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Micropropagation of Calycanthus fertilis

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Calycanthus fertilis Walt. is a shrub belonging to the family Calycanthaceae, has great potential as ornamental. In the literature there are no reports on methodpropagation of this shrub in in vitro cultures. Therefore, the aim of this study was a development of the method micropropagation of Calycanthus fertilis Walter. Shoot explants of the size 1 cm, with an apex or node with lateral meristems were placed on the media with mineral composition according to MS and WPM supplemented with BAP (from 0.5 to 2.0 mg·dm–3) and TDZ (from 0.1 to 0.5 mg·dm–3). BAP turnedout to significantly increase initiation frequency whereas TDZ inhibitedthe formation of adventitious shoots andcausedexplant death. The highest percentage of initiated explants were found in shoot fragments placed on WPM medium supplemented with 1.0 mg·dm–3 BAP. Primary explants which initiatedgrowth were transferredon the proliferation media containing WPM macroand microelement, with the addition of different cytokinin: BAP (from 0.5 to 2.5 mg·dm–3), KIN (1.0 to 5.0 mg·dm–3) or TDZ (from 0.1 to 0.5 mg·dm–3). Calycanthus multiplication in vitro shouldbe conductedon WPM media with 1.0 mg·dm–3 BAP. Proliferatedshoots were placedon the WPM rooting medium supplemented with auxins: IBA, IAA or NAA at the concentration from 0.1 to 2.0 mg·dm–3. Maximum rooting was obtained on WPM medium supplemented with 0.5 mg·dm–3 IBA. To sum up, it shouldbe statedthat an efficient method of micropropagation of Calycanthus fertilis Walt. has been developed.
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65 years of in vitro culture in Poland

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This paper is a short review of historical development of the tissue culture method in Poland. Similarly to the rest of the world, in vitro technology in Poland has progressed in many directions simultaneously. Its main fields are closely interconnected by natural sequences of biological processes and integrade one into another. The best results, driven by the prospects of practical applications, have been achieved within the areas of meristem culture and micropropagation of valuable genotypes, somaclonal variation, production of haploids and DH plants, somatic embryogenesis, in vitro culture of embryos and ovules, improvement of plant disease resistance, somatic hybridization and transformation of plants. Due to the fact that in vitro domain is a very broad science the authors are aware that this review might not fully cover all the scientists working with tissue culture and their achievements.
This paper describes analysis of free and total (also liberated by hydrolysis) ellagic acid (EA) content in in vitro cultured Rubus chamaemorus and in leaves of intact plants as a con­trol. The content of EA was determined by high performance liquid chromatography (HPLC). The analysis revealed that the content of free EA in the shoot cultures was only 2.6 X less than in intact leaves and that callus tissue is also able to synthetize small amounts of EA.
Vernonia herbacea is a native species from the Brazilian Cerrado that accumulates about 80 % of inulin-type fructans in the underground reserve organs, the rhizophores. This work aimed at establishing a protocol for in vitro culture of V. herbacea, using seeds (achenes) and leaf discs as explants. Following germination and seedling growth, stem nodes from 6-month-old in vitro germinated plants were isolated and incubated on culture medium free of growth regulators for plant propagation and rhizophore formation. Fructan content and composition were evaluated in leaves, stems, roots and rhizophores from plants grown in vitro and compared with those of greenhouse-grown plants, in order to evaluate inulin production in vitro. Fructan contents of aerial organs and roots from in vitro plants were higher, compared with greenhouse plants, while in rhizophores, the opposite was observed. High performance anion exchange chromatography/pulsed amperometric detection profiles revealed the presence of the inulin homologous series in the aerial organs exclusively for in vitro plants, while in roots and rhizophores, this series was detected in plants grown in both conditions. These results indicate a modification in the source/sink ratio, leading to changes in the distribution of carbohydrates in in vitro plants. The leaf disc cultures on medium supplemented with indole-3-butyric acid induced the formation of roots (0.24, 0.49 µM) and friable callus (2.46 µM), while 6-benzylaminopurine (from 1.1 through 4.43 µM) induced compact callus. However, no shoot formation was observed. The use of seeds allowed the establishment of a protocol for in vitro culture and provides a model system for a better understanding of fructan metabolism in V. herbacea.
The present study was aimed to develop a cost-effective and efficient protocol for mass propagation of high-quality seedlings through tissue culture by using seaweed extract as biostimulants instead of synthetic chemicals. The nodal explant of field grown W. somnifera estabilised on Murashige and Skoogs medium (MS) and Gamborg B5 medium supplemented with six concentration of 2,4 D. The percentage of culture response from the nodal explant ranged from 44 to 80 and 3.0 mg l-1 2,4 D found to be best for callus induction. MS media containing different concentration of seaweed extract (10, 20, 40, 60, 80 and 100 %) were tested individually for shoot induction. The medium supplemented with 40 % seaweed extract exhibited maximum number of shoots with about 8.6 shoots/ callus and 80 % seaweed extract exhibited 4.3 shoots/ callus. It is evident from this study that seaweed extracts can be used as substitute for synthetic growth hormones for micropropagation of medicinally important plant W. somnifera for clonal propagation and conservation.
Stevia is a plant attracting attention due to its capability to synthesize a group of chemical compounds with sweet taste, i.e. steviol glycosides. Steviol glycosides are successfully applied as a natural sweetener, and some of them have also therapeutic properties. This paper presents available information on the use of stevia plant tissue cultures with the focus on their potential application in food industry. Detailed analysis was done concerning the research employing in vitro culture techniques and the use of them in biosynthesis of secondary metabolites of high importance for the food industry. Both established achievements and most recent publications on stevia were used for assessment of practical applications of the aforementioned techniques and prospects for their development.
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