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Vegetative propagation of peony is usually carried out by grafting onto herbaceous peony roots (tree peony) or by dividing tuberous root clumps (herbaceous peony). However, conventional propagation is time-consuming and too slow for a large production and therefore in vitro methods have been developed. The regeneration ability of different organs of herbaceous and tree peony has been reported in this review. The role of exogenous and endogenous growth regulators in the differentiation and growth of shoots, roots and somatic embryos is presented. The problem of dormancy in the somatic embryos and in the shoots or plants propagated in vitro are also discussed.
The influence of temperature, daylength and sucrose concentration on the growth and development of Alstroemeria 'Zebra' in vitro was investigated. Only slightly more lateral rhizomes were formed at 25"C in comparison with 17°C on the medium with BA. Presence of BA in the medium strongly increased number of upright growing shoots and more shoots were formed in temperature 25°C than in temperature 17°C. Rhizome multiplication and formation of upright growing shoots were not significantly influenced by daylength. Sucrose was required for the formation of lateral rhizomes, upright growing shoots and roots. The highest number of lateral rhizomes was observed on the medium containing 60 and 80 gl-1 sucrose and BA. Presence of BA in the medium with different sucrose concentrations markedly influenced the formation of upright growing shoots; the highest number of shoots was found on the explants cultured on the media with 20 and 30 gl-1 sucrose. Low and high concentrations of sucrose inhibited upright growing shoots formation and their elongation. Rhizogenesis was not observed on rhizomes growing on the medium without sucrose and with 10 gl-1 sucrose, either in the presence or absence of NAA. The number of roots per plantlet on the medium with NAA increased with increasing sucrose levels. The highest sucrose concentrations slightly inhibited roots formation.
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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The genus Helleborus belongs to the family Ranunculaceae and comprises about 22 species, which are distributed over different parts of Europe and West Asia. In Poland, only H. purpurascens is native and it occurs in the Western Bieszczady Mountains. Hellebores are popular as ornamental cut flowers and medicinal plants in Europe and the USA. Conventional propagation by seeds or division has a low multiplication rate and is time-consuming. Vegetative propagation is necessary to maintain the desirable characteristic of a particular hellebore cultivar. Although some research on tissue culture of hellebores has been published, effective commercial micropropagation of these species has not been attained because cultivation in vitro is still very difficult. This review presents the progress in Helleborus species propagation in vitro for its commercial production. The efficacy of hellebore micropropagation (initiation and stabilization of culture, multiplication and rooting in vitro and acclimatization ex vitro) has been influenced by several factors, such as: type of initial explants, genotype, growth regulators, and environmental factors (temperature, sucrose, nitrogen salts, phosphorus). The genotype-dependence of multiplication and rooting in vitro, and acclimatization ex vitro of some Helleborus species has been presented.
Herbaceous peony plants successfully propagated in vitro do not survive the transfer to the ex vitro environment. For other species, storage organ formation in vitro can limit the loss of plants during acclimatization. In the natural conditions, the renewal buds for the following year originate on the underground crown (metamor­phosed underground shoot, rhizome) of herbaceous peony. A perennial crown and roots serve for the accumulation of the storage products and plant renewal. The aim of the experiment was to investigate the influence of glucose (30, 60 90 g l-1) and growth regulators (kinetin, IBA, GA3) on the shoot, renewal bud, root growth and development of Paeonia lactiflora 'Jadwiga' in vitro. Excision of all leaves from isolated explants inhibited production of new shoots and leaves, and evidently induced formation of renewal buds. Increasing the glucose supply, espe­cially in the absence of growth regulators, decreased production of shoots and out­growth of leaves. The stronger inhibition of shoot growth by glucose was observed on the explants without leaves. By contrast, the beneficial effect of glucose on renewal bud formation was observed. A single supply of kinetin, IBA or GA3 stimulated shoots and leaf growth and inhibited renewal bud formation, on the explants isolated with leaves. Interaction of kinetin, GA3 and IBA (added together) and the highest glucose level enhanced the growth of shoots on the explants containing leaves, and increased the number of renewal buds, on the explants without leaves. Increasing glucose level enhanced the number of roots in the absence of growth regulators on the explants containing leaves. The supply of IBA in the medium containing 30 g l-1 glucose, stimulated the root production on the explants without leaves. The addition of GA3 or kinetin (singly or simultaneously with IBA) to the medium with different concentrations of glucose, strongly inhibited rooting. Vol. 18(2) 2010: 309-320 The results presented here, show that a high level of glucose and exogenous growth regulators (kinetin, GA3, IBA) together stimulate shoot and renewal bud for­mation but the way of organogenesis depends on the presence or absence of leaves. The interaction between auxin (exogenous or endogenous) and glucose regulate root formation on the peony shoots but the final effect depends on the type of explants (with or without leaves). It is possible that leaves have a very important hormonal factors, which stimulate shoot growth or rooting and inhibit renewal bud formation.
The influence of sucrose (5, 10, 20, 30 g l-1), nitrogen salts - KNO3, NH4NO3 (25%, 50%, 100% in relation to the MS medium) and temperature (15 °C, 20 °C) on the growth of the main shoot and the activation and development of axillary buds in Syringa vulgaris in vitro was investigated. Different ratios of sucrose/nitrogen salts in the MS medium had a limited effect on the length of the main shoot of lilac plantlets. Also, the concentration of sucrose and nitrogen salts in the medium did not signifi­cantly affect the formation of nodes on the main or axillary shoots. The outgrowth of axillary shoots depended on the sucrose and nitrogen salts concentrations and tem­perature. Among the various sucrose/nitrogen salts relations, the highest number of axillary shoots (4.2) was found in the plantlets growing at a temperature of 20 °C, on a medium with a low level of sucrose (5 g l-1) and 100% strength of KNO3 and NH4NO3. Increased levels of sucrose in the medium significantly reduced the devel­opment of axillary buds in lilac plantlets growing at either temperature. By contrast, high levels of sucrose increased the fresh weight of lilac shoots. Different levels of nitrogen salts in the medium containing the same level of sucrose had no significant effect on the fresh weight of lilac shoots. On the other hand, at all levels of sucrose, the increased strength of nitrogen salts in the culture medium significantly enhanced the emergence and growth of axillary shoots. Increased strength of nitrogen salts in the medium appeared to counteract, at least partially, the inhibitory effect of a high sucrose level on the growth of axillary buds in Syringa vulgaris. There was clearly an interaction between the levels of sucrose and nitrogen salts such that a medium with a low sucrose to nitrogen ratio promoted axillary branching, whereas a medium with a high sucrose to nitrogen ratio inhibited the growth of axillary shoots. The different ratios of sucrose/nitrogen salts in the MS medium and the temperature affected the morphology of lilac plantlets. Increased supply of sucrose strongly stimulated leafsurface area, but the levels of nitrogen salts had a limited effect on leaf size. The plant- lets cultured at a temperature of 15 °C had bigger leaves than the plantlets at 20 °C. Low-sucrose treatments, irrespective of the level of nitrogen salts, induced a compact and branched habit of shoots and inhibited root formation. Increasing sucrose content in the medium resulted in a spontaneous formation of roots on the plantlets cultured in the presence of low levels of nitrogen salts.
The effect of white, blue, green, red and UV + white light on the growth and development of shoots and roots of Gerberu jumesonii cv. Queen Rebecca in relation to the presence of kinetin or IAA were investigated. The highest number of axillary shoots was obtained in red and green light on the medium with 5 mg l-1 kinetin. Also, green and red light markedly increased the number of leaves developed on the plantlets on the medium supplemented with kinetin. Light quality and IAA added to culture medium variously affected the development of root system: roots were regenerated under all light treatments, higher root number was recorded under red light when 5 mg T' IAA was added to the media, the shortest roots were found in red light on the medium supplemented with IAA. The greatest fresh weight of shoots was found under white light on the medium with kinetin. Red light markedly decreased shoot fresh weight on hormone-free medium. Blue and white light caused increase in fresh weight of roots.
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