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The regenerative ability of explants from various organs of Alstroemeria plants was investigated. Rhizome apical and axillary tips cultured on the Murashige and Skoog medium with BA - 2 mgl-1 and NAA - 0.5 mgl-1 were the best among the tissue tested as initial explants. Five weeks after isolation the rhizome with 1-4 upright growing shoots were obtained. The types of rhizome explants influenced development and growth of lateral rhizomes and upright growing shoots. There were no significant differences in number of roots formed on various kind of rhizome explants. Rooting was strongly influenced by NAA. Subapical segments of vegetative stem, segments of flower pedicels and parts of ovary did not regenerate rhizome or roots but occasionally callus was formed on the medium with kinetin - 2 mgl-1 and NAA - 2 mgl1. Segments excised from vegetative stem sporadically developed roots on the medium with NAA or IBA in concentrations 3 and 9 mgl-1.
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Four types of media were used in the studies: 1. peat substrate, 2. mixture of sphagnum peat - pine bark - perlite - coarse sand and clay (1:1:1:1:1), 3. peat mixed with perlite (1:1), 4. peat mixed with sand (1:1). For preparation of each type medium, 1 g multi-component Azofoska mixture per 1 dm³ was used. Four-centimeter fragments of rhizome were rooted in pots of 14 cm diameter filled with 4 types of media. The experiments were carried out in spring 1993, 1995 and 1997 using the Yellow King cultivar, and also in spring (May) and in summer (August) 1995 with the cultivars: Appelbloesem, Mona Lisa and Yellow King. In order to evaluate the effect of fertilizer level in the medium on rooting and growth of plants, in summer 1997 the rhizome fragments of Yellow King cultivar were inserted in peat substrate for preparation of which the multi-component Azofoska mixture was applied in following amounts: 0.5; 1.0; 1.5; and 2.0 g per 1 dm³. After three months, the rooting of plants and their growth were determined on the basis of length increment, fresh weight of rhizome, number of branching, general number of shoots and their fresh weight. The medium type affected the growth of plants obtained from rooting of Alstroemeria rhizome fragments. The young plants of best quality for plantation in a permanent place were obtained by rooting of the rhizome in peat substate prepared with addition 1-1.5 g/dm³ of the multi-component Azofoska mixture. The peat substrate ensured good rooting results also under conditions unfavourable for Alstroemeria growth, i.e. at high temperatures during summer.
Experimental infection of Alstroemeria seedlings with naturally infected lily ‘Casablanca’ with stunting and flower bud deficiency phytoplasma resulted 3–4 weeks after top grafting in chlorotic and/or necrotic stripes, whitening of the leaves, shoot necrosis and die back. Flower discoloration or malformation were not observed. Attempts to transmit phytoplasma from naturally infected lily and experimentally infected Alstroemeria to Catharanthus roseus by top grafting resulted in stunted growth, dull yellowing and malformation of the leaves in 4–6 weeks. Some plants were temporary entirely vegetative and did not produce flowers. The periwinkle plants that were bridged by Cuscuta odorata from the diseased lilies and Alstroemerias showed similar symptoms as top-grafted ones. With the universal primer pairs rU3/fU5 specific PCR product with expected length ∼900 was amplified from samples collected from lilies with severe symptoms and top grafted test plants. All PCR products used for RFLP analysis after digestion with Alu I showed the same restriction profiles. Position of three obtained bands corresponded to the lengths of the DNA fragments of American aster yellows (AAY) phytoplasma group.
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