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To study the influence of genetic background on the transformation and regeneration of cultivated tomato plants, hairy root lines of tomato (Lycopersicon esculentum) were obtained by inoculating the hypocotyl explants of three tomato cultivars with the Agrobacterium rhizogenes strain DCAR-2, which harbors the pBI-121 binary vector. The Ri-T-DNA transformation into the plant DNA was confirmed by both of mikimopine and GUS assay analyses. The regeneration efficiency from hairy root explants was assessed. The data indicated that white embryonic calli were formed within two weeks in the presence of 2 mgl-1 2, 4-D plus 0.25 mgl-1 kinetin. Adventitious shoots emerged from the embryonic callus in the presence of 1 mgl-1 GA3 along with 0.5 mgl-1 NAA. The regeneration frequency was higher in the cultivar UC-97, followed by Momotaro and then Edkawi. Molecular confirmation of the integration of the GUS gene into the hairy root-derived plants genomes was done via PCR using GUS-specific primers and also using Southern blotting analysis. Our data shows that regeneration is possible from hairy roots of the cultivated tomato and this system could be used to produce transgenic tomato plants expressing the genes present in Agrobacterium rhizogenes binary vectors.
Isatis concstricta Davis (an endemic plant of Turkey) suffers from low propagation rates under natural conditions and is threatened due to fast unplanned urbanisation. The study compared the effects of variants of BA + NAA on shoot regeneration on leaf and petiole explants excised from one week old in vitro regenerated seedlings. MS medium containing 1 mg L–1 BA + 1 mg L–1 NAA induced maximum proliferation on petiole and leaf explants with 13.33 and 12.75 shoots per explant repectively. However, leaf explant induced shoots were sturdy and healthier compared to petiole explant induced shoots. These shoots were rooted on MS medium containing 0.5 mg L–1 IBA and the plants were acclimatized in peat moss and sand (v/v). They grew to flowering under ex vitro conditions. This system of regeneration is advantageous for conventional propagation and the results will help in establishment of a powerful and meaningful micropropagation system for I. constricta.
Regeneration efficiency from three different regions of cotyledonary explants was examined in six sunflower inbred lines. Proximal, middle and distal regions from seedling cotyledons were cultured on regeneration medium supplemented with growth regulators. Plant regeneration by direct organogenesis was observed after four weeks. Significant differences among inbred lines were found for regeneration percentage and average number of shoots per total explants. Also a decreasing regeneration capacity was observed from proximal to distal sections for all inbred lines. Regeneration ability from cotyledonary explants in this species is strongly influenced by the genotype and by the region from which the explant was obtained. The distance to the cotyl edonary node plays a preponderant role in the expression of shoot forming capacity. Shoot differentiation via seedling cotyledons depends upon the presence of the proximal region of cotyledon regardless of the genotype.
A protocol for the induction of regeneration from leaves of Helichrysum italicum was established. Calli were found to form on the basal medium only when it was supplemented with thidiazuron (TDZ) alone or in combination with naphthalene acetic acid (NAA), with a percentage ranking of at least 80%. The hormone-free medium showed the highest percentage of shoot regeneration (62%) even though no callus formed. AFLP markers were employed to verify tissue culture-induced variation in the regenerated plantlets obtained by direct shoot regeneration or the indirect shoot regeneration process (callus formation). Seven out of the eleven AFLP primer pairs yielded polymorphic patterns. The average number of fragments per primer pair was 64.1. Singletons were represented by 12 (2.7%) fragments. Student’s T-test was performed both on the average number of shared fragments and on the nucleotide diversity, and no significant statistical difference was observed between the two regeneration treatments.
Badano regenerację pędów z fragmentów bulw Gloriosa rothschildiana O’Brien uzyskanych in vitro. Fragmenty bulw zawierające pąk wyłożono na zmodyfikowaną pożywkę MS (Murashige i Skoog 1962) uzupełnioną BA lub TDZ w stężeniach: 0,5, 1, 2,5, 5, 10 mg·dm⁻³. Jako kontrolną zastosowano pożywkę bez substancji wzrostowych. Po 8 tygodniach pędy pasażowano na pożywkę zawierającą BA 5 mg·dm⁻³ i IAA 0,1 mg·dm⁻³ i kultywowano przez kolejne 10 tygodni. Największą liczbę pędów w etapie inicjalnym uzyskano na pożywce z dodatkiem 10 mg·dm⁻³ TDZ. Pożywka inicjalna miała wpływ na współczynnik namnażania i wzrost pędów w kolejnym pasażu. TDZ w stężeniu 10 mg·dm⁻³ wywierał najsilniejszy wpływ następczy.
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