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Micropropagation of Rudbeckia hirta L. from seedling explants
105-108Views:195We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L seedlings. We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps. First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times. For the propagation of seedling meristems, the combination of half-strenght solid Murashige and Skoog (1962) culture medium containing 10 mg/1 of kinetin or 2 mg/I of kinetin + 0.1 mg/1 of 2iP proved to be the most suitable. The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/1 of IAA. After four weeks of incubation we obtained elongated shoots which we separated and inoculated into a new culture medium and we obtained elongated roots. The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation. By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respect of breeding.
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High-velocity microprojectile mediated DNA delivery into Phaseolus vulgaris callus cells
99-102.Views:129We report the method for the establishment of rapidly growing callus cultures of Phaseolus vulgaris and the conditions required for efficient transformation using high velocity microprojectiles and high level of transient gene expression. Using hypocotyl explant and vertical culture on B5 medium with lmg/1 kinetin and 2 mg/1 2,4-D, we can recommend to get a rapidly growing callus from bean which is a good starting material to introduce foreign DNA into bean cells. The GeneBooster particle delivery system was used for the bombardment of bean callus and the Hgm resistance gene (Hgmr) was used as a selectable marker gene. 25mg/I hygromycin (Hgm) concentration was sufficient to kill the control callus. We used the standard physical factors, the appropriate pressure of N2 gas for the bombardment of the callus tissue, the shooting distance and the size of tungsten particles used as microprojectiles. Selective and nonselective tests were made by transferring the healthy green and white calluses, subcultured for 4 months on selective and nonselective medium. Several Hgm resistant calli had been obtained. Selective pressure was maintained over a period of 10 months.
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The evaluation of grape vine decline pathogens in the experimental field of the Georgikon Faculty of Agriculture in Cserszegtomaj
19-22.Views:192Vine decline causes important economic loss in viticulture, especially in longer view. Several causal pathogen were described lately, however little is known about the etiology or epidemiology of these pathogens on grapevine rootstock. It is well known that these diseases affect grafted and rooted grapevines and are not specific to any scion-rootstock combinations. Our aim was to determine what pathogens are presents in the experimental field, especially causal agents of the rootstock decline. Different grapevine rootstocks and scion varieties were tested in our trial. Isolations were made from the wood tissue and pathogenity tests were done with isolated Cylindrocarpon destructans. The possibility of infection during the propagation process was studied as well. Most commonly Cylindrocarpon sp. and Phomopsis sp. species were identified from the examined varieties. Cylindrocarpon destructans was able to spread to apical (shoot) and basal (root) direction from the point of infection with uneven speed. Callus development is not inhibited by the fungi causing the leaf symptom of the vine decline. Shoot development is reduced if unhealthy parts are grafted to each other.
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Improved clonal approaches to growing black locust (Robinia pseudoacacia L.) in Hungary: a case study
53-56.Views:286In Hungary black locust (Robinia pseudoacacia L.) is considered as an important exotic stand-forming tree species and due to climate change effects its importance is increasing in many other countries. It has some desirable characteristics from both the practical and research standpoints. As a result of a partly new black locust selection programme new black locust clones were improved and a technology was developed for mass clonal micropropagation of juvenile trees. Clone trials with micropropagated plants were established in the country for evaluating the juvenile growth and the stem form of promising black locust clones under marginal site conditions. Significant differences (P<5%) were found for stem form value which partly verified the genetic gain of the selected clones against the common black locust. It was also proved that tissue culture could offer partly new prospects for the rapid mass cloning of selected genotypes.
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Attempting Regeneration from Cultured Cotyledons and Plant Regeneration from Cotyledonary Nodes in Common Bean (Phaseolus vulgaris L.)
57-60.Views:167Dry 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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Anatomical relations of the leaves in strawberry
81-84.Views:225In the present study histology of the leaves of strawberry (Fragaria ananassa Duch.) variety Elsanta was the objective, which has been performed with the beginning of seedling stage, cotyledons, primary leaves and later true leaves, first cataphyll of the runner shoot as well as the bracteoles of the inflorescence. Structures of the leaf blade, the upper and lower epidermis, the petiole have been also observed. The leaf blade of cotyledons already contains a typical palisade as well as spongy parenchyma tissues, i.e. being bifacial showing a structure similar to that of the true leaf. However, the petiole displays differences from the true leaf. There are a narrow (4-5 layer) primary cortex and a tiny central cylinder. Primary leaves bear already hairs on the adaxial surface and the transporting tissue-bundles are recognised in cross sections having a "V" shape. The first true leaf composed by three leaflets is of a simple structure showing characters reminding of cotyledons and primary leaves. Leaves of intermediate size continue to grow, whereas their inner anatomy changes dramatically. In the central region of the leaflets, near to the main vein, a second palisade parenchyma appears, further on, transporting tissue bundles are branching in the petiole. Collenchyma tissues enhance the stiffness and elasticity of the petiole. Older true leaves develop thick collenchyma tissues around the transporting bundles being represented by increasing numbers. The doubled palisade parenchyma layers of the leaf blades are generally observed. The cataphylls of the runners have a more simple structure, their mesophyll is homogenous, no palisade parenchyma appears. It is evident that leaves grown at successive developmental stages are different not only in their morphological but also anatomical structure. There is a gradual change according to the developmental stage of the leaves.
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Factors affecting rhizogenesis in vitro and acclimatisation of three cherry rootstocks
40-46.Views:135Factors 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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Clonal selection of black locust (Robinia pseudoacacia L.) in Hungary: a review
153-56.Views:196Black locust (Robinia pseudoacacia L.) is the most important fast growing stand-forming tree species in Hungary. Its importance is increasing in many other countries, too. As a result of a new selection programme 13 black locust clones have been improved for setting up clones trials and seed orchard. In 2003 five of them (R.p. `Bácska', `Homoki', 'Szálas', `Oszlopos' and `Vacsi') were registered as cultivarcandidates. Tissue culture method has proved as a suitable mean of propagating superior individuals. The micropropagated plants have been growing successfully in the clone trials.
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General defense system in the plant kingdom II.
69-71.Views:136In addition to successes achieved in certain varieties in resistance breeding based on a defense reaction of host plants involving hypersensitive tissue destruction, resistant varieties putting a very strong selection pressure on pathogens have selected more and more aggressive types of pathogens. The never-ending race between plant and pathogen resulting from this can only be controlled by a defense system characterised by a different strategy. In each of the plant species that we bred a defence system was found, which contrary to hypersensitive reaction strives to keep the tissues at all costs and is not pathogen specific. This is implied in the term general defense system.
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Jerusalem artichoke (Helianthus tuberosus L.): A review of in vivo and in vitro propagation
131-136.Views:590Jerusalem artichoke (Helianthus tuberosus L.) is an old tuber crop with a recently renewed interest in multipurpose improvement. It is a perennial tuberous plant rich in inulin and is a potential energy crop. During food shortages in times of war Jerusalem artichoke received more attention by scientists and farmers because of its multiple uses as a vegetable, medicinal plant, forage plant and source for biofuel. The energy crisis of the 1970s motivated research on Jerusalem artichoke for biofuel as the aboveground plant biomass and the tubers can be used for this purpose. There are different methods to propagate Jerusalem artichoke using tubers, rhizomes, slips (transplants derived from sprouted tubers), stem cuttings, seeds and tissue culture. So, this review was presented to highlight on propagation of Jerusalem artichoke via in vivo and in vitro techniques.
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Anatomical study of the bud union in „Chip" and „T" budded 'Jonagold' apple trees on MM 106 rootstock
27-29.Views:412The traditional methods for vegetative propagation of apple and its varieties are the T-budding, and the winter grafting, but this latter way is a difficult and expensive procedure.
In our experiment carried out in the Fruit Tree Nursery Soroksár, the healing process of chip- and T-budded apple trees 'Jonagold' on MM 106 rootstock was studied.
The budding (T- and Chip-) was made in the first week of August, samples for microscope examination were taken monthly after this time until leaf fall.
The investigated part of plants was made soft with 48 % HF (hydrogenfluoride), then cross and longitudinal section were made and examined by microscope.
Based on analysis of microscope pictures in case of Chip-budding, it was established, that development had started quickly after budding on the rootstock and scion too. But the callus originated almost entirely from the rootstock tissue as new parenchyma cells fills the gap between the two components of graft (scion and stock), becoming interlocked and allowing for some passage of water and nutrients between the stock and the scion. This quantity of callus in case of T budding was under the scion buds larger, than the Chip-budded unions, where the thickness of callus mass is uniformly thick round the chip. The large mass of callus pushes the scion bud outwards from the shoot axis, which later results in a larger shoot-curvature above the bud union.
Following this process on the Chip-budding it can be observed also, that a continuity of the cambium is established between bud and rootstock. Then the newly formed cambium started typical cambial activity, forming new xylem and phloem.
Later the callus begins to lignify, and it is completed within about 3 months after budding.
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Bean tissue culture and genetic transformation with Agrobacterium
32-35.Views:153In this paper we report the establishment methods of a rapidly growing callus culture of Phaseolus vulgaris bean as well as the conditions required for a high level of transient gene expression using Agrobacterium-mediated transformation. A vector is containing both the lindan-resistance gene as a selectable marker, and GUS gene as a screenable marker. By using hypocotyl explant and vertical culture on B5 medium supplemented with 1 mg/1 kinetin- and 2,4-D 2 mg/1 and subcultured every 3-4 weeks, we can recommend to get a good and much callus from bean. This will help in introducing foreign DNA into callus cells. One strain of Agrobacterium carrying plasmid as vector for introducing foreign DNA into plant cells was used. At different concentrations of lindan; 3, 4 and 4.5 mg/I, the transformed Maxidor callus survived and grew over a period of 6 month and subcultured every 3-4 weeks, but the control callus died. Callus were assayed for GUS activity to confirm the expression of the GUS gene using the histochemical assay test. The GUS gene was also correctly expressed in callus cultures grown on 4mg/I lindan-selected medium, the typical blue colour in the histochemical assay using the X-gluc as substrate. But the control, non-transformed callus was not able to grow in the presence of lindan, neither showed a positive reaction in the in vitro assays.
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Biotechnologies for ornamental plants: some insights to the Brazilian productive chain
51-59.Views:282The world industry of ornamental plants is under wide transformations regarding the changes in the cultivation, management, postharvest technologies and the use of new (bio)technologies to assist the improvement and development of new varieties. This industry worth more than US$ 20 billion, but Brazil shares only 1% of this value. To compete at this market with permanent technological innovation, the country needs to be tuned in with the progresses in the biotechnologies applied to the productive chain of ornamental plants. In this revision, we analyze the Brazilian and world ornamental market emphasizing the role of the biotechnologies in the modernization and increase of the competitiveness of the sector.