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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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The effect of timing and IBA treatments on the rooting of plum rootstock hardwood cuttings
7-10.Views:214In propagation of plum by hardwood cuttings, the success of rooting is affected by several factors. Many authors deal with the timing of cutting collections, others investigate the optimal extent of hormonal stimulation. However, there is no data, as yet, about the coherence between these two factors. The aim of this experiment was to find the most advantageous condition s, with regards to both timing and IBA doses, for rentable propagation of the examined varieties. Five varieties were studied: 'INRA Marianna GF 8- 1', 'Myrobalan B', 'MY-KL-A', 'INRA Saint Julien GF 655/2' and 'Feher besztercei'. Cuttings were collected on seven occasions, during late autumn and winter, and were treated with IBA solutions of different concentrations. The optimal dose of IBA was found to be dependent both on the variety and the actual date of cutting collection . Results are reported, along with suggestions for optimal doses and collection periods.
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Determination of auxine content of soft wood cutted `Marianna GF8/1' (Prunus cerasifera x P. munsoniana) by High Performance Liquid Chromatography during rooting period
67-70.Views:125The content of different auxins of soft-wood cutted plum rootstock 'Marianna GF8/1' (Prunus cerasifera x P. munsoniana) was determined during the rooting period. The level of auxin-concentration (exogenous and endogenous) of basic and intemodal part of cuttings was determined by WATERS HPLC equipment every 7 days during rooting period. The lengths soft wood cuttings were app. 30 cm long. The basal part of shoots were treated with 2000 pg/g concentrated indole-butiryc acid in talcum powder. After treatment the cuttings were placed in propagation green-house under intermittent mist. The plant hormones were extracted by methanol the solution was cleaned by paper-filter, and further cleaned by centrifuge. The effluent was examined by reversed phase High Performance Liquid Chromatography, with WATERS 2487 dual detector at 220 am on Symmetry C18 4,6x 150 column. Recovery and reproducibility assessment indicates good accuracy and acceptable relative standard deviation (RSD) 5%. Linear responses (r20.997) for calibration curve was obtained with IAA, IPA and IBA standard in range, with a limit of quantification of 0.15 g•m1-1. The concentration of IAA, IPA and IBA in the basal part of cuttings were measured, during the rooting period. We proved the external IBA was taken up by the plants. In the plants were found the IBA, and the IAA concentration of IBA treated cuttings was higher, than the untreated one.
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Post-effects of cytokinins and auxin levels of proliferation media on rooting ability of in vitro apple shoots (Malus domestica Borkh.) 'Red Fuji'
26-29.Views:206Rooting ability of in vitro apple shoots of 'Red Fuji' grown on proliferation media with different hormone content were tested at three IBA levels in root induction media. Rooting percentage could be slightly increased with an increase in IBA concentration in proliferation media. The highest IBA concentration (3.0 mg 1-1) in root induction media showed strong inhibitory effect on rooting capacity of in vitro shoots. The highest rooting percentage (95%) could be achieved by shoots grown on proliferation media containing TOP or BA+KIN as cytokinins before rooting.
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Effects of indole-3-butyric acid levels and activated charcoal on rooting of in vitro shoots of apple rootstocks
25-28.Views:249Rooting responses of rootstocks cvs. JTE-F1, M. 26 and MM. 106 were studied to different concentration of IBA in root induction media and to presence of activated charcoal in root elongation media. High rooting rate (>90%) could be achieved in cvs. JTE-H and M. 26, while cv. MM. 106 showed weak rooting ability at each IBA level tested. Increasing IBA content depressed the rooting only in cv. M. 26. Presence of activated charcoal decreased considerable the rooting rate in cv. M. 26 and decreased the number of roots in cvs. JTE-H and M. 26. These cultivars developed longer roots on media containing activated charcoal, while cv. MM. 106 did not showed any reaction for it.
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Propagation of plum rootstocks by hardwood cuttings
23-28.Views:125Vegetative propagation by hardwood cuttings is a very simple and cheap method for production of plum rootstocks. The aim of this study was to examine if this propagation technique is suitable for practice of three plum rootstocks and find the time or period when the percent of rooted cuttings is maximal. Based on our results, hardwood cuttings of the rootstocks tested have the rooting potential acceptable for practice, however, for Fehér besztercei in the previous literature leafy cuttings are recommended. Fehér besztercei reached 74.0% rooting, cuttings of Sainte Julien GF 655/2 rooted in as high percentage as 78.3%, and Marianna GF 8-1 had 88.3% rooted cuttings. Rooting potential of hardwood cuttings depends on more factors, one of them can be their dissimilar sensitivity for the diverse environmental circumstances at the different propagating dates, affecting through the internal biochemical changes that can be in relation with the differences in their dormancy.
The cuttings of Marianna GF 8-1 take root easily, but in some years the conditions were less favourable for reaching maximal rooting. For taking cuttings the period from the beginning of October until December was optimal. For Fehér besztercei the optimal date of cutting collection was around the end of October, but in some years the rooting in the middle of December was also high. Sainte Julien GF 655/2 definitely rooted best in October. The treatments with different IBA concentrations in two years affected differently the rooting percentage. The rooting of Marianna GF 8-1 and Sainte Julien GF 655/2 is barely influenced by the different hormone dose in both years. Hardwood cuttings of Fehér besztercei rooted definitely better when treated with 2000 ppm IBA in comparison to untreated ones, while in 2001-2002 there was no difference between 2000 and 4000 ppm.
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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%.
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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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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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Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)
15-12.Views:266In 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 . -
In vitro rooting and anatomical study of leaves and roots of in vitro and ex vitro plants of Prunus x davidopersica 'Piroska'
42-46.Views:160The process of in vitro rooting and the anatomical characters of in vitro and ex vitro leaves and roots of Prunus x davidopersica 'Piroska' were studied. Best rooting percentage (50%) and highest root number (5.0) was achieved in spring on a medium containing 0.1 mg/I NAA + 30 g/1 glucose. At the end of rooting the parenchyma of the in vitro leaves was more loose and spongy, than during the proliferation period. In the first newly developed leaf of an acclimatised plant, the parenchyma was much more developed, contained less row of cells and less air space too, compared to the leaves developed in the field. The in vitro developed root had a broad cortex and narrow vascular cylinder with less developed xylem elements, but at the end of the acclimatisation the vascular system became dominant in the root.
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In vitro shoot multiplication of apple: comparative response of three rootstocks to cytokinines and auxin
36-39.Views:206Shoot 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 Effects of Some Parameters on Agrobacterium-Mediated Transformation in Muskmelon
46-49.Views:170Some 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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Morphological, physiological features and differences of Vriesea splendens ’Fire’ plants during in vitro multiplication and rooting
Views:114During 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.