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The tissue structure of the vegetative organs of strawberry (Fragaria moschata Duch®)
28-31.Views:233The tissue structure of the vegetative organs of strawberry (root, rhizome, stolon, leaf) is discussed in this paper. The authors stated that the root structure described by Muromcev (1969) and Naumann-Seip (1989) develops further from the primary structure. It grows secondarily and the transport tissue becomes continuous having ring shape. In the primary cortex of the rhizome periderm like tissue differentiates, but according to the examinations up to now, it does not take over the role of the exodermis. The exodermis is phloboran filled primary cortex tissue with 3-4 cell rows under the rhizodermis. The development of the transport tissue of the petiole is also a new recognition. In the lower third of the petiole the transport tissue consists of 3 collaterally compound vascular bundles. In the middle third there are 5 bundles because of the separation of the central bundle and in the upper third of the petiole 7 bundles can be observed because of the ramification of the outside bundles. Therefore attention must be taken also in the case of other plants at making sections. There might be confusions in the results of the examinations if the number of bundles increases in the petiole. The tissue structure might vary depending on the origin of the tissue segment.
The palisade parenchyma of the leaf blade has two layers and it is wider than the spongy parenchyma. Among the 5-6-angular cells of the upper epidermis do not develop stomata while in the lower epidermis there are a fairly lot of them.
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Dr. Ottó Orsós, the forgotten Hungarian pioneer in plant tissue culture
9-13.Views:195The 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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In vitro plant regeneration from immature embryo axis and cotyledons of common bean (Phaseolus vulgaris L.)
93-97.Views:139Phaseolus 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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Evaluating vase life and tissue structure of some compositae (Asteraceae) species
87-89.Views:197The vase life of cut flowers and effects of various chemicals was examined with the help of a pulse treatment. According to the results using of chemicals (preservatives, disinfectants as well as blocking of synthesis of ethylene) is ineffective if it is used after seeding This shows the great importance of harvesting time.
Using 8-HQS or l-MCP + 8-HQS proved to be the best for vase life in most of the samples. Using these materials did not prevent the appearance of air bubbles in the stems and absorption could be observed continuously.
To examine the tissue structure reaction of chemicals stems were stained with toluidin-blue, and high of absorption was measured. It was found that in cases, when absorption was bad, small air bubbles blocked the xylem vessels.
All the species examined (Aster linosyris, Achillea collina, Aster novi-belgii, Inula britannica, Solidago canadensis, Inula ensifolia, Senecio jacobea) show similar reactions to chemicals because they are the members of the same family.
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General defense system in the plant kingdom III.
45-54.Views:184Our observations regarding the symptoms not fitting into, significantly differing from the hypersensitive defense system, which we noticed during the judgment of several plant species, symptoms provoked on several million plants have constituted a unified entity. They have provided evidence for the existence of a different plant defense system. We called this so far unknown basic response of plants to biotic effects as general defense system. This system defends them from the attack of numerous microbe species in the environment.
The evolutionary intermediate phase between the general and the specific, the two defense systems is the susceptible host—pathogen relation. The vertical resistance system of plants escaping from the susceptible host—pathogen relation, based on specific hypersensitive reaction also suggested the existence of a more original, general defense system and the susceptible host—pathogen relation developed as a result of the collapse of that system.
The evolutionary relation of the two defense systems is proved by the only recessive inheritance of the older general defense system and in the majority of cases dominant hereditary course of the specific defense system. In our experiences, the modifying genes of the recessive general defense system, in most cases, are behind the specific defense systems, which are known to have monogenic dominant hereditary course and react with hypersensitive tissue destruction. This seemingly striking genetic fact is explained by the following: the general defense system less dependent on environmental effects regulates much faster pathophysiological reaction than the specific resistance genes strongly dependant on environmental effects coding dominant hypersensitive reaction.
The general and specific defense reactions, the processes excluding the microbes attacking plants with compacting of cell growth and tissue destruction, which mean two opposite strategies, building on and regulating each other constitute the entity of resistance to plant disease.
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Model experiments for establishment of in vitro culture by micrografting in apple
47-49.Views:228Micrografting 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.
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General defense system in the plant kingdom
79-84.Views:147The goal of plant breeders is to improve the resistance of crops against virus, bacterium and fungus pathogens was easiest to achieve by selection for phenotypes displaying the hypersensitive reaction. The resistant plant of that type keeps its health by preventing or delaying the systemization of the pathogen by destruction of cells and tissues of variable size or amputation of the contaminated organs. The faster the reaction of the host plant is the more efficient and economical is the defense, since the extent of tissue destruction decreases proportionally with the speed of reaction.
During a breeding program for resistance carried out on several plant species, mainly vegetables over thirty years, also an alternative defense reaction has been experienced, which fundamentally differs from the hypersensitive reaction. In that reaction the cells and tissues of the host plant being exposed to the pathogen do not die, on the contrary they hinder systemization of the pathogen by tissue thickening. An additional significant difference is that on the contrary to hypersensitive reaction this reaction is less host- or pathogen-specific and works excellently even at high temperature (over 40 °C).
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Vegetative and micropropagation potential of Piper guineense (Schumach and Thonn)
29-36Views:189The 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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The in vitro and in vivo anatomical structure of leaves of Prunus x Davidopersica ‘Piroska' and Sorbus rotundifolia L. ‘Bükk szépe'
92-95.Views:252Immature in vitro leaves showed similar structure of the mesophyll tissue to the immature field-grown (in vivo) leaves of Prunus x davidopersica `Piroska'. Mature leaf anatomical characteristics of in vitro plantlets differ from the field-grown plants. The mesophyll tissue of in vitro plantlets were thinner than the in vivo plants and consisted of only one layer palisade parenchyma, the shape of the cells and the structure of spongy parenchyma basically differed from the field-grown plants. In the case of Sorbus rotundifolia similar anatomical differences were found both in vitro and in vivo as in the case of Prunus x davidopersica `Piroska'.
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Anatomical study of the leaves and petioles of scab resistant and susceptible apple cultivars
53-57.Views:179Anatomic studies have been performed on the leaf blade, petiole and annual shoot on six apple cultivars by means of scanning electron as well as light microscope. Four of the cultivars examined are resistant to scab (Florina, Freedom, MR-10, MR-11), whereas two of them are susceptible (Jonathan and Idared). Preliminary results suggest that differences in the width of cross sections of leaf blades, in hairyness, in the shape and size of epidermal cells, moreover, in the cross sections of petioles and shoots are considerable. Some of the anatomical properties seem to be correlated with scab resistance or susceptility of the respective cultivars. Therefore, further studies extending to other cultivars may corroborate our claims to find causal relations between anatomical traits of the leaves and disease, especially scab resistance of apple cultivars.
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Callus induction on standard type Cymbidium cultivars
108-110.Views:164Tissue cultured Cymbidium PLBs (protocormlike body) were used as starting material to induce embryogenic callus which could serve as objects of genetic transformation. We obtained callus using two methods. The first method was culturing the PLB segments for one month in liquid MS medium in the presence of 0.5 mg/1 benzyladenine and 0.05 mg/1 naphtylacetic acid followed by cultivation on the same composition solid medium with 0.5 g/l activated charcoal for an additional month. Callus formation was observed on 30% of the explants. The second way was to propagate the PLB segments on solid MS medium supplemented with 1 mg/1 thidiazuron. In these cultures we also observed callus formation on 20% of the explants.
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Frost induced changes in enzyme activities and carbohydrate content in the spurs of some pear cultivars during the dormancy
41-44.Views:226Frost tolerance of pear cultivars was checked after artificial cold treatment in 2003-2005. Limbs collected during the endodormancy were exposed in a climatic chamber for 24 hours to —25; —28 °C, while those collected in the ecodormancy were kept at —15 and —18 °C. Frost damages of buds were registered according to a visually defined scale, then peroxidase (POD), polyphenol oxidase (PPO) enzyme activities and carbohydrate contents were checked in buds and spur-part below the buds. POD activity of untreated control in tissue below buds was higher than in the buds, which were increasing continuously during the endodormancy and decreased at the end of the ecodormancy. During endodormancy, cold treatment of —25 and —28 °C effected different changes of enzyme activity in buds of the cultivars. In the ecodormancy, enzyme activities increased after a cold treatment of —15 °C, whereas the activities decreased significantly after —18 °C. `Kaiser' — susceptible to frost — with its higher values of both enzyme activities marked out from other cultivars, which is correlated with its stress response. Changes in carbohydrate components — especially in glucose — of buds monitored well the different stress responses of tolerant and resistant pear cultivars induced by frost stress.
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Study of different factors of grapevine regeneration systems and genetic transformation
33-36.Views:220The 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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Shoot induction and plant regeneration from cotyledon segments of the muskmelon variety "hógolyó"
61-64.Views:151Cotyledonary 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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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:193Vine 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:226In 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.