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The tissue structure of the vegetative organs of strawberry (Fragaria moschata Duch®)
28-31.Views:232The 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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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:187The 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:251Immature 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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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:227Micrografting 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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Prevalence of banana diseases and post-harvest losses in Kenya, and biocontrol potential of arbuscular mycorrhizal fungi against Fusarium wilt
62-73.Views:251The demand for bananas (Musa spp.), which is ranked as the most important fruit crop in Kenya has been on the rise owing to both their dietary contribution and income generation. Meeting this demand has however been hampered by losses during production or post-harvest. This study assessed banana disease and post-harvest losses in leading producing counties in Kenya namely; Kisii, Nyamira and Embu. The study also assessed the efficacy of Rhizophagus irregularis in controlling Fusarium oxysporum f.sp. cubense. Structured questionnaires were used to collect data on post-harvest losses. Disease scoring tables, charts and photos were used to confirm observed symptoms and hence, disease occurrence and severity. AMF biocontrol efficacy experiment was conducted using tissue culture bananas grown in the greenhouse. The study revealed that most smallholder farmers were unaware of the causes or the prevalence of post-harvest losses. The findings also revealed a significant difference (p<0.05) in the severity of banana diseases across various cultivars from the three counties. The AMF treated bananas showed a significant difference (p<0.05) in plant height, total leaf area and chlorosis in comparison to other treatments. The study also revealed a reduction of Fusarium’s pathogenic effects including chlorosis, reduced leaf surface area and eventual necrosis.
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Studies on the alkaloid production of genetically transformed and non-transformed cultures of Lobelia inflata L.
65-71.Views:152The investigations of the growth and alkaloid production of cell suspension-, callus-, organized- and hairy root cultures from Lobelia inflata L. proved that these cultures are able to synthesize the characteristic piperidine alkaloids of the intact plant. Alkaloid precursor amino acids (Phe, Lys) and plant growth regulators affect not only the growth and differentiation of tissue cultures but also their secondary metabolism. The synthetic regulator Sz/I I combined with Phe increased the total alkaloid content considerably in callus- and organized cultures; regulator Sz/28 especially increased the lobeline content (in organized cultures in response to Lys, in callus tissues as a result of Phe application). With the aim of optimizing growth and alkaloid production of the genetically transformed hairy root cultures of Lobelia inflata L. we studied the effect of some growth regulators (NAA, IAA, kinetin) and precursor amino acids (Lys, Phe). The kinetin had inhibiting effect on the growth and lobeline production of the hairy roots. The IAA and NAA increased the biomass formation and lobeline production. The highest lobeline level was detected in tissues cultivated on hormone-free medium containing Phe.
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Ultrastructural and biochemical aspects of normal and hyperhydric eucalypt
61-69.Views:251Hyperhydricity was observed throughout in vitro multiplication phase of a Eucalyptus grandis clone. Ultrastructural approach of tissue and cell differentiation, izoenzyme patterns, binding protein (BiP) expression, and pigment content were performed. Hyperhydric tissues showed a reduction in cell wall deposition, reduction of membranous organelles, higher cell vacuolation, and more intercellular spaces than its normal counterpart. Additionally, several vesicles were present in hyperhydric cells suggesting the occurrence of organelle autophagy by autophagic vacuole. Lower pigment content, intercellular spaces on the epidermis and the induction of a molecular chaperone (BiP) were observed in hyperhydric phenotype. Evidences of schizolysigenous process of intercellular space formation are compatible with a stress condition. Although plastoglobulli were observed in normal and hyperhydric chloroplasts, they were more evident in the normal ones. Abnormal stomata also reflected a disruptive situation and morphogenesis disturbances which would difficult plant acclimatization. Further observation of the epidermis ultrastructure allows us to conclude that the presence of intercellular spaces on its surface may be constraining the recovery and development of hyperhydric plants. Similarly to BiP, other proteins such as esterase (EST), acid phosphatase (ACP), malate dehydrogenase (MDH) and peroxidase (PDX) are possible to be used as stress markers in in vitro conditions. Our results confirm earlier findings about negative effects of hyperhydricity on in vitro plant morphogenesis and ultrastructure, which in eucalypt is associated with a stressful condition contributing to lower propagation ratios.
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Effects of tuberization conditions on the microtuber yield and on the proportion of microtuber tissues
91-96.Views:124The production facilities of large-sized microtubers in three potato varieties (cv. Desiree, BorO, Gfilbaba) and the effects of the applied tuberization conditions on the proportion of microtuber tissues, especially on the perimedullary region were investigated in present work. In vitro tuberization was induced on explants with 2 or 5 nodes layered on MS medium supplemented with 8% sucrose. Induced cultures were exposed to short days (8 h) for 2 weeks, then to total darkness for further 11 weeks. For volume calculations of different tissue regions, the formula for ellipsoids (V=4/37c1/8/w2) was used. The number of large-size tubers (> 8 mm, up to 16 mm) reached 53%, 59% and 44% in cvs. Desiree, Giilbaba and Bore, respectively, which indicate that the size of microtubers could be increased by appropriate sucrose support and explant type. Microtubers produced on hormone-free medium have well-developed perimedullary region, and its volume rate seemed to be important in the final size of tubers. The increase in the rate of volume of the perimedulla was connected to the increase of tuber size until tubers reached 12 mm diameter. In microtubers larger than 12 mm in diameter, the volume rate of the pith was increased.
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Agrobacterium transformation of Rhodiola sp.: current status and limitations
37-42.Views:79The study of secondary metabolites has led to the discovery of new drugs for treating human diseases. However, consistent plant supply can be challenging, leading to the use of plant tissue culture techniques such as hairy root culture. Hairy roots have stable genetics, lateral branching, and can produce secondary metabolites, including alkaloids, flavonoids, and terpenoids. Research on hairy roots as a subject began in the late 19th century, and for the last four decades, hairy roots have been utilized for producing secondary metabolites and recombinant proteins. This article focuses on Rhodiola species - genus of perennial plants that belongs to the family Crassulaceae - and its potential as a source of secondary metabolites using hairy root culture techniques. Rhodiola sp. is widely distributed throughout the Arctic regions of the Northern Hemisphere, with several species having significant medicinal properties. The article discusses the possible use of hairy root cultures for the production of Rhodiola secondary metabolites, including salidroside and rosavins, which have demonstrated significant pharmacological activity in various studies. The use of elicitation and genetic engineering techniques to boost secondary metabolite production in Rhodiola hairy roots is also explored. Overall, the article highlights the potential of Rhodiola hairy root cultures as a valuable source of secondary metabolites with medicinal properties. However, despite some studies Rhodiola hairy root induction and culturing still remains highly unexplored.
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The role and current state of gene reservation of medicinal and aromatic plants in Hungary an overview
19-21.Views:195As it well known the decrease of biodiversity is a large problem all over the world. In case of medicinal plants, where the huge majority of drugs are collected from natural ecosystems, the sustainable, utilization of the populations and reservation of their gene-pools has an increased interest.
In Hungary, the major background of 'in situ' reservation of medicinal plant species, their natural plant associations and ecosystems is the official protection by law. Successful examples are known for the controlled utilization of medicinal and aromatic plant species grown in protected areas. Assuring the naturally occuring high degree of biological and chemical diversity of species is a special task in this field: only maintenance of valuable intraspecific races can form the real genetic basis of natural biologically active compounds.
Maintenance of chemotaxonomical gardens and gene bank collections (seed banks, tissue banks) as 'ex situ' methods of reservation is carried out on an extended range in Hungary. As the required information on storage and maintenance of many medicinal and aromatic plant species is yet missing, exhaustive research is carried out at both genebanks in Hungary, which are specialized for medicinal plant reservation (RIMAP- Budakalász, SZIU- Budapest).
Beside the static conservation methods, 'quasi-production systems' are intended to assure an up-to-date and economically possible way of dynamic reservation with sustainable utilization.
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Identification of ripening-related genes in strawberry fruit by cDNA-AFLP
33-41.Views:165An RNA fingerprinting study of strawberry receptacle and achene tissue was performed to identify candidate genes involved in fruit ripening. Quantitative cDNA-AFLP was used to detect differential gene expression in green, white, pink and red stages of fruit ripening. Based on hierarchical average linkage clustering the differentially expressed genes formed three major groups, genes expressed only in green receptacle, genes expressed mainly in white, pink and red receptacle, and in achene. 130 transcript-derived fragments (TDFs) were isolated and sequenced. Most TDFs did not show any homology to sequences with known functions, others were homologous to genes involved in oxidative stress response, signal transduction, regulation of development and cell-wall metabolism. Novel genes, so far not associated with strawberry ripening and ripening in general, were identified, such as genes encoding a bHLH protein, putative nitrilase-related protein, putative HD-zip protein. The differential pattern of gene expression draws the attention to the significance of ripening induced-or repressed promoters in strawberry fruit, whose isolation and characterization can be useful tool for functional genomics. For this purpose nine cDNA-AFLP fragments related either to ontogeny or senescing were completed with 5'UTR aiming at more precise annotation and future promoter isolation. Although tens of potentially important transcriptome changes were identified, the function of many ripening induced genes remain unknown.
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Susceptibility of sour cherry cultivars to isolates of Monilia laxa (Ehrenbergh) Saccardo et Voglino
83-87.Views:331In this study, the susceptibility of 7 commercially important sour cherry cultivars to Monilinia laxa was studied. Artificial inoculation was made with M. laxa isolates, which were isolated from different woody plants. Artificial inoculation was prepared in the laboratory and in the field. In laboratory, flowers of sour cherries while in the field, the two-year old twigs were inoculated in 2006 and 2007. According to results of stigmata inoculation, there were infection ability differences among the isolates originated from five different stone fruit host. Cultivars could be sorted into two susceptibility groups. In the field, twig inoculation in 2007 was made at blossom period and in 2007 at harvest. Seven sour cherry cultivars were inoculated with 8-day-old mycelial culture of M. laxa originated from sour cherry and almond. The agressivity and pathogenicity of the two isolates were measured by the degree of floem death: Results showed that year and phenological stage considerably influenced the degree of symptoms caused by the fungus. After artificial inoculation, tissue death progression was studied by fluorescent microscope. According to results, sour cherry cultivars were sorted into disease susceptibility groups. Susceptibility orders were identical to results on stigmata inoculation.
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Anatomical relations of root formation in strawberry
71-75.Views:211Anatomical relations of root formation are traced throughout the life cycle of the strawberry plant from the germinating seed up to the runners of the adult plant. Histological picture of the root changes a lot during the development of the plant. First the radicle of the germ grows to a main root, which makes branches into side roots and later adventitious roots are formed on the growing rootstock or rhizome. The anatomy of the different types of roots is also conspicuously different. First tiny branches appear relatively early after germination on the seedling's radicle, but soon the hypocotyl of the seedling thickens and develops side roots, which are already somewhat stronger. During this interval, the first true leaves are formed. The 4th or 5th of them being already tripartite, and the initiation of new roots extends into the epicotylar region of the shoot. The second years growth starts with the development of reproductive structures, inflorescences and runners starting from the axils of the new leaves. Near the tips of the runners below the small bunch of leaves, new root primordia are initiated. The tiny radicle of the germ develops a cortical region of 5-6 cell layers. Cells of the central cylinder are even smaller than the cortical parenchyma and include 3-4 xylem and 3-4 phloem elements as representatives of the conductive tissue. Roots originating from the shoot region are much more developed; their cortical zone contains 17-20 cell layers, whereas the central cylinder is about half as large. In the next year, new roots are formed at the base of the older leaves. These roots differ hardly from those of the last season in size and volume, however, they are recognised by colour and their position on the rhizome. The roots of the last year are dark, greyish-black, and grow on the lower third length of the rhizome, on the contrary, the new ones, on the upper region, are light brown. Roots starting from the shoot or rhizome are, independently from their age or sequence, mainly rather similar in size and diameter, thus being members of a homogenous root (homorhizous) system, i.e. without a main root. Plants developed and attained the reproductive phase develop in the axils of the leaves runners being plagiotropic, i.e. growing horizontally on the surface of the soil. The runners elongate intensely, become 150-200 mm, where some long internodes bear a bunch of small leaves and root primordia on short internodes and a growing tip. Runners do not stop growing, generally, further sections of 15-25 cm length are developed according to the same pattern, with small leaves on the tip. The growing tip of the runners is obliquely oriented, and small, conical root primordia are ready to start growing as soon as they touch the soil. The roots penetrate the soil, quickly, and pull, by contraction, the axis of the runner downwards, vertically, developing a new rhizome. The short internodes elongate a little and start developing adventitious roots. At the end of the growing season, the plantlets arisen on the rooted nods of runners are already similar to the original plants with homogenous root system. On the side of the adventitious roots, new branches (side-roots) are formed. The root-branches are thinner but their capillary zone is more developed being more active in uptake of water and nutrients. The usual thickening ensues later.
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Anatomical study of the leaves and petioles of scab resistant and susceptible apple cultivars
53-57.Views:178Anatomic 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:163Tissue 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.