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Effects of different cytokinins on the shoot regeneration from apple leaves of 'Royal Gala' and 'M.26'
69-75.Views:211The 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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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:167The 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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A study of processes active in regeneration of different sweet cherry varieties
55-57.Views:215Cherries belong to the group of fruit trees, which require little attention in pruning. The process of getting bald branches is less accentuated, which is associated with the light requirement of organs changing along with the aging process. The growing habits of the varieties (length and number of growing shoots, etc.) is closely related with the productivity of fruiting parts. The ability of regeneration and the functional effectiveness of fruiting structures are expressed by the distribution according to their age. The differences registered justify the necessity to develop pruning policies for each particular type of variety. For that purpose, we have to examine the age dependency of the processes related to regeneration, which may serve as starting point of a pruning strategy. Growth of some parts should be stimulated in order to develop or regenerate fruiting parts. Some branches need to be rejuvenated, partially, as an antidote to progressive senescence. Every intervention is aimed to strengthen the development of the most productive constructions. Reactions induced by rejuvenation should be expressed in fruiting intensity. A pruning program adapted to the particular variety as well as to the actual phase of a rotating pruning system should anticipate results.
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Study of different factors of grapevine regeneration systems and genetic transformation
33-36.Views:212The most limitating factor for successful transformation is the absence of high-yielding regeneration protocols. However, the anther-derived embryogenic culture is an optimal technique for genetic transformation and it has been widely applied in many important cultivars, but the necessity of further development of regeneration systems has been proved. We attempted to produce somatic embryos on a wide range of genotypes from various tissues; leaves, petioles, stem segments. We started the examination of grapevine regeneration via organogenesis, succeeded in inducing shoot from the meristematic tissue of the base of bud by testing induction medium contained different concentrations of two types of hormones. To optimize the conditions of the Agrobacterium-mediated transformation, we studied the effectiveness of different Agrobacterium-treatments, the use of antioxidants and the sufficient quantity of kanamycin for selection of transformed cells.
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Yielding capacity and regeneration of different ages fruit-bearing shoots in sour cherry
55-57.Views:178The yielding potential and regeneration (i.e. germination of hidden buds) after pruning of the fruit-bearing shoots of different ages were compared in sour cherry cultivars pruned to spindle shape. The examinations were carried out in order to determine the optimal pruning strategy resulting in the highest long-term yielding of sour cherry cultivars with intensive crown shape (free and thin spindle shape). According to our results, there are great differences among the studied self-fertile sour cherry cultivars in yielding and regeneration, which are worthwhile to be considered in determining the method and degree of pruning.
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Aminoglycoside antibiotics affect the in vitro morphogenic response of chrysanthemum and tobacco
93-104.Views:113Broadly the success of genetic transformation of plants requires non-chimeric selection of transformed tissues and its subsequent regeneration. With rare exceptions, most plant transformation protocols still heavily utilize antibiotics for the selection of transgenic cells containing an antibiotic-degrading selectable marker gene. The morphogenic capacity of in vitro chrysanthemum and tobacco stem and leaf explants change with the addition of aminoglycoside antibiotics (AAs). Of 6 antibiotics tested, phytotoxicity occurred at 10-25 and 50-100 pgml-I in chrysanthemum and tobacco explants, respectively, depending on the size of the explant and the timing of application. The presence of light or darkness also had a significant effect. The use of transverse thin cell layers (tTCLs) in conjunction with high initial AA selection levels supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow cytometric analyses demonstrate that regeneration can be predicted in both species, depending on the ploidy level of the callus. Endoreduplication was not observed in chrysanthemum, even at high AA levels, but occurred (8C or more) in tobacco callus, even at low AA concentrations (5-10 pgml-1). The higher the AA level, the greater the DNA degradation and the lower the 2C and 4C values.
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Attempting Regeneration from Cultured Cotyledons and Plant Regeneration from Cotyledonary Nodes in Common Bean (Phaseolus vulgaris L.)
57-60.Views:159Dry seeds from two cultivars of common bean (Phaseolus vulgaris L.) were germinated on sterile cotton and sterile deionized distilled water. Cotyledonary node tissue of seedlings were cultured on Murashige and Skoog(MS)-based media supplemented with different combination of N6-benzyl-aminopurine (BAP) and indole-3-acetic acid (IAA), and benzyladenine (BA) and a-naphthaleneacetic acid (NAA). The results revealed that the regeneration percent and the average number of buds and shoots per explant were influenced by the type of explants and exogeneously added hormones. Multiple shoot induction on dry bean cotyledonary node that contain 4-5 mm from cotyledons and hypocotyl on a medium containing full concentration of MS inorganic salts supplemented with 0.5mg/1 BA and 0.1mg/1 NAA was feasible and the method can be applied in transformation experiments.
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Investigation of the in vitro regeneration of mericlones in the caribe variety of carnation
87-89.Views:131In vitro culture conditions were experimented for the relatively sensitive, but very esthaetic "Caribe" variety of carnation with uniformly dark violet flowers. Regeneration of new plants from shoot apex meristems can be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. Callus formation as a prerequisite for the induction of somaclonal variability can be achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Acclimation of the obtained mericlones to the ex vitro conditions was also evaluated.
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Somatic embriogenesis and regeneration of Vitis sp.
41-45.Views:150Somatic embriogenesis and regeneration of Vitis sp.
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In vitro regeneration from cotyledons of watermelon
96-98.Views:166Cotyledonary 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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In vitro plant regeneration from immature embryo axis and cotyledons of common bean (Phaseolus vulgaris L.)
93-97.Views:128Phaseolus vulgaris L. is the most important economic species within the genus Phaseolus. It is grown in all parts of the world. Genetic improvement by conventional breeding has met considerable success, although production of hybrids between species within the genus has been limited due to sexual incompatibility. Recent advances in tissue culture have offered the opportunity to produce cultivars, which could not be obtained by conventional breeding methods. The use of tissue culture and genetic engineering is viewed as a logical approach to improve bean production. Gene transfer techniques will have a great impact on legumes. Although the concept of cell totipotency is widely proved, in vitro morphogenesis has not yet been achieved for a large number of cultivated beans. Regeneration protocols are strongly influenced by the genotype. In tissue and cell culture of beans, the factors controlling shoot morphogenesis and somatic embryogenesis are still unknown. The reported data suggest a possible way for future research.
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Shoot induction and plant regeneration from cotyledon segments of the muskmelon variety "hógolyó"
61-64.Views:143Cotyledonary segments of the casaba type muskmelon variety "Hógolyó" were used to induce organogenesis. Fifty different hormone combinations were applied to enhance the induction of shoot formation on the edge of the segments. The phases of organogenesis were followed with light- and scanning electron microscope. Shoot induction was achieved with high frequency. The shoots were transferred to hormone free media for root induction. The rooted plantlets were planted out to soil.
NAA was feasible and the method can be applied in transformation experiments.
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Organogenesis in eggplant (Solanum melongena L. cv. Embu) as affected by antibiotics and growth regulators
76-82.Views:229The influence of antibiotics (cefotaxime, timentin, kanamycin and hygromycin) and growth regulators (indolacetic acid and 6-benzylaminopurine) was evaluated on eggplant organogenesis. Solanum melongena hypocotyl segments (6 to 10 mm length), taken from 16 to 20-days in vitro grown seedlings, were used as explants. The basic medium was composed by MS salts, Gamborg vitamins and 2% sucrose, solidified with agar 0.8% and pH adjusted to 5.7±0.2. Morphogenesis was impaired at 50 to 100 mg L-I kanamycin and 7.5 mg L-1 hygromycin. Both Timentin and cefotaxime reduced the frequency of regenerating explants meanwhile hyperhydricity was not affected. A decrease in root regeneration was observed with increasing cefotaxime concentrations, although, timentin had no effect on root regeneration, as compared to the control treatment. Interestingly, the number of adventitious roots was more noticeable at 0.25 mg L-I IAA plus 0.5 mg L-1 BAP. However, if just IAA was added led to higher number of regenerated roots compared to other treatments.
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Preliminary results of renewal pruning an 18 years old sour cherry trees
41-42.Views:171Sour cherry is a light demanding fruit species. As most of the crop is developing from buds on one year old shoots and 2-yearold wood, the formation of bare wood can be seen apparently in a badly maintained canopy. The formation of „whip shoots” indicates the degree of bare wood. Unfortunately in many gardens and commercial orchards sour cherry trees resemble willow trees. Owners often decide to grub old orchards, although the useful cropping lifespan of the trees planted in a correct spacing can be 20-30 years on a move vigorous rootstock. Therefore renewal pruning can contribute to the full renewal of the orchard, but the length of its effect is depending on the growth characteristics and renewal capabilities of the cultivar. The growth characteristics of the cultivars are different. The regeneration capabilities of different aged wood are also different. These differences emphasize the need for cultivar specifi c pruning. Knowing the reactions to renewal pruning, we can ensure regular high crops with excellent quality by applying cultivar specifi c rotation pruning.
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Rate of scab infection and quality parameters of apple fruit in organic and integrated production systems
63-67.Views:190In this study, the rate of scab infection and quality parameters (content of vitamin-C, dry matter, sugar, total acids, specific weight, quality index and flesh firmness) of some apple cultivars are given, moreover, the relation between those characters was examined in two environmental systems (organic and integrated) in a rainy season. In the organic fruit production, scab infection of leaves was significantly higher than in the integrated system, in case of the same cultivars, which means that the rate of decrease in leaf area was heavier in the organic production. Cultivars did not differ statistically in parameters of inner content when the integrated system was compared to the organic one. Results did not show statistical difference between loss of leaf area caused by apple scab and the parameters of inner content, which is attributed to the regeneration of leaf area due to favourable weather conditions. Data on apple scab and inner content of the fruit justify essentially the environmental systems of apple production.
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Summer pruning of sweet cherry trees and an inquiry of winter frost damages
41-44.Views:265One 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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In vitro effect of different cytokinin types (BAP, TDZ) on two different Ocimum basilicum cultivars explants
15-20.Views:389Ocimum basilicum L. (sweet basil) is an economically and ethnobotanically important aromatic, medicinal, ornamental and culinary herb, with a very wide gene pool, that is sensitive to cold and prone to several plant pathogens that can demolish harvest and lessen yield. In this research, the effects of BAP (6-Benzylaminopurine) and TDZ (Thidiazuron) on different genotypes for in vitro cloning were determined, in order to provide a detailed protocol guide concerning Ocimum basilicum L. propagation. The results from the O. basilicum seed propagations revealed that the best condition for the secondary shoot growth is with 5.0 mg/l TDZ or 1.5 mg/l BAP on all types of explants except the root, the secondary root growth can be obtained on all types explant with any BAP concentration and all cytokinins can induce callus on all types of explants. On the whole, it shows that multiple secondary shoot induction and regeneration in Ocimum basilicum L. is regulated by appropriate cytokinin concentration.
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In vitro propagation of 'Echo' cultivars of Eustoma grandiflorum (Raf.) Shinn.
87-91.Views:145Eustoma grandiflorum (Raf.) Shinn. 'Echo' Fl cultivars ('Echo White', 'Echo Rose', 'Echo Blue', 'Echo Blue Picotee') were used and multiplication of shoots was evaluated on Murashige and Skoog (1962) basal medium with 11 g/1 agar-agar and 20 g/1 sucrose. To test the effect of BA different concentrations were added: 0.10, 0.25 mg/1 and a culture medium without BA. Differentiation of roots was examined on Jámbor-Benczúr and Marta (1990) basal medium with the same concentration of agar-agar and sucrose. To examine the effect on rooting, various concentrations of NAA were used: 0.5, 1.0, 2.0, 3.0 mg/l. The pH was adjusted to 5.6 in every case using KOH. We studied the after-effect of different concentrations of BA during the acclimatisation. During the multiplication, the cultivar 'Echo White' formed the most shoots and the smallest leaves on the medium with 0.10 mg/1 BA. Fortunately, in the case of this cultivar, the number of shoots was reduced and the length of leaves was increased succesfully on the medium without BA. The other three cultivars developed the longest leaves on the medium containing 0.10 mg/1 BA. Sometimes not only shoot regeneration but spontaneous rooting was observed during the multiplication. Examining the rooting, the highest percent of roots was found on the medium with 1.0 mg/1 NAA, and the cultivar 'Echo Rose' formed the most roots on this medium. Higher concentration (2.0 and 3.0 mg/1) of NAA already reduced the number of roots in all of the cultivars. During the acclimatisation, the percentage of survival was 76.3% and the tallest plants with the longest leaves were found on the multiplication medium with 0.25 mg/1 BA. 'Echo Blue Picotee' gave the best results with the tallest pieces and longest leaves on this medium.
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Tending operation models for black locust (Robinia pseudoacacia L.) stands growing on sandy soils in Hungary
55-57.Views:214A more intensive integrated research and development approach to the work carried out on the growth on sandy soils of stands of black locust (Robinia pseudoacacia L.) has been adopted in recent years, revealing several factors influencing stand growth. The fact that certain ecological factors influencing fundamentally the growth of trees have become unfavourable in Hungary in recent years has led to the more extensive use of black locust in the course of afforestation and forest regeneration schemes. The study presents a new, simplified tending operation model for black locust stands and age, growing space and target diameter models suitable for qualitaty log production and for mass assortments. The simplicity of these practice-oriented
models may foster the qualitative development of black locust management in Hungary and in some other countries where this tree species may gain greater acceptance by landowners and the forest industry. -
Dr. Ottó Orsós, the forgotten Hungarian pioneer in plant tissue culture
9-13.Views:170The knowledge of tissue culture deserves attention in respect of understanding the development of universal biology. This study intends to contribute to the past of the plant tissue culture by such data of the history of science which have been unprocessed so far. It seems that the life-work of the Hungarian biologist, Dr. Ottó Orsós is a missing and essential link between those early plant hormone researchers and the representatives of the pioneers of tissue culture schools who have contributed substantially to the development of the modern in vitro plant morphogenesis and plant cell biology. Orsós cultured kohlrabi tuber cubes on White culture medium in a sterile manner. This way, he could efficiently direct the in vitro morphogenesis of the kohlrabi, the regeneration of its shoot and root, and the formation and steps to subculture of pure callus tissues in 1938. He supported the correctness of its statements by means of detailed anatomical examinations. Orsós successfully rooted and aclimatized complete regenerated plants. We may as well call the above system — in remembrance of the creators of the original concept — "Haberlandt-Orsós model". Between the publishing of his main paper in 1938 and 2003, a period of 65 years has lapsed. On the occasion of this anniversary, we bow before this forgotten pioneer.
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Luminescence variations in cucumber (Cucumis sativus L.) leaves derived from different regeneration systems
50-52.Views:156Plants obtained from in vitro culture can show increased susceptibility to environmental stress conditions. In the process of their adaptation to natural conditions it requires monitoring of their physiological state. The methods used to check this phenomenon should estimate quickly and exactly the tolerance to suboptimal environmental factors. Such requirements are satisfied by the methods of measuring chlorophyll luminescence in vivo, e.g. fluorescence induction and delayed luminescence. The objects of our studies were cucumber plants regenerated from cultures of callus and embryogenic cell suspension, as well as the plants obtained from seeds. The plants derived from in vitro cultures displayed a poor physiological condition at the early phase of adaptation characterised by higher susceptibility both to stress caused by increased density of the light flux and low temperature (4 °C) in comparison with the plants obtained from seeds.
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Cultivation systems in plum orchards
125-129.Views:267In this review, we give an overview of recent cultivation systems in commercial plum orchards. The aspects of selecting the shape of the crown and the spacing of trees are discussed first and then growth characteristics and the structure of the crow are described in detail. Finally, the wound healing and regeneration characteritics of plum trees are detailed which have an influence on the applied pruning technique.
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Production of transgenic carnation with antisense ACS (1-aminocyclopropane44-carboxy late synthase) gene
104-107.Views:175Dianthus chinensis and Dianthus caryophyllus varieties were tested for shoot regeneration from leaf and petal explants and transformed with Agrobacterium tuniefaciens strains (EHA 105 and LBA 4404) harbouring an apple derived ACS cDNA in antisense orientation in order to reduce ethylene production and influence the ethylene dependant traits in carnation. After transformation regenerating shoots were selected on MS medium containing 50-75-100-125-150 mg/1 kanamycin and supplemented with 1 mg/1 BA, 0.2 mg/1 NAA. Transgene integration was proved by PCR analysis with npt II spcific primers followed by Southern hybridisation of DNA isolated from green shoots on medium containing 150 mg/1 kanamycin. Several putative transformants were subjected to RT-PCR in order to examine the npt 11 expression at mRNA level. Both the transformant and the non-transformant plants were potted into glasshouse to observe the effect of changed ethylene production on flowering time, petal senescence and vase life.
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Development of in vitro propagation system for Atriplex halimus L.
123-129.Views:217Explants 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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Comparative study of the vegetative and generative organs in pear varieties
21-25.Views:128An assortment of 17 pear varieties was examined in 2006 at Keszthely, Department of Horticulture, Georgicon Faculty of Agriculture, Veszprem University. The selected varieties were planted in 1980, grafted on seedling rootstock and represented the majority of existing pear plantations in Hungary. The main objective was the determination of suitability of the most important varieties for the purpose of intensive growing technologies even when grafted on vigorous seedling rootstock. The most important growing and fruiting characteristics of the varieties have been assessed and evaluated from the point of view of productivity. We stated that the relations of the trunk or the main axis to the lateral branches and fruiting structures are all subject to varietal effects and are valuable indices of the growing character. The quotient of the diameters of trunk and branch should be around 0.3-0.4, and the relative frequency of fruiting structures (Dárda, nyárs, vessző) meaning the ability of branching and regeneration associated with accurate pruning policies are decisive from the point of view of promising success.