fact, this is now routinely performed for RNAi experiments using human cells. In those cases, cells are transfected with siRNAs or suitable vectors, then blocked in S phase with elevated levels of thymidine. Upon release from the thymidine block, cells are filmed as they enter mitosis. Unfortunately, this approach has yet to be developed for Drosophila cultured cells, where so far in vivo analysis of mitosis has been restricted to embryos and primary cultures of larval neuroblasts (29-30).
our present study: - (1) In identifying the sterilization protocol for sugarcane explant (var. CoJ 85), our experience with the followed protocol is satisfactory. (2) We have also observed that the explants have to be subcultured for 3-4 times (in 7-10 days interval) for at least 4 weeks in solid medium before transfer to the liquid medium for further establishment. The media combination used for initial establishment phase is MS Basal + 0.5mg/lit BAP + 0.5 mg/lit KN + 30 gm/lit Sugar + 8.0 gm/lit agar. (3) At the multiplication stage, we found that two combinations MS + 1.0mg/lit BAP + 1.5mg/lit Kn + 30gm/lit Sugar and MS +0.5mg/lit BAP + 0.5 mg/lit Kn + 30 gm/lit Sugar has shown best results for multiplication rate. However, at this stage, shoot elongation has been found to be maximum in MS + 1.5mg/lit BAP and 0.5mg/lit KN + 30 gm/lt. (4) Significant rooting response found in media combinations having MS + 5mg/lit NAA + 70 gm/lit sugar and as compared to the media with same conc of Auxin but reduced sugar (30gm/lit) has a reduced response for rooting. (5) We have transplanted a few plants for hardening but due to time constraints we could complete the experiments. To standardize a Successful Tissueculture protocol, successful hardening of microshoots is very essential and further experiments are needed. (6) For callus induction, MS + 2,4-D 4mg/lit is found to be satisfactory, however, we could not experimented further with the callus growth. (7) In order to ascertain the true to type character of the Tissueculture raised plantlets through axillary shoot proliferation, it is essential to check at the genomic level, it is felt that further experimentation is needed towards achieving the result.
Abstract: Plant tissueculture is the technique to culture plant cells or tissues under controlled aseptic conditions on a synthetic medium .It has value in basic research like cell biology, genetic transformation studies &biochemistry for the production of medicinally valuable secondary metabolites. Besides this also has commercial application. This review work outlines the work done on the tissueculture of Momordica charantia L. Momordica charantia L. commonly known as bitter melon/gourd, a member of Cucarbitaceae, is a slender, tendril climbing, annual vine. Bitter melon is a common food item of the tropics and is used for the treatment of cancer, diabetes, AIDS and many ailments. It is a potent hypoglycemic agent and its hypoglycemic actions for potential benefit in diabetes mellitus are possible due to at least three different groups of constituents in bitter melon. These include alkaloids, insulin like peptides, and a mixture of steroidal sapogenins known as charantin. Clinical studies with multiple controls have confirmed the benefit of bitter melon for diabetes. Alpha and beta momarcharin are two proteins found in bitter melon, which are known to inhibit the AIDS virus.
Tissueculture has wide applications in research as it offers numerous benefits over the traditional propagation methods. For example, a single explant can be multiplied into several thousand plants in less than one year. Once established, actively dividing cultures are a continuous source of microcuttings, which can result in plant production under greenhouse conditions without seasonal interruption. The rate of growth is much faster due to the addition of exogenous hormones in vitro compared with traditional methods. The environment in culture provides nutrient and sterile conditions to avoid from other organism and the competitor of nutrient uptake (Lineberger, 2003; Slater et al., 2003).
Fragments of synovium from patients with rheumatoid arthritis survive in defined tissueculture medium in the absence of added serum and, after 3-4 days, release into the medium enzyme capable of degrading undenatured collagen. Maximal activity is observed at pH 7-9 but the enzyme is inactive at pH 5. At temperatures of 20° and 27°C, collagen molecules in solution are cleaved into 3/4 and 1/4 length fragments with minimal loss of negative optical rotation, but with loss in specific viscosity of approximately 60%. Above 30°C the fragments begin to denature and denaturation is complete at 37°C. If the enzyme is not inhibited at this stage the large fragments are broken down further to polypeptides of low molecular weight. Reconstituted collagen fibrils and native fibers at 37°C are cleaved to the low molecular weight fragments, although the fibrils are resistant to breakdown at lower
Tissueculture evolved on the basis of cell pluripo- tency and cell differentiation theories. By using plant tissue cells from pollen, embryo, seed, stem segments, buds, and apical tissues, and with the induction of growth regulators in a sterile state, cultured cells can grow into fully grown plants. Due to asexual reproduction, the ex- cellent traits of the mother plant can be well retained in the offspring [14,15]. The operating processes of the production of healthy seedlings using tissueculture tech- nology can be divided into the follow stages:
virus grew to high titers without inducing fusion. As the pre- dicted amino acid sequence of the two viral glycoproteins was unchanged, this suggests that the ability to replicate in Vero cells was not at the receptor level. This was substantiated by showing that the virus did not use CD46 as a receptor, which is in agreement with a lack of Tyr at position 481 for CD46 usage (7). However, as Vero cells do not express SLAM, the mech- anism by which the virus infects the cell remains to be deter- mined. In human immunodeficiency virus infections, the virus can incorporate cellular proteins which can attach to their corresponding ligands on host cells (reviewed in references 13 and 15). However, in our system, anti-Vero sera failed to neutralize G954.V10 infection of Vero cells (T. F. Wild, un- published results). In the present study, we have analyzed a number of the parameters displayed by wild-type MV when adapted to tissueculture. MV can enter such cells and follow- ing a period grows to high titers without giving classical fusion. These observations may be relevant to the in vivo situation in which viral antigen is found in cells which are not known to express the MV wild-type receptor SLAM. It remains to be established if these cells become infected by a mechanism similar to the present studies or via a third MV cell receptor.
Cowpea is a dicotyledonous plant belonging to the family Fabaceae and sub-family, Fabiodeae. Cowpea is often called as "black-eyed pea" due to its black- or brown- ringed hylum. Vigna is a nutritious source of food, its grains contain about 25% Protein, especially rich in folate, potassium, iron, magnesium and the essential amino acids lysine and tryptophan. It is also used as animal fodder, cover crop and green manure. Cowpea is often called as "black-eyed pea" due to its black- or brown-ringed hylum. Vigna originated in West Africa India, Nigeria accounts for 70% of the world‟s production of cowpea beans. The plant tissueculture is also proving to be rich and novel sources of variability with a great potential in crop improvement without restoring to mutation or hybridization. The plant with long seed dormancy can be raised faster by in vitro clonal propagation. In the present study shooting was initiated from nodal and shoot tip explants on MS medium containing BAP alone used as phytoharmone source. Rooting in the form of organogenesis was obtained from leaf explants in the presence of NAA alone as phytoharmone.
Plant tissueculture, particularly micropropagation techniques can be applied to clone a large number of an endangered plant with minimal damage to the natural populations (Carson and Leung, 1994; Faisai et al., 2007; Chaudhuri et al., 2008; Mohammadi-Dehcheshmeh et al., 2008). Although the large number of plants regenerated in the tissueculture laboratory can be introduced back to the natural environments, there might be some unforeseen potential risks associated with monoculture practice. Clearly this is not desirable. Therefore, the main utilily of the clonal plants is for experimental studies as there would be little or greatly reduced variability due to plant materials. They can be deployed to gain clearer insights into all aspects of the biology of the endangered plants. These are pre-requisites to development of strategies to manage them in relation to the whole spectrum of potential threats including climate change. Plants regenerated following this in vitro selection scheme might be genetically improved plants with an expanded capacity to cope with or even thrive in drier conditions that are projected to result from climate change. Therefore using this non-controversial, non-genetic engineering approach could be an option to ensure the sustainability of the endangered plant (Gopal et al., 2008). People’s participation in conservation of rare and endangered medicinal plants like G. superba will also be very useful. There is a pressing need to conserve the plant by in situ and ex situ multiplication in general and micropropagation in particular so as to meet the ever increasing demand from the industries (Kapai et al., 2010).
FIG. 2. (A) MA104 cells 15 min after fusion by TA BRV. Fused cells were visible as doublets and triplets (arrows), with apparently no separating membranes. For fusion, TA BRV was adsorbed to a single-cell suspension of cholesterol-treated MA104 cells at 4 8 C for 15 min. Cells were pelleted, incubated at 37 8 C for 15 min to allow fusion, plated in six-well tissueculture dishes, and immediately photographed. (B) Control MA104 cells which had undergone the fusion procedure but without added virus and were photographed at the same time point as those in panel A. Most cells were singlets, and when they were present as doublets, a clear plasma membrane was visible between the two cells (arrow). Magnification in panels A and B, 3 218. (C and D) Two types of cell-cell fusion were visible in TA BRV-fused MA104 cells photographed 3 h postfusion, after the cells had attached and spread on a tissueculture plate. In panel C, fused cells have connecting tubes of cytoplasm (arrows) with no obvious intervening membrane separating the cells. Panel D shows small syncytia with multiple nuclei (arrows). Magnification in panels C and D, 3 218. (E and F) High- and low-power magnifications of control MA104 cells which underwent the fusion procedure but without added virus. Cells were fixed as a pellet 1 h after being brought to 37 8 C to induce fusion. Panel E shows clear separation between adjacent cells. Magnification, 3 5,000. Panel F, a higher-power magnification of the boxed area in E, shows a distinct bilayered plasma membrane. Small circles between cells are cross-sections of microvilli (arrow). Magnification, 3 20,000. (G) High-power magnification ( 3 75,000) of the region where two MA104 cells have fused. Osmophilic areas (dark patches) of unknown origin often are present at cell-cell boundaries. Here the osmophilic areas are still visible but no plasma membrane is detectable. Note the scattered pieces of membrane embedded within the cytoplasm (arrows), which probably
Int 1 De\' 1Iiol 15 2S9 2~3 (1991) 259 Peroxidase as a developmental marker in plant tissue culture MARIJANA KRSNIK RASOL Department of Molecular Biology, Faculty of Science, University of Zagreb, Rep[.]
8 Plant tissue was placed in a 55 mm plastic Petri dish, along with 0.5 cm 2 Pisum sativum L. ‘Ctirad’ leaf tissue, an internal standard known genome size of 2C = 9.09 pg. Four hundred μl of extraction buffer (CyStain UV Percise P Nuclei Extraction Buffer, Partec, Munster, Germany) was added to the Petri dish. Material was finely chopped for 30 to 60 s with a razor blade and incubated for approximately 30 s (no more than five minutes). The suspension was filtered through Partec 50 µm CellTrics disposable filters into a sample tube. Nuclei were stained with 1.6 mL of 4’, 6-diamindino-2-phenylindole (DAPI). Samples were again incubated for 30 to 60 s. Nuclei were analyzed using a flow cytometer (Partec PA II) with the blue florescence channel. Genome sizes of samples were calculated as: (mean florescence of unknown/mean florescence of known standard) * genome size of known standard.