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Comparative investigations on protoplast culture of some Brazilian and Hungarian sweet pepper cultivars and hybrids
39-45.Views:352Cotyledon protoplasts were isolated from 16-18-day-old in vitro grown seedlings of 9 Brazilian and 3 Hungarian pepper varieties and hybrids. Large numbers (average 9.59 X 106 protoplasts g 14 fresh weight) of highly viable (average 87.0%) protoplasts were released using a pectocellulolytic enzyme mixture. Protoplasts were cultured in K8p mediuni using an alginate disc embedding method. The osmotic pressure of the medium surrounding the alginate-embedded protoplasts was reduced by replenishing the liquid medium at K8p:K8 ratios of 1:0. 2:1, 1:1 in the first. second, and third week, respectively. Initial plating efficiency (IPE) average was 38.5% and after 21 days protoplasts reached microcolonies (15-20 cells) stages. Microcolonies were transferred after 3-4 weeks to a MS-based medium supplemented with 1.0 mg I-1 zeatin, 3.0% (w/v) sucrose, 0.24% (w/v) phytagel and pH 5.8, whereupon they formed callus. Final plating efficiency (FPE) average was 0.29% at a plating density of 1.0 x 105 protoplasts Protoplast-derived calli were cultured on a range of MS-based media supplemented with either BAP, IAA, TDZ; and zeatin. No morphogenic response was observed in any genotype investigated.
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Physiological and biochemical evolution of peach leaf buds during dormancy course under two contrasted temperature patterns
15-19.Views:218Budbreak anomalies in temperate fruit trees grown under mild conditions have often been described. However, only few authors approached the physiological evolution of leaf buds all along the dormancy period according to the temperature pattern. The aim of this study was to characterize the evolution of peach leaf bud dormancy through some physiological and biochemical parameters under temperate winter conditions and under total cold deprivation after the endodormancy onset. Two treatments were applied in peach trees cv. Redhaven: (i) Regular Chilling Amounts — RCA and (ii) Total Chilling Deprivation — TCD. Buds were sampled periodically from different parts of the stem (terminal, medium and basal ones). We recorded the evolution of: carbohydrate concentrations (glucose, fructose, sucrose, sorbitol and starch), respiration rate, water contents and energy metabolism (ATP and ADP ratio). The dynamics of these parameters were compared and correlated with dormancy evolution ("one node cuttings" test) and budbreak patterns in plank:. The endodormancy intensity of terminal buds was significantly lower than those of median and basal buds in early October. Under RCA treatment, this gradient faded and the bud endodormancy release was completed at the same time in all positions along the stem. Thereafter, the "cuttings" test indicated that terminal buds grew slightly faster than median and basal buds, and, consistently, budbreak in planta started with the terminals buds, followed by the medians and then by the basal ones. The carbohydrate contents showed a transitory change only when the buds began to grow after the endodormancy was released under RCA. Respiration, water content and ATP/ADP changed dynamics only under RCA and only after the end of the endodormancy (their respective changes were very parallel). The dynamics of none of the tested parameters could be related with the endodormancy dynamics, but respiration, water content and ATP/ADP could be consistent markers of the actual bud growth before bud break (in this respect, ATP/ADP could not show differences between the terminal and axillary buds while respiration and water content could).
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How to produce large sized microtubers of potato cv. Desiree
33-36.Views:340In vitro tuberization was induced on explants with different number of nodes layered on a medium with high sucrose (8%) content: 30, 15, 10, 7 and 6 explants per jar were cultured containing 1, 2, 3, 4 or 5 nodes, respectively. Microtubers developed were graded by their smallest diameter, and the number of tubers per jar, their size distribution, their fresh weight and the multiplication rate were recorded. The highest multiplication rate (1.98) was obtained for explants with 5 nodes. The size distribution of tubers was markedly affected by treatments. The majority of microtubers (49.4%) were 6-8 mm in the case of the smallest explants (with I node). When explants with 2 to 5 nodes were used, the most microtubers were 8-10 mm but with an increase of explant size, more and more microtubers were produced with larger diameter up to 16 mm and average fresh weight of tubers also increased with the increase of explant size. For the microtuber production of Desiree the use of explants with two nodes can be suggested because in this treatment the average fresh weight of microtubers was high enough (250 mg) and the number of large sized microtubers was very high (79% was larger than 6 mm and 53% was larger than 8 mm).
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Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)
15-12.Views:420In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants.
Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of 0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance .
