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Jerusalem artichoke (Helianthus tuberosus L.): A review of in vivo and in vitro propagation
131-136.Views:903Jerusalem 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:693The 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:330In 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:433The 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.
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Identification of the apple firmness: two case studies
32-35.Views:229Firmness tests were performed with peeled and entire fruits of Elstar and Jonagold apple cultivars for identification and comparison. The normal distribution of the tested population was acceptable (level: 95%). The green and the red sides did not show differences within the cultivar but they were different in firmness. There was not significant difference between the flesh tissue firmness values, however the firmness of the entire (not peeled) fruits was different. This result was caused by the effect of the peel. The variability of the firmness with Jonagold was caused by the peel, but such a result was not found with Elstar. The test of the peel effect would be interesting with different cultivars and a sequence according to the firmness can be estimated.
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The importance of pathogen-free grapevine propagation material in regards to clonal selection and rootstock breeding in Germany
11-13.Views:347High performing propagation material is essential for a reliable and economical production of quality grapes. Apart from genetic aspects pathogen-freedom is of prime importance in propagation material. In particular virus diseases cause major yield and quality losses and reduced longevity. This is also reflected in the current EU legislation, which focuses on the most common and dangerous viruses: GFLV, ArMV, GLRaV-I and GLRaV-III.Apart from these, locally occurring pathogens, e.g. phytoplasms or agrobacterium, are important as well and should not be present in propagation material. There are several ways to develop pathogen-free clones. Starting with already pathogen-free material is certainly the easiest case, but might not be feasible in local varieties with small acreages and limited vine numbers. In these cases the elimination of pathogens is required, either by heat therapy, tissue culture or somatic embryo genesis.
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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:775The 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:270The 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:495Hyperhydricity 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:232The 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:329The 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:380As 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:298An 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:494In 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:508Anatomical 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:340Anatomic 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:346Tissue 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.
