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The effects of growth regulators in proliferation of Sorbus redliana 'Burokvölgy'
77-83.Views:203The 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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Single and dual effects of different cytokinins on shoot multiplication of different apple scions
76-78.Views:178Shoot multiplication responses of three apple scions to different concentrations of BA and BAR as single source of cytokinins and in combination with two concentrations of KIN were studied. The effects of hormones depended on genotype, type and interactions of different cytokinins. Use of BAR significantly enhanced the shoot multiplication of cv. Jonagold (6.5 shoots per explant). The multiplication rate of cv. Jonagold could not be improved by using the combination of BAR and KIN. The best proliferation was achieved by 1.0 mg 1-1 BA combined with 1.0 mg 1-1 KIN of cv. Prima..(8.1) and of cv. Galaxy (10.4).The effect of 0.5 mg 1-1 BA along with 1.5 mg 1-1 KIN was similar on multiplication rate (10.9) of cv. Galaxy.
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In vitro shoot multiplication of apple: comparative response of three rootstocks to cytokinines and auxin
36-39.Views:237Shoot multiplication responses of rootstocks cvs. M.26, MM.106 and JTE-H to different concentration of BA, BAR and IBA in eight various combinations were tested on MS-medium. The effect of hormones depended on genotype, type of cytokinin and interaction of cytokinin and auxin. Shoot multiplication was significantly enhanced with the use of BAR as cytokinin. High multiplication rate could be achieved in cvs. M.26, MM.106 and JTE-H: 7.7, 6.9 and 9.9 shoots per explant, respectively.
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The effect of different cytokinins on chlorophyll content and morphological features of in vitro Nidularium ’Kertész Jubileum’
47-51.Views:237During in vitro multiplication of Nidularium ‘Kertész Jubileum’, 20 g/l sucrose, 5 g/l agar, 100 mg/l inositol, and different concentrations of benzyladenine (BA), benzyladenine-riboside (BAR), kinetin (KIN), meta-topolin (mT) were added to the MKC (Knudson, 1946) basal medium. Furthermore, 0.1 mg/l naphthaleneacetic acid was used to every medium. Number of shoots, length of leaves, number and length of roots, chlorophyll (a+b) content were examined and evaluated with Ropstat statistical software. As compared to the other cytokinin, significantly most shoots were obtained in the case of applying BA. Increasing of BA-concentration (as far as 2 mg/l) enhanced shoot number (from 10.92 to 19.26) but 4 mg/l BA resulted only 6.63 shoot. The less efficient cytokinin was KIN, in most cases no more than about 2 shoot was achieved. Regarding the length of leaves, the higher level of BA effected averagely the shorter leaves (from 24,46 to 7.31 mm). KIN effected significantly the longest leaves (43.4-61.29) in inverse proportion to the concentration. The same cytokinin resulted the most (and the longest) roots with the highest rooting percentages, but more KIN decreased the number and length of roots (from 7.95 to 4.4 and from 38.49 to 22.73 mm). There were no definite correlation between cytokinin concentration and chlorophyll (a+b) content, but the highest doses resulted decreasing (except of meta-topolin which leads to the lowest values). Summarizing, BAR effected the highest contents (mostly more than 1400 μg/g), particularly in the case of 1 mg/l (1807.3 μg/g).
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Genetic transformation of bean callus via Agrobacterium- mediated DNA transfer
49-53.Views:122Callus cultures were induced from hypocotyl of young bean seedlings. Callus developed and maintenaned on B5 medium supplemented with 2mg/1 2,4-D and 1 mg/1 kinetin. The results demonstrate that A. tumefacins-mediated transformation is a convenient method to obtain transient gene expression in callus of bean. The results have shown that the bean callus co-cultivated with A. tumefaciens can be transformed to get heibicide Finale (glufosinate-ammonium) resistant GUS positive tissues. Southern blot analysis of transformed calli showed integration of gusA marker gene carried by a binary vector. Transformed calli were selected on herbicide containing media. Data of molecular analysis (Southern blotting) confirmed the insertion of gusA gene in the genome of herbicide resistant calli with bar gene. There are three evidences that calli are stable transformants: (1) herbicide resistance, (2) GUS activity which is indicative since the coding region containing an intron, (3) the results of Southern hybridization technique.
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Influence of aromatic cytokinins on shoot multiplication and their after-effects on rooting of apple cv. Húsvéti rozmaring
84-87.Views:219Different aromatic cytokinins (BA, BAR, TOP and KIN) were tested alone or in combination for the shoot proliferation response of ‘Húsvéti rozmaring' apple scion. The best multiplication rate was achieved by dual cytokinin application (1 mg 1-1 BA + 1.5 mg 1-1 KIN). The rooting capacity was affected considerably by the position of shoots: transfer of the three-week-old shoots to the same or other proliferation medium in vertical position inhibited the following rooting totally. Post-effects of different cytokinins (BA and TOP) on subsequent rooting could be detected: BA increased the number of roots markedly, while TOP resulted in significantly longer roots.
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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:276In 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 . -
The Effects of Some Parameters on Agrobacterium-Mediated Transformation in Muskmelon
46-49.Views:178Some parameters involved in Agrobacterium-mediated transformation in muskmelon Hales best (HBS) were studied. Cotyledon explants excised from 3.5-day-old seedlings were co-cultivated with Agrobacterium tumefaciens harbouring binary vectors which contained GUS and BAR genes. After co-cultivation on a low pH medium, explants were transferred to selective medium, with higher pH, containing Claforan and Finale. The medium was changed every two weeks till shoots were induced. All shoots rooted on MS medium supplemented with 0.3 mg/L IBA. These parameters combined as a whole led to successful transformation. The expression of the introduced gene construct was confirmed by GUS staining of shoot segments.
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Hazardous element content and consumption risk of 9 apricot cultivars
61-65.Views:281The heavy metals pollution is one of the problems that arise due to the increased uses of fertilizers and other chemicals to meet the higher demands of food production for human consumption. In order to assess possible health risk of apricot (Prunus armeniaca L.) consumption, levels of Arsenic, Cadmium, Mercury and Lead were determined in fresh and dried samples of "Jumbo cot", "Tom cot", "Gold strike", "Gold bar", "Bergeron", "Bergarouge", "Sweet cot", "Yellow cot" and "Zebra" apricot cultivars. Wet digestion of samples with concentrate HNO3 – H2O2 digester mixture and inductively coupled plasma–atomic emission spectroscopy was used. Highest content of As, Cd, Hg and Pb among all cultivars, were 0.5, 0.04, 1.5 and 0.5mg/kg of dried apricot samples. Fresh fruit samples also contain 0.2, 0.016, 0.6 and 0.2 mg/kg of Arsenic, Cadmium, Mercury and Lead respectively. Daily intake of metals, hazard quotient and health risk index to reveal health risk possibility of dried and fresh fruits consumption were calculate and compared.