Application Of Tissue Culture Techniques Research Articles

Influence of Cytokinins, Dark Incubation and Air-Lift Bioreactor Culture on Axillary Shoot Proliferation of Al-Taif Rose (Rosa damascena trigintipetala (Diek) R. Keller)

Rose is a widely favored floriculture crop that is commercially propagated through the application of tissue culture techniques. Here, we report an effective method for axillary shoot proliferation in Al-Taif rose, an important cultivar for rose oil industry. Stem nodes were excised from an adult donor Al-Taif rose shrub and cultured for 4 weeks on Murashige and Skoog’s (MS) medium supplemented with 6-benzylaminopurine (BAP) or gibberellic acid (GA3) at 0 and 3 mg·L−1 to induce the sprouting of axillary shoots. Al-Taif rose shoots were cultured in vitro for 6 weeks on MS medium fortified with different concentrations of cytokinins, light/dark incubation and different culture types (gelled and liquid/bioreactor culture). The culture conditions that were applied had a noteworthy impact on the responses of Al-Taif rose shoot proliferation. The supplementation of the medium with 6-benzylaminopurine (BAP) resulted in an augmented rate of shoot proliferation in comparison to other cytokinins. Additionally, dark incubation limited foliage growth, leaf yellowing and abscission and favored shoot proliferation compared with light incubation. Liquid culture using bioreactors provided higher axillary shoot proliferation and growth as compared with gelled culture. A continuous immersion system with a net provided the highest axillary shoots (four shoots per explant) and shoot length (16.5 cm), whereas an immersion system without a net provided the highest fresh weight of axillary shoots (499 mg per explant). These findings will improve commercial propagation and contribute to the rose oil industry of Al-Taif rose.

In vitro multiplication of Eugenia calycina Cambess. (Myrtaceae) under the influence of BAP and NAA

Eugenia calycina Cambess. (Myrtaceae) is a fruit species native to Brazil indicated for the recovery of degraded areas. Generally its propagation occurs by seeds, but leads to an increase in the juvenile period of the plants, problems in maintaining seed viability and pathogen incidence. However, with the application of tissue culture techniques it is possible to produce seedlings in large scale, uniformity of the plants obtained and reduction in the period of seed germination. Thus, the objective of this research was to define the best medium for in vitro multiplication of E. calycina. Nodal segments of 1 cm, extracted from E. calycina plants already established in vitro, were used as explants. The treatments consisted of different concentrations of BAP (0; 0.5; 1 and 2 mg L-1) and NAA (0; 0.5; 1 and 2 mg L-1). The experimental design was completely randomized, in a 4x4 factorial scheme, with four concentrations of BAP and four concentrations of NAA, with four replications and three tubes per replication. After 90 days, shoot number, length and fresh mass of the aerial part, production and fresh mass of callus were evaluated. The MS medium plus 2.0 mg L-1 of BAP is ideal for in vitro multiplication of E. calycina.

IN VITRO PROPAGATION OF PLUM ROOTSTOCK DOCERA 6

This paper describes research on the application of tissue culture techniques to the micropropagation of interspecific rootstock ʹDocera 6ʹ. The experimental work was carried out in the period 2017-2018, in the in vitro propagation laboratory of the Fruit Growing Institute Plovdiv. Аxillary buds were employed as initial explantеs in two different seasons (March-May; September-October). The action of the mineral medium was studied in the multiplication stage. The best result was obtained on LS medium included BAP 0.5 mg/l and IAA 0.05 mg/l. Тhe obtained average multiplication rate is 3.08. The concentration of auxin applied to the basal medium influence the quality of the root system Treatment with high concentrations of IBA added to the rooting medium gives the best results (V5). Тhe influence of the season on growth and development of micropropagated ʹDocera 6ʹ rootstock during ex vitro acclimatization is also part of our research. The spring acclimatization gives better results than the autumn.

Application of tissue culture techniques in potato

Application of tissue culture techniques in potato

Iron Chelation and Iron Reducing Activity of Tissue Cultured and Tissue Culture Derived Mentha Spp

The antioxidant potential of herbs varies greatly with varietal differences, cultivation conditions and environment . In the present study, the effect of maturity of tissue cultured (TC) and tissue culture derived (TC0 andTC1) different Mentha species on iron chelation and iron reducing potential was evaluated. Mentha sps. selected for the study were Mentha piperita (PPR 611), Mentha arvensis (SH) and Mentha spicata (SPR-8). The variation in total phenolic and flavonoid content in tissue cultured and tissue culture derived plantlets can be due to culture induced variations in biosynthetic pathways. Also, the iron reducing and iron chelation property was found to be better in Mentha Spicata sps. Iron reducing property increased with increase in growth period, however the iron chelation decreased with increase in development period for three months. Also the application of tissue culture techniques in conserving and augmentation of secondary metabolites in Mentha sps was evident from results.

Protocol: Ex Vivo Culture of Mouse Embryonic Mammary Buds

The explant culture techniques of embryonic tissues allow continuous monitoring of organ growth and morphogenesis ex vivo. The effect of growth factors and other soluble molecules can be examined by applying them to the culture medium. Relatively few studies have reported application of tissue culture techniques to analysis of embryonic mammary glands. Here we describe a protocol for murine mammary rudiments that permits ex vivo development up to branching stage.

Tissue culture of rubber: past, present and future prospects

In rubber, the current propagation method of grafting on to unselected seedlings, maintains intraclonal heterogeneity for vigour and productivity and hence a great improvement may be expected by using micropropagation in vitro. Micropropagation with nodal and shoot tip explants derived from seedlings are possible as with mature clonal explants. The major problem in using clonal material from mature trees of Hevea is the failure to produce an adequate tap root system necessary for tree stability, and the poor response to culture conditions. The latter problem has been over come to a significant level by in vitro micrografting. Recently, there has been an increasing interest in the induction of somatic embryogenesis in Hevea using different explants, media compositions and conditions, especially for use in genetic transformation studies. Successful somatic embryo formation and plant regeneration have been reported by a few researchers in different countries using limited genotypes of Hevea. Attempts to induce somatic embryos from high yielding Sri Lankan clones have not been successful until the late 1990s. The frequency of somatic embryo induction was found to be very low and non-synchronous, its germination remained very difficult and thus Hevea embryogenic system needed further investigation. There is no large scale commercial application of tissue culture techniques for mass propagation of clonal Hevea as yet, either by microcuttings or by somatic embryogenesis. However, there is sufficient progress at research level to suggest that tissue culture of Hevea can and should be further developed. Key words: Hevea brasiliensis, micropropagation, microcuttings, somatic embryogenesis doi: 10.4038/cjsbs.v36i2.486 Cey. J. Sci. (Bio. Sci.) 36 (2): 116-125, 2007

APPLICATION OF RECENT BIOTECHNOLOGIES IN THE CONSERVATION OF RARE FRUIT SPECIES FROM DEVELOPING COUNTRIES

ISHS XXVII International Horticultural Congress - IHC2006: International Symposium on Seed Enhancement and Seedling Production Technology APPLICATION OF RECENT BIOTECHNOLOGIES IN THE CONSERVATION OF RARE FRUIT SPECIES FROM DEVELOPING COUNTRIES

Micropropagation enables the mass propagation and conservation of Dyckia distachya Hassler

The Atlantic Forest has a rich biological diversity and elevated levels of endemism in which bromeliads are frequent, but endangered. Tissue culture techniques raise high hopes for the rescue, mass propagation for commercial aims or the conservation of natural populations threatened with extinction. The present study evaluated aspects of seed-based D. distachya micropropagation. A high rate of in vitro regeneration (78.93 shoot gram -1 ) was obtained in response to liquid MS culture medium supplemented with Benzylaminopurine 2 mM after 120 days in culture, or supplemented with Naphthaleneacetic acid 2 mM, BA 4 mM and Paclobutrazol 6 mM (133.58 shoots explant -1) after 142 days in culture. Not less than 1.5 cm long shoots were successfully acclimatized to ex vitro conditions. D. distachya is responsive to the application of tissue culture techniques for mass propagation and conservation purposes. The use of seeds as initial explants allows the maintenance of the genetic diversity observed in the natural populations and mass regeneration is a promising technique for the conservation of this endangered bromeliad.

Glial Precursor Cells in the Adult Human Brain

Oligodendrocytes, the glial cells responsible for laying down and maintaining myelin sheaths in the central nervous system, were first described only 75 years ago. The lineage of these cells, and its relationship with that of the second type of macroglia, the astrocyte, was much studied in vivo and in situ in the rodent over the next 60 years. In the early 1980s, progress in oligodendrocyte biology was markedly amplified by the application of tissue culture techniques–-not without some element of controversy, although this is now largely resolved. Oligodendrocytes have always been given more attention than many other cells as a consequence of their role as a key target in human demyelinating diseases; in fact, few studies of rodent oligodendrocytes fail to draw conclusions regarding multiple sclerosis. Now, however, techniques for studying human glia and their lineage more directly have emerged, and differences in rodent and human oligodendrocyte biology are becoming apparent. It is increasingly clear that some caution must accompany the uncritical extrapolation of rodent experimental data to human oligodendrocyte biology and, indeed, to human disease.

Micropropagation of juvenile and adult Quercus suber L.

This paper describes research on the application of tissue culture techniques to the micropropagation of cork oak (Quercus suber L.), a forest species of ecological and industrial importance in the Mediterranean area. Apical buds and nodal stem segments were employed as initial explants. Their origins were young seedlings, stump sprouts and sprouts formed on cuttings collected from old trees.

The Application of Tissue Culture Techniques in Lupin Breeding

The Application of Tissue Culture Techniques in Lupin Breeding

Retinoblastoma cells in tissue culture.

This review summarizes and discusses research on retinoblastoma (Rb) cells in tissue culture. Retinoblastoma is an intraocular tumor of early childhood which is believed to originate from the primitive multipotential neuroectoderm of the optic cup region. The application of tissue culture techniques to the study of Rb cells permits detailed studies of the biology of this tumor. Classic studies have primarily focussed on growth and metastatic potential of Rb cells. Y-79 Rb cells, for example, have a short doubling time in vitro as well as aggressively growing in the anterior chambers of athymic 'nude' mice. Such active growth may result from secretion of a Retinoblastoma Derived Growth Factor (RDGF) by the cells. Several natural agents have now been shown to halt Rb cell growth in vitro. Among these are the fatty acid, butyrate, and two retinoids: retinol and retinoic acid. Interestingly, the retinoids have different mechanisms of action. Cultured Y-79 and WERI cells appear to be multipotential in that they exhibit both neuronal- and glial-like characteristics. Natural agents such as cyclic AMP and butyrate can induce the cells to differentiate along either neuronal or glial cell lines as assessed morphologically and immunocytochemically. Of interest is that combination of agents such as butyrate and laminin, an extracellular attachment protein, yield totally different morphologies, in this case, pigment epithelial in nature. Tissue culture studies thus not only show the primitive, multipotential nature of the Rb cells but their great plasticity as well. Such studies are also useful in elucidating the multiple factors (e.g., substrata and soluble agents) which code for normal retinal development from embryo to adult.

Articular cartilage preservation and storage. I. Application of tissue culture techniques to the storage of viable articular cartilage.

Articular cartilage slice explants were stored under various conditions, including freezing-thawing at various rates by using dimethyl sulfoxide (DMSO) as a cryoprotective agent, incubating in standard tissue culture medium (MEM Eagle:NCTC 135:15% fetal calf serum) in 5% CO2 and air at 4 degrees, 21 degrees, and 37 degrees C, and incubating in standard tissue culture medium containing 200 micrograms/ml alpha-tocopherol (vitamin E) at 37 degrees C after first ascertaining a dose-response curve of vitamin E. Results indicated that articular cartilage slice explants did not survive freezing or storage at 4 degrees and 21 degrees C as measured by 35S uptake. When stored at 37 degrees C in standard tissue culture in 5% CO2 and air, the slice explants remained viable for up to 60 days. The addition of alpha-tocopherol to the medium resulted in significantly less release of previously incorporated 35Sin stored cartilage slices and significantly less reduction of the amount of hexosamine present in the stored explants. alpha-Tocopherol in the medium also preserved safranin O staining. Thus, the application of tissue culture techniques to the storage of articular cartilage made it possible to preserve cartilage slice explants in a viable, biochemically "normal" state.

Application of Tissue-Culture Techniques to the Study of Corticoidogenesis in the Mammalian Adrenal

Application of Tissue-Culture Techniques to the Study of Corticoidogenesis in the Mammalian Adrenal

The application of tissue-culture techniques to the chromosomal analysis of Bos taurus

Until 1960 all the chromosome counts performed on cells of this species were done on histologically prepared specimens of testicular tissue. In January 1960, Chiarelli et al. published the results of their experiments to grow renal tissue in culture. Their specimens were obtained from male and female calves. They confirmed that 2n = 60 and described the chromosomes as being all acrocentric except the X chromosome, which was submetacentric.The purpose of the work described below was to try to apply the peripheral-blood culture technique of Moorhead to bovine blood, and to attempt to grow bovine muscle cells in a tissue culture.