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In vitro regeneration from cotyledons of watermelon
96-98.Views:285Cotyledonary segments of five different genotypes of watermelon were used to induce organogenesis. Five different hormone combinations were applied to enhance the induction of shoot formation on the surface of the segments. The phases of organogenesis were followed with light and scanning electron microscope. Shoots were obtained after four weeks, then the shoots were transferred to hormone free medium for root induction.
This method of regeneration can be applied in transformation experiments. GUS histochemical assay was made to check the expected success of using Agrobacterium for the transformation.
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Obtention of new ornamental leaf variants of giant reed (Arundo donax L.) originated from somatic embryogenesis and their photosynthetic parameters
18-24.Views:1043Giant reed (Arundo donax L.) is a perennial rhizomatous herbaceous plant, it has been widespread all over the world by human activities. It is a 2-8 meter high, polyploid, sterile species (not produce viable seeds), it can be propagated only by vegetative methods (rhizomes, stem cuttings, in vitro biotechnological methods). It has considered promising dedicated energy crop thanks to high biomass production (20-40 dry tons per hectare depending on microclimate), adaptability of different kind of soils and environment and low energy input required for its cultivation. It has been utilized for energetic purposes, biogas/bioethanol production, cellulose/paper production and ornamental purposes. The objectives of the present study were to determine morphological properties of new ornamental giant reed leaf variants originated from somatic embryogenesis and to evaluate their photosynthetic pigment content and photosynthetic activities. The most typical changes was the appearance of different colour (white, yellow, light green) longitudinal stripes on leaves (also on petiole and on leaf blade). It was significant differences (p<0.05) between green and leaf variants in case of photosynthetic pigments content and photosynthetic activity (Fv/Fm value). There was no detectable chlorophyll a or b content in the white bands of leaves and albino shoots. Total chlorophyll content of the white striped leaf variety was more than twice than the light green leaves. Photosynthetic activity and content of photosynthetic pigments also confirm and determine the morphological characteristics and growth habit of leaf variants.
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Bean tissue culture and genetic transformation with Agrobacterium
32-35.Views:330In this paper we report the establishment methods of a rapidly growing callus culture of Phaseolus vulgaris bean as well as the conditions required for a high level of transient gene expression using Agrobacterium-mediated transformation. A vector is containing both the lindan-resistance gene as a selectable marker, and GUS gene as a screenable marker. By using hypocotyl explant and vertical culture on B5 medium supplemented with 1 mg/1 kinetin- and 2,4-D 2 mg/1 and subcultured every 3-4 weeks, we can recommend to get a good and much callus from bean. This will help in introducing foreign DNA into callus cells. One strain of Agrobacterium carrying plasmid as vector for introducing foreign DNA into plant cells was used. At different concentrations of lindan; 3, 4 and 4.5 mg/I, the transformed Maxidor callus survived and grew over a period of 6 month and subcultured every 3-4 weeks, but the control callus died. Callus were assayed for GUS activity to confirm the expression of the GUS gene using the histochemical assay test. The GUS gene was also correctly expressed in callus cultures grown on 4mg/I lindan-selected medium, the typical blue colour in the histochemical assay using the X-gluc as substrate. But the control, non-transformed callus was not able to grow in the presence of lindan, neither showed a positive reaction in the in vitro assays.
