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Further information to the acclimatization of "in vitro" plants
54-58.Views:155The experiment was carried out with in vitro propagated 'MM 106' apple-rootstock plantlets. The transpiration of the plantlets was examined, and the changes followed by SEM analysis.
Data about the transpiration intensity of the acclimatized plants, of its value under different conditions of relative humidity and influenced by the existence of roots, as well as by the degree of acclimatization are presented.
Leaves were also examined and it was found, that stomata of in vitro developed leaves closed slowly, and the number of stomata of newly developed leaves decreased.
It is also shown, that in vitro propagated roots, generally, lose their hairs during acclimatization, but these roots are all the same important, as new roots of full value develop out of them.
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Application of the Jerusalem artichoke (Helianthus tuberosus L.), as a plant origin medium additive, during the micropropogation of Ada keiliana
61-64.Views:190A procedure for in vitro propagation of Ada keiliana seedlings are suited for acclimatization, was worked out. M medium was supplemented, with Jerusalem artichoke, as plant origin complex additive. The apply of JAD (1,5g/flask) gave the best response, considering the shoot (29 mm), and the root development (24,9) mm) too. The plantlets with satisfying growth (25-30 mm, 4-5 roots) were transferred in small pine bark: Novobalt peat: coconut fibres: perlit (2:3:1:1) mix, among greenhouse circumstances.
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The effect of different biostimulators on morphological and biochemical parameters of micropropagated Hosta ’Gold Drop’
22-29.Views:249During in vitro multiplication of Hosta ‘Gold Drop’, 20 g l-1 sucrose, 5.5 g l-1 agar and 4 concentrations (0.1-0.8 ml l-1) of Ferbanat L, Kelpak, Pentakeep-V were added to half-strength Murashige and Skoog (MS) basal medium. As compared to the control and other biostimulators, plants with lower peroxidase activity, larger fresh weight, more, longer shoots and roots, larger leaves were developed on medium containing Kelpak. The best concentration was 0.4 ml l-1 for in vitro rooting, shoot formation, plant weight and ex vitro chlorophyll, carotenoid level, peroxidase activity. Pentakeep was the less efficient biostimulator, increasing of its concentration mostly decreased root and shoot values (furthermore, abnormal callus formation was observed, as non-wanted effect), chlorophyll content and sizes (length, width) of leaves, not only during in vitro propagation but also (as after-effect) acclimatization because of the high mortality and weakly developed survivor plants.
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Results in the vitro propagation acclimatization of Pontederia lanceolata
47-49.Views:116This paper gives an outline of micropropagation of Pontederia lanceolata. Pontederia l. is a widely used aquatic plant, therefore there is an increasing demand for them, which can be satisfied only by in vitro culture. Research was carried out to find the best nutrient media conditions for micropropagation and acclimatisation of Pontederia lanceolata.
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Morphological, physiological features and differences of Vriesea splendens ’Fire’ plants during in vitro multiplication and rooting
Views:120During in vitro multiplication and rooting of Vriesea splendens ’Fire’, 0.1, 0.2, 0.4 and 0.8 mg l-1 benzyladenine (BAP), benzyladenine-riboside (BAPR), kinetin (KIN), meta-topoline (MT), indole-butyric acid (IBA) and naphthalene-acetic acid (NAA) were added to basal Murashige and Skoog (1962) MS medium. As compared to the hormone-free control, plants developed significantly more shoots on medium supplemented with almost all cytokinins (excepting KIN), especially BAP resulted the highest multiplication up to almost 26 shoots. Enhancement of cytokinin concentrations increased shoot number (and in case of BAP, peroxidase activity) but decreased plant height and rooting parameters. Regarding root production, both auxins were definitely beneficial (0.2 mg l-1 NAA resulted more than 7.5 roots and higher auxin concentrations efficiently stimulate root elongation); however, KIN had similar effects. After a three-month duration time of acclimatization, we observed that plants which were previously cultured on medium containing certain cytokinins (KIN in all doses and 0.1 mg l-1 MT) or both auxins had greater survival, moreover, as negative after-effect, higher cytokinin concentrations reduced the number of survived specimens.
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Ultrastructural and biochemical aspects of normal and hyperhydric eucalypt
61-69.Views:239Hyperhydricity was observed throughout in vitro multiplication phase of a Eucalyptus grandis clone. Ultrastructural approach of tissue and cell differentiation, izoenzyme patterns, binding protein (BiP) expression, and pigment content were performed. Hyperhydric tissues showed a reduction in cell wall deposition, reduction of membranous organelles, higher cell vacuolation, and more intercellular spaces than its normal counterpart. Additionally, several vesicles were present in hyperhydric cells suggesting the occurrence of organelle autophagy by autophagic vacuole. Lower pigment content, intercellular spaces on the epidermis and the induction of a molecular chaperone (BiP) were observed in hyperhydric phenotype. Evidences of schizolysigenous process of intercellular space formation are compatible with a stress condition. Although plastoglobulli were observed in normal and hyperhydric chloroplasts, they were more evident in the normal ones. Abnormal stomata also reflected a disruptive situation and morphogenesis disturbances which would difficult plant acclimatization. Further observation of the epidermis ultrastructure allows us to conclude that the presence of intercellular spaces on its surface may be constraining the recovery and development of hyperhydric plants. Similarly to BiP, other proteins such as esterase (EST), acid phosphatase (ACP), malate dehydrogenase (MDH) and peroxidase (PDX) are possible to be used as stress markers in in vitro conditions. Our results confirm earlier findings about negative effects of hyperhydricity on in vitro plant morphogenesis and ultrastructure, which in eucalypt is associated with a stressful condition contributing to lower propagation ratios.
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In vitro multiplication and hardening of grapevine plants in aeriated media
15-18.Views:215In vitro cultures have widely been used in horticulture for rapid multiplication of new varieties and clones as well as to produce pathogen-free stock material. To improve efficient hardening and transfer in vitro grown grapevine plants were multiplied by cutting them into single-node internodes with the whole leaf. Microcuttings including the shoot tips were rooted in granulated perlite moisted with tapwater under sterile conditions. After 2-3 weeks the rooted microcuttings were supplied by nutrients and hardened by gradual opening and finally by complete removal of the lids of jars or plastic boxes used for growth. Using this method microcuttings of Vitis vinifera cvs. „Chardonnay", „Cabernet franc", „Riesling" and „Sauvignon blanc" and the rootstock varieties Vitis riparia x Vitis cinerea cv. „Barrier" and Vitis berlandieri x Vitis rupestris cv. „Richter 110" formed new roots and shoots and 100% of the tested plants survived the acclimatization procedure. Similar results were obtained when perlite was replaced with rockwool-, or pit-pot blocks. This method may highly increase the efficiency of producing pathogen-free propagating material and new transgenic lines.
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Nutrition of the micropropagated fruit trees in vitro and ex vitro
43-46.Views:147Some experience or details are introduced in connection with the nutrient uptake of micropropagated fruit trees in the different phase of the in vitro or ex vitro development. It can be stated, that the plants during the micropropagation procedure are overfed. Special careful nutrient supply is necessary during the acclimatization.