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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple
7-10.Views:311Quince (Cydonia oblongaMill.) is a minor fruit crop, which is primarily used for marmalade, jam and sauce.Very few quince cultivars are known all over the world and in many cases similar names are used for presumably different cultivars. The aim of the present study was to evaluate and characterize the genetic diversity of 36 quince cultivars and selections with SSR markers. Seven out of 8 SSR markers designed from apple sequences could successfully yield amplification also in quince cultivars. Number of alleles per locus ranged from 2 to 3 alleles. These allele numbers are quite low when compared to apple. It is supposed to be the consequence of a genetic bottleneck. In spite of the low allele number per locus, the 36 quince cultivars formed 30 different genotypes. The ratio of homozygosity was low, which might be coupled with the self-(in)compatibility phenotype of quinces. SSR markers proved unable to differentiate putatively closely related cultivars (e.g. ‘Bereczki’ and ‘Bereczki bôtermő’). In general, the level of polymorphism among the tested quince genotypes was much restricted due to the low allele number detected. However, it must be considered that the number of analysed SSR loci is not enough high to estimate the overall heterozygosity of the quince genome. Further experiments are needed and the SSR markers proved to be a reliable and useful tool for such analyses.
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Economic aspects of aged apple plantations in Hungary
13-16.Views:129Being competitive is a goal all over in the EU. Competition is free and getting closer among the competitors of the apple industry. In Hungary, one of the most important issues of apple industry has been what are the prospects for aged plantations, which account for 50% of crop land. Based on our results, 80% of those apple plantations on the down-grade yielding apple, the ones only for processing, do not meet the criteria of competitive production. For those plantations in better shape that yield 30-40 t/ha, of which at least 20-30% are for fresh production, it can he profitable. Because of the future trend in technology and economy, however, even the latter ones can not be considered being competitive in the long run.
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Nectar production and pollination in peach
123-126.Views:240Observations were made at two growing sites, Siófok and Szatymaz, in the years 1998 and 1999, on 16 peach varieties. The production of nectar was measured, the foraging behaviour of bees, fruit set and the effect of exclusion of bee visits for different periods were observed systematically.
Production of nectar confirmed earlier data, 9.09 mg per flower in average. There was large variation due to variety and date of observation. Bee visits were relatively abundant. At favourable weather, 1 to 30 visits/flower/day occurred in the average. Artificial hand pollination increased fruit set, substantially. Open pollination yielded superior fruit set than self pollination, without bees. Supplementary bee pollination can be regarded to be beneficial to peach production as well.
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Summer pruning of sweet cherry trees and an inquiry of winter frost damages
41-44.Views:246One of the most demanded research projects is the intensification of fruit production. The use of dwarfing stocks is a moderate solution as their effect is scarcely satisfactory. Climatic conditions of Hungary are continental in Eastern Europe, where Atlantic and Mediterranean effects are interacting with the continentals in a kind of basin with characters of its own. Capricious meteorological episodes are often disturbing the security of development and fruiting of trees:
• winter frosts are damaging the cambium and fruiting structures of trees
• late spring frosts destroy cambium and flowers
• early autumn frosts hurt the leaves
• excessive precipitation impairs the growing fruits
• drought periods during the summer caused water stress disturbing water husbandry.Vigorous stocks still prevail in the practice, and they ought to withstand challenges of weather hazards. The strong vigour of plants delaysthe process of senescence and the tendency of getting bald, and regeneration of plants is a sign of vitality. In present research, the trees have been trained on vigorous Prunus mahaleb stocks. Summer pruning was one of the important tools of intensive growing techniques. They were compared with traditional techniques and with plastic foil protected trees observing the vegetative as well as generative growth of them.
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Correlation of precipitation distribution and quality sweet cherry production
39-43.Views:196Sweet and sour cherry need 550–600 mm yearly precipitation. The critical period is 1–1.5 month after flowering, it is normally between 15.April – 15. June in Hungary. The rain induced fruit cracking is also a critical and costly problem for cherry growers. Fruits grown under arid conditions are less resistant against rainfall during harvest and up to 50–60% crack damage may occur. A computer program was developed to calculate the precipitation related production risks of sweet cherry. Focus of the research was Zala county. Spatial distribution of precipitation was compared in two directions (East and North of Zala county) based on the data of meteorological stations. The first results indicate that the developed method estimates the risks quite well, compared to the farm experiment results. The developed computer program can be parameterised according to the user’s requirements, this allows to take into account the real variety structure of a given orchard.
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Training systems of fruit trees in Hungary
123-127.Views:288Growing sites and soil conditions of Hungary warrant profitable production of several temperate fruits at elevated levels of quality. The climate of the Carpathian basin is a mixtures of three main climatic zones the prevalence of which may change seasonally: Atlantic, continental and Mediterranean, therefore, growing sites are rather various. Temperature minima of the winter and late spring frosts are the main elements of risk. In choice of the system of cultivation, regularity of yields and intensity are to be observed equally. Regular yields are particularly aimed in stone fruit cultures.
For apple and pear plantations of high density required for intense production are promoted favourably. Accessories of intense orchards (irrigation, supporting system, rootstocks, phytotechniques, etc.) are important. In peach and plum trees are trained to funnel-shape crowns, in general, intense-types are possible in plum, only. In apricots, a Hungarian speciality, the "umbrella" type of crown is applied, almost exclusively, according to Papp. In sweet and sour cherry, the harvest technique, manual or mechanised, according to the intended utilisation, are determining the form of training.
Red and black currants as small fruits are grown mostly as bushes or hedgerows without any supporting system designed to facilitate mechanical harvest. Raspberries and blackberries are grown as hedges on trellis. Gooseberry is a special case, being a low, thorny bush difficult to be picked. Thus grafted small trees are attached to a wire-trellis which helps to solve problems of plant protection too.
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Production data of wine grape gene bank (Vitis spp.) of University of Debrecen, east Hungary
32-36.Views:200National wine strategy of Hungary promotes the use of “flexible” grape cultivars. These enable producers’ best fit to wine market changes and expectations. This study is aimed to present data on the gene bank of the University of Debrecen, Hungary. Data were collected at a single site, between 2010 and 2018 in east Hungary lowland on acidic sandy soil, own rooted planting material. Our results showed that besides high yield and adequate cane production desired sugar content at convenient pH is to be awaited with moderate deviation between vintages. Presented concept demonstrate technological flexibility of cultivars by their average deviation from regression equation between increasing sugar and pH typical for the vintage composed of data of cultivars of the gene bank. Average positive deviation means higher sugar content at specific pH, thus higher sugar content at desired, conveniently low pH (3.0-3.2 pH).
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The morphology of stigmata in stone fruit species
45-48.Views:140The morphology of the stigma has been studied in 50 varieties belonging to 6 stone fruit species. The majority of samples had elliptical stigmata with some exceptions with circular form (Duane, Tuleu gras). The surface of the stigma is papillary, flattened in side view (sweet cherry) or bulging (apricot, peach). The suture of the stigma is clearly visible as a depression and the varieties may differ in this respect.
The size of the stigma depends highly from the season, although the varietal differences are maintained. The dimension of stigmatic surface is characteristic for the species expressed in square millimetres: sweet cherry 0.92 to 2.91; sour cherry 1.64 to 2.48; plum 0.83 to 1.80; oriental plum 0.53 to 1.15; apricot 0.57 to 1.69 mm2.
The size and morphology of the stigma changes according to varieties too, and it may used in description and identification of varieties. No correlation has been found between the size of stigma and the fertility relations (self-fertility or self-incompatibility) of the respective varieties.
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Some physical properties of apricots and testing apricot sorting machines
51-62.Views:221Examinations were carried out in the manipulating and packaging plant of Gyümölcsért Ltd, in Boldogkôváralja, to determine some physical properties of five apricot cultivars and to test the work quality of the apricot sorting machines. The size and the weight of the fruits were measured and two sorting machines were tested. The results are given in tables and diagrams. The conclusions are also summarised.
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Inhibition and recovery of germination and growing ability of seedlings under and after osmotic stress induced by polyethylene glycol in 8 pea genotypes
53-59.Views:123Germination and early seedling growth of eight pea genotypes were examined under and soon after different (5%, 10%, 15%, and 20%) PEG treatments. Seeds were germinated on PEG solution for 3 or 6 days and then further germinated and cultured on filter paper moistened by water for 3 and 6 days. The length and fresh weight of shoots and roots of seedlings were measured and used for evaluation of genotypes. Roots were less inhibited by osmotic stress than shoots similarly to other plant species. The variability among the genotypes was the greatest in the case of shoot growth at 5% PEG treatment and in the case of root growth at 15% PEG treatment. Results suggest that growing responses of genotypes after cessation of stress are more suitable for the evaluation of their osmotic tolerance, than their responses expressed during in vitro stress conditions. Genotypes with relatively high or low osmotic stress tolerance, respectively, could be distinguished with 6 days after recovery from 3-day-long 15% PEG treatment concerning the rate of shoot weight to root weight.
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Effects of aromatic cytokinins on structural characteristics of leaves and their post- effects on subsequent shoot regeneration from in vitro apple leaves of 'Royal Gala'
41-46.Views:158The effects of different aromatic cytokinins applied in different concentrations and combinations were investigated on the histology of in vitro apple leaves and their post-effects on subsequent shoot regeneration from these leaves were studied. Great differences in the anatomical structure of leaves could be detected originating from media containing different types and concentrations of aromatic cytokinins. The number of regenerated shoots per explant and the organogenetic index were used for the evaluation of the post-effect of aromatic cytokinins on shoot regeneration. The histological structure of leaves used for regeneration and their regeneration response showed a good correlation. When the pre-treatment caused a juvenile-like or less-differentiated structure, the number of regenerated shoots per explant increased and often vitrification also decreased and consequently the organogenetic index also increased. A strong interaction between cytokinin-content (type and concentration) of the pre-treatment medium and that of the regeneration medium could also be detected.
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Effects of activated charcoal on rooting of in vitro apple (Malus domestics Borkh.) shoots
98-101.Views:136Rooting of in vitro 'Royal Gala' shoots was studied under different conditions of root induction and root elongation phase. The rooting capacity was affected by both rooting phases. Very high rooting percentage could be reached with both liquid and solid root induction media. Raising the temperature from 22 °C to 26 °C during root induction phase increased the rooting percentage. Presence of activated charcoal in root elongation media can affect the number of roots per rooted shoots and can increase the rooting percentage, the length of roots and the rate of survival depending also on other conditions during rooting. Presence of NAA in root elongation media reduced the number and the length of roots considerably. Favourable effect of activated charcoal on rooting was mainly due to adsorption of NAA.
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Organoleptic evaluation of pear varieties grown at different sites
33-37.Views:226It was stated as a summary that a system of testing pear varieties on scale of 100 degrees, weighing 11 properties, gives a comprehensive information on qualities of pear varieties. The points of the form to be filled out, distinguishes the appreciation of accessory flavours and tastes from the main properties. Under the title of taste, sweet and acid taste are evaluated separately. The presently used system is a step forward as the composed characters do not influence the judgement of each other. In the representation of results, the 100 degree system was more efficacious than the usual 5-point system because there was no need of using fractions, and the graphic profile is more expressive, so the commodity is better visualised. The assembly of the jury was divided into two groups as below and above 35 year old persons. The two groups judged differently the acidity, sweetness and the thickness of the fruit peel. It was suggested that the choice of the produce should consider the differences in taste and give up the idea of being equally favourable for both groups. The samples of different growing sites revealed that the best quality is produced at Nagykanizsa, and this capacity would be able to supply the market also in the large super-markets and chain of department stores in Hungary.
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Effects of different cytokinins on the shoot regeneration from apple leaves of 'Royal Gala' and 'M.26'
69-75.Views:198The effects of different types of cytokinins on the shoot regeneration from leaf explants of apple scion 'Royal Gala' and apple rootstock 'M.26' were evaluated. Regeneration media contained either thidiazuron, or 6-benzylaminopurine, or meta-topolin, or zeatin, or kinetin, or their N9-ribosides, respectively, in the concentration range 0.5 to 8.0 mg 1-1. Effects of 'these cytokinins were evaluated on the percentage of regeneration (R%) and that of vitrification (V%) and on the number of regenerated shoots per explant (SN). Organogenetic index (0I) calculated from these data was used for the evaluation of efficacy of cytokinins. The course of shoot organogenesis also was followed using stereomicroscope. Types and concentrations of cytokinins applied in the regeneration media influenced each parameter significantly and the regeneration answer was strongly genotype-dependent. The best regeneration (SN: 11.08, 01: 7.5) was achieved in `Royal Gala' by using TDZ in concentration of 0.5 mg 1-1 (2.271,1M). There was a clear relationship between the effect on the regeneration efficacy and the chemical structure of cytokinins considering classical cytokinins, namely N9-ribosides applied in less concentration than nonribosides have the same or best regeneration effects except for 6-benzylaminopurine riboside. However, similar relationship could not be detected in the case of 'M.26'. SN was the highest (3.22) using 6.5 mg 1-1 (18.2011M) 6-benzylaminopurine riboside or 8.0 mg 1-1 (21.44 µM) meta-topolin riboside (3.18). SN was not significantly lower (3.12) by using 2.0 mg 1-1 (9.08 1M) TDZ, however, OI was about half as big (0.63 compared to 1.29 or 1.74 with 6-benzylaminopurine riboside or meta-topolin riboside, respectively). 'Royal Gala' had higher organogenetic ability, than `M.26': 3.5-fold higher shoot number per explant and more than 4-fold higher organogenetic index was reached with this cultivar than with 'M.26'. Moreover, the similar developmental stage of shoots could be observed 3-5 days earlier than in 'M.26' and if explants of 'Royal Gala' were further cultured with 3 weeks, SN increased from 11.08 to 24.42 on TDZ-containing regeneration medium, which might suggest higher organogenetic ability, too.
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Single and dual effects of different cytokinins on shoot multiplication of different apple scions
76-78.Views:163Shoot 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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Effects of integrated and ecological growing technologies on the growth and development of fruiting structures in new apple plantations
31-33.Views:166On the Horticultural Station of the Debrecen University, Pallag, two year old apple plantations of two varieties (‘Pinova’, ‘Golden Reinders’) have been studied, in autumn 2011. The growth of the cross section area of the trunk and the central axis were measured and compared to evaluate the effect of two alternative growing technologies: integrated and/or ecological. ‘Pinova’ proved to grow more intensely than ‘Golden Reinders’ independently from the growing technology. The effect of the technology, however, was expressed in ‘Pinova’, where the integrated technology produced more vigorous growth than the ecological one. The effect of varieties on growing intensity was more distinct than the effect of growing technologies. Under the ecological growing system, the difference between the varieties studied is more accentuated regarding growing intensity. By that reason, in ecological growing plantations of varieties with moderate vigour ought to be stimulated by all possible techniques (as pruning, nutrition etc.).
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Epidemiology of brown rot on two apple cultivars in an organic apple orchard
49-52.Views:164In a two-year-study, the temporal development of brown rot (Monilinia fructigena) on fruits was analysed in an organic apple orchard on an early (Prima) and one late (Idared) maturing cultivars at Debrecen-Józsa in Hungary. Out of five mathematical functions (linear, exponential, three-parameter logistic, Gompertz, Bertalanffy-Mitscherlich), the three-parameter logistic function gave the best fit to brown rot incidence of all cultivars in both years. Disease progress started at the end of June for cv. Prima and at the end of July for cv. Idared, then disease increased continuously from 6-8 weeks up to harvest in all cultivars. Descriptive disease variates derived from the three-parameter logistic function were used to analyse disease progress. These were: Yf, the final disease incidence; Y55, fruit incidence at day 55; Y95, fruit incidence at day 95; b and q, the relative and the absolute rate of disease progress, respectively; T1.5, the time when disease incidence reaches 1.5 %; M, the inflection point and AUDPC, area under disease progress curve. Descriptive disease variates were significantly different (P<0.05) for cv. Prima compared to cv. Idared, except for the relative and absolute rate of disease increase, b and q, respectively. The largest differences among cultivars were in the values of the AUDPC. Disease progress curves and descriptive disease variates were presented and the practical implications of the results were discussed.
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Effect of gelling agents on in vitro development of Amelanchier canadensis ‘Rainbow Pillar’
75-79.Views:246In vitro shoot multiplication responses of Amelanchier canadensis ‘Rainbow Pillar’ were studied on media solidifi ed with different gelling agents. The media were gelled either with 6.8 g l-1 fi brous agar-agar, or 50.0 g l-1 wheat starch, or 20.0 g l-1 Guar gum, or 15 g l-1 Isubgol or 50.0 g l-1 wheat starch mixed with 0.5 g l-1 Phytagel. Shoot cultures were grown for two months, thereafter the multiplication rates (number of newly developed shoots per explant) were counted and the length of shoots were measured. We found that the highest shoot multiplication of Amelanchier canadensis ‘Rainbow Pillar’ occurred on media gelled with Guar gum, while the longest shoots developed on media with Starch. About four-fold shoot number were obtained on media with Guar gum compared to the weakest results found on media gelled with Isubgol. Finally, considering all factors (shoot growth parameters, costs) the most economical gelling agent for Amelanchier canadensis ‘Rainbow Pillar’ was proved to be wheat starch among the tested alternatives which allows a 75.6% cost reduction.
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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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Seasonality of weather and phenology of reproductive organs of flower of sour cherry cultivars in Hungarian climatic conditions
75-80.Views:193Sour cherry (Prunus cerasus L.) is one of the most important fruit crop grown in Hungary, The flowering phenology and pollen shedding/stigma viability ratio, in an effort to elucidate the reproductive phenology of nine economically important sour cherry cultivars (Érdi bőtermő, Debreceni bőtermő, Csengődi, Kántorjánosi 3. Pándy 279. Úfehértói fürtös, Petri, Éva, and Oblacsinszka) all widespread in the Hungary, was studied at Újfehertó climatic condition. The main aims were to investigate how possible environmental cues influence timing and development of phenophases and whether different cultivars have different seasonal responses to these cues. Likely effects of climatic changes on phenological development patterns were also considered. Our results revealed important aspects of the reproductive biology of sour cherry flowers. The amplitude of the phenophase "beginning of blossoming" between cultivars did not exceed 6 days. There is maximum 3 days difference in blooming length of different direction in each cultivar. Pándy 279 showed high variability when the position of flowers changed on the tree shoots. Újfehértói fürtös was stable in all four directions of tree. The pollen shedding period and stigma viability ratio was well synchronous. Pollen shedding phenomenon occurred in the range limitation of secretary activity of stigmas in all five cultivars. Distribution of pollen shedding over the secretary activity of stigmas is almost good. In all five cultivars maximum pollen shedding occurred about the high temperature part of the day.
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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:208Rooting 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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Biological performance of grape varieties in Eastern Hungary
31-35.Views:201Eurasian and interspecific grape varieties and candidates were inspected for biological productivity in the variety collection of the University of Debrecen at the Horticultural Experimental Station in Pallag. In this paper, data are reported on yields (kg/stock), cane production (kg/ stock) and use-up index of wood yield of each cultivar in the experimental years 2011-2012. From the range of Eurasian (Vitis vinifera L.) white wine-grape qualified cultivars and candidates ’Ezerfürtû’, ’Generosa’, ’Jubileum 75’, ’Müller Thurgau’, ’Zöld veltelíni’,’B-11’, ’CSFT-92’, ’Pintes’, from the range of Eurasian (Vitis vinifera L.) red wine-grape qualified cultivars and candidates ’Kármin’, ’Pinot noir’, ’Alicante Bouschet’ and ’Rubintos’ were highlighted. Promising data of interspecific ’Aletta’, ’Csillám’, ’Kunleány’, ’Orpheus’, ’Refrén’, ’Taurus’, ’Viktória gyöngye’, ’Alföld 100’, ’Reform’, ’RF38/32’, ’Toldi’ white, and ’Dunagyöngye’ ’Pannon frankos’ red wine-grape cultivars and candidates were emphasized. Published data refer only to the biological performance of the cultivars. The thorough evaluation of varieties needs respect to data on resistance to fungal disease and climatic extremities, and other characteristics concerning production technology and oenological parameters.
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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:201Different 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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Effect of aromatic cytokinins and explant position on shoot multiplication of Asparagus offi cinalis L.
67-70.Views:241Asparagus offi cinalis has been widely studied, but little information is available about its in vitro response to exogenous cytokinin during shoot multiplication. To study the effects of different cytokinins on shoot multiplication of A. offi cinalis ‘Grolim’, in vitro culture was initiated from shoot segments cultured on media with Murashige and Skoog medium. Effects of different aromatic cytokinins (6-benzylaminopurine, 6-benzylaminopurine riboside and meta-topolin) applied in four concentrations (0.5, 1.0, 1.5, 2.0 mg/l) on shoot multiplication of ‘Grolim’ were tested. Effect of explant position (vertically or horizontally) on the shoot multiplication outcome was also studied. Both the length and the number of newly developed shoots were signifi cantly affected by explant position and cytokinin content of the medium. The highest numbers of shoots (4.9) were produced in the presence of 0.5 mg l-1 6-benzylaminopurine riboside when explants were paced horizontally onto the medium. Although the longest shoots (41.5 mm) developed on explants placed vertically onto medium supplemented with 2.0 mg l-1 meta-topolin, the lengths of shoots developed on medium with 0.5 mg l-1 6-benzylaminopurine riboside were also adequate in both explant position (29.5 and 33.6 mm placed horizontally and vertically, respectively).
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Model experiments for establishment of in vitro culture by micrografting in apple
47-49.Views:191Micrografting was used in our experiments for establishment of in vitro culture from one rootstock (`JTE-F') and three scion cultivars (`Remo', 'Rewena' and `Reanda') of apple. Shoot tips of these cultivars were harvested from field and grafted onto in vitro rootstock cultivars. Their survival and development were studied. 42-93% of shoot tips survived and developed further depending on cultivar. Impermanent browning of sticking agar-agar could be observed in 21-25% of the micrografts depending on cultivars but discolouration of agar-agar ceased within one week and did not cause any death of shoot tips. We used micrografting successfully for establishment of in vitro culture from cultivars, from which earlier with conventional methods the culture establishment was not possible because of hard tissue browning. However, further studies are necessary to ensure the survival and development of shoots after removing them from micrografts.