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Studies on the alkaloid production of genetically transformed and non-transformed cultures of Lobelia inflata L.
65-71.Views:134The investigations of the growth and alkaloid production of cell suspension-, callus-, organized- and hairy root cultures from Lobelia inflata L. proved that these cultures are able to synthesize the characteristic piperidine alkaloids of the intact plant. Alkaloid precursor amino acids (Phe, Lys) and plant growth regulators affect not only the growth and differentiation of tissue cultures but also their secondary metabolism. The synthetic regulator Sz/I I combined with Phe increased the total alkaloid content considerably in callus- and organized cultures; regulator Sz/28 especially increased the lobeline content (in organized cultures in response to Lys, in callus tissues as a result of Phe application). With the aim of optimizing growth and alkaloid production of the genetically transformed hairy root cultures of Lobelia inflata L. we studied the effect of some growth regulators (NAA, IAA, kinetin) and precursor amino acids (Lys, Phe). The kinetin had inhibiting effect on the growth and lobeline production of the hairy roots. The IAA and NAA increased the biomass formation and lobeline production. The highest lobeline level was detected in tissues cultivated on hormone-free medium containing Phe.
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Knot formation by Pseudomonas syringae subsp. savastanoi on the in vitro shoots of Sorbus redliana
59-62.Views:158Two strains of Pseudomonas syringae subsp. savastanoi were isolated from Forsythia sp. and Nerium oleander in Hungary in 1997. The effects of growth regulators produced by the bacteria were studied in different experiments. The strains were co-cultured with Sorbus redliana in vitro shoots without being in contact with the plant on solid media. Further culture filtrates in different concentrations were added to the culture medium. The growth regulators presented in the agar caused knot formation on the shoots and on the leaves in both kinds of culture. There were significant differences in the cultural and physiological characters, auxin and cytokinin activity of the strains of different origin.
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Development of in vitro propagation system for Atriplex halimus L.
123-129.Views:205Explants excised from adult shrubs were surface sterilized and cultured on Murashige and Skoog (MS) basal medium in the presence of plant growth regulators (PGRs) at different concentrations. A high multiplication rate of 7.2-fold was achieved every four weeks on MS medium supplemented with 4.44 μM BA, 0.49 μM IBA and 0.58 μM GA3. Rooting was achieved with 73% efficiency within 2-4 weeks on agar-gelled MS basal medium free of PGRs. Rooted plantlets were gradually acclimatized to field conditions over 5-6 weeks with 65% efficiency. For in vitro selection for salt tolerance, MS medium was supplemented with increasing concentrations of NaCl ranging between 25 and 1000 mM. This study has demonstrated that in vitro shoots could tolerate up to 600 mM NaCl with optimal growth at 200 mM, while higher concentrations of NaCl affected growth negatively. Growth and shoot number decreased with increasing NaCl concentration with all plantlets died at 1000 mM NaCl.
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Organogenesis in eggplant (Solanum melongena L. cv. Embu) as affected by antibiotics and growth regulators
76-82.Views:214The influence of antibiotics (cefotaxime, timentin, kanamycin and hygromycin) and growth regulators (indolacetic acid and 6-benzylaminopurine) was evaluated on eggplant organogenesis. Solanum melongena hypocotyl segments (6 to 10 mm length), taken from 16 to 20-days in vitro grown seedlings, were used as explants. The basic medium was composed by MS salts, Gamborg vitamins and 2% sucrose, solidified with agar 0.8% and pH adjusted to 5.7±0.2. Morphogenesis was impaired at 50 to 100 mg L-I kanamycin and 7.5 mg L-1 hygromycin. Both Timentin and cefotaxime reduced the frequency of regenerating explants meanwhile hyperhydricity was not affected. A decrease in root regeneration was observed with increasing cefotaxime concentrations, although, timentin had no effect on root regeneration, as compared to the control treatment. Interestingly, the number of adventitious roots was more noticeable at 0.25 mg L-I IAA plus 0.5 mg L-1 BAP. However, if just IAA was added led to higher number of regenerated roots compared to other treatments.
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The effects of growth regulators in proliferation of Sorbus redliana 'Burokvölgy'
77-83.Views:191The Hungarian cultivar Sorbus redliana 'Burokvölgy' was proliferated on Murashige and Skoog (MS, 1962) medium with half-strength macroelements and 100 mg/1 meso-inositol, 20 g/1 sucrose, 11 g/1 agar-agar. Different combinations of kinetin (KIN), metatopolin (mT), benzyladenine (BA), benzyladenine-ribosid (BAR) and indolebutiric acid (IBA) were tested, and pH was adjusted to 5.6 every case using KOH. The cultures were incubated at 20-24 °C in 8/16 hours dark/light photoperiod for 50-52 days. The main aim of our research was to find the optimal growth regulator and its optimum concentration. Purthermore, to determine the chlorophyll contents of the in vitro propagated plants' leaves. During the proliferation, the highest number of shoots were observed in the case of using BA + IBA, and on the medium containing 0.75 mg/I BA + 0.05 mg/1 IBA 8.93 shoots were found. The addition of KIN + IBA decreased the number of shoots and increased the sizes of leaves — the widest (11.2 mm) and longest (17.8 mm) leaves were obtained on the medium containing 1.00 mg/I KIN + 0.05 mg/1 IBA. The longest shoots (36.46 mm) were found in the case of applying 0.75 mg/1 BAR + 0.05 mg/I IBA. The BA + KIN + IBA combination resulted the shortest shoots. Sometimes not only shoot regeneration but spontaneous rooting was observed during the multiplication. The highest chlorophyll content (1.569 mg/g total chlorophyll, 1.132 mg/g chlorophyll-a, 0.437 mg/g chlorophyll-b) was obtained in the presence of 1.0 mg/I KIN + 0.05 mg/1 IBA.
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Vegetative and micropropagation potential of Piper guineense (Schumach and Thonn)
29-36Views:106The continuous loss of forest plants due to deforestation, and the increasing demand for Piper guineense because of its medicinal and food value, has put a permanent pressure on its population in the wild where it is collected. A method for conservation and mass propagation is therefore required. This research was undertaken to determine the optimal concentration of auxin needed for vegetative propagation and to investigate the potential of Piper guineense for micropropagation. The auxin optimization study of vegetative propagation was based on the use of two-nodal stem cuttings treated with five different concentrations of indole-butyric acid (IBA). Growth parameters such as the number of sprouted, rooted and survived cuttings among others were determined. To investigate the potential of Piper guineense for micropropagation, nodal explants were subjected to different sterilizing treatments using ethanol, NaOCl, mancozeb, streptomycin and Plant Preservative Mixture (PPM). The effect of plant growth regulators (PGRs) was tested on sterilized nodal explants using full strength Murashige and Skoog (MS) hormone-free media alone as control and MS media supplemented with PGRs (BA, NAA and KIN) at different concentrations and combinations. Significant differences were observed across the treatments for all growth parameters measured. However, 2000 ppm IBA significantly (p<0.05) influenced sprouting and rooting of the stem cuttings. Piper guineense explants have deep tissue contaminants, which cannot be eradicated by surface sterilization alone except double sterilization using PPM. On control media, neither shoot nor root response was observed while the highest percentage of induced roots was obtained from explants cultured on MS +1 mg/L BA + 0.25 mg/L NAA. Shoot induction was only achieved when BA was used alone and when subcultured on media supplemented with NAA, which generated roots.
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Experimental approach in apple tree nutrition
9-13.Views:123Authors present synthesis of experimental work, performed in the last decades, for better understanding nutritional behaviour of apple trees and related problems in fruit quality. There were evidences supporting possible deteriorating role of potassium in feeble physiological status of apples, if applied in excess. More intensive studies proved that higher potassium uptake into leaves and fruits might be also the result of increased sink power of individual fruits. Nevertheless early senescence of apples during storage and also sensibility to bitter pit were successfully related to the increased sink power of fruits, casual relations in excessive NPK fertilization, although increase in sink power need further investigations. Impaired weather conditions during early development of fruits, hostile orchard practices in pruning, thinning, irrigation and also unskilled application of growth regulators may also contribute in the enhancement of sink power and in weakened physiological status of apple fruits.
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Influence of different growth regulators on the in vitro morphogenesis of an ornamental variety of carnation
55-57.Views:137Callus formation, as a prerequisite for the induction of somaclonal variability, was achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Regeneration of new plants from shoot apex meristems could be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. These in vitro treatments may contribute to a more efficient micropropagation of the Rimini variety of carnation.
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Production of transgenic carnation with a heterologous 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase bifunctional enzyme cDNA
75-79.Views:112Transgenic carnations were produced with a modified mammalian bifunctional enzyme cDNA coding 6-phosphofructo-2- kinaseffructose 2,6-bisphosphatase. Relative activity of this enzyme determines the fructose 2,6-bisphosphate (fru 2,6-P2) cytosolic concentration. This metabolite — as a signal molecule — is one of the carbohydrate metabolism regulators. The regenerated Dianthus chinensis and Dianthus caryophyllus shoots were selected on MS basal medium containing 150 mg/1 kanamycin. Transgene integration was proven by PCR analysis with cDNA specific primers followed by Southern hybridization of DNA isolated from selected green shoots, which survived on kanamycin containing medium, so 3 D. chinensis and 20 D. caryophyllus transgenic plants were produced. Transgene expression were examined by RT-PCR. Transformed and control plants were potted in glasshouse to evaluate the effect of modified fru 2,6-P2 on development, growth and carbohydrate metabolism.
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Investigation of the in vitro regeneration of mericlones in the caribe variety of carnation
87-89.Views:130In vitro culture conditions were experimented for the relatively sensitive, but very esthaetic "Caribe" variety of carnation with uniformly dark violet flowers. Regeneration of new plants from shoot apex meristems can be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. Callus formation as a prerequisite for the induction of somaclonal variability can be achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Acclimation of the obtained mericlones to the ex vitro conditions was also evaluated.
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Factors affecting rhizogenesis in vitro and acclimatisation of three cherry rootstocks
40-46.Views:127Factors affecting rhizogenesis in vitro and acclimatisation of three rootstocks of cherry, i.e. Mahaleb, Maxma-14 and Weiroot -10 were investigated.
Rooting was easily achieved within 2-4 weeks on MS-based liquid or agar-gelled media containing auxins IBA at conc. 0.49 or 2.45 pM or NAA at conc. of 0.49 pM. On liquid media with 2.45 pM IBA, a maximum rooting efficiency of 95-100% was obtained. However, high concentrations of auxin delayed the time of root initiation for 3-5 days.
Rooted plantlets were transplanted into pots with a mixture of 3:1 (v/v) peat:perlite and acclimatised gradually to field conditions with efficiency of 60%.