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Rhizogenesis in in vitro shoot cultures of passion fruit (Passiflora edulis f. flavicarpa Deg.) is affected by ethylene precursor and by inhibitors
47-54.Views:169The effects of the ethylene precursor ACC and two inhibitors, AgNO3 and AVG, on root formation were tested in in vitro shoots of passion fruit (Passiflora Midis f.flavicalpa Deg.). The organogenic response was assessed on the basis of percentage of shoot-forming. roots, root number and length. The time course of ethylene production was also monitored. ACC inhibited root formation by delaying root emergence and increasine, callus formation at the basis of the shoots. In addition, ACC caused a marked increase in ethylene production, coupled to leaf chlorosis and senescence with lower rooting frequencies, number and length of roots. IAA supplementation increased ethylene production. Both ethylene inhibitors, AgNO3 and AVG, at appropriate concentrations reduced callus formation at the basis of shoots. AVG increased the number of roots per shoot, but drastically reduced length of differentiated roots. Regarding to leaf pigments, ACC promoted a marked reduction on carotenoids and total chlorophyll, whereas AVG and AgNO3 delayed explant senescence and pigments degradation, not differing from IAA supplemented and non-supplemented control treatments. The results confirm previous reports on the beneficial effects of ethylene inhibitors on in vitro rooting and suggest its reliability to be used as an alternative approach to evaluate sensitivity of Passiflora species to ethylene.
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Anatomical relations of root formation in strawberry
71-75.Views:173Anatomical 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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Growth Rings in the Stem of Thuja orientalis L.
22-27.Views:140During the winter dormancy period of 1998-99, the differentiation process of xylem and the formation of annual and pseudo-annual growth rings was studied in sections of the central stem of Thuja orientalis of different age (1-14 year old), starting from the top (the tip of leader shoot) towards the bottom. In the apical 1-2 cm portion of the one year-old leader shoot, only the protoxylem was formed by the end of vegetation. The protoxylem elements appeared first in 6 bundles than gradually merged into 2 semiquadrangular (triangular) bands (each containing 3 protoxylem bundels) around the pith. In this stem portion, the pith was cross-formed first and became gradually flattened at the lower stem parts, following the generally flattened shape of the stem and the respective facial and marginal position of leaves. A continuous xylem ring (with formation of metaxylem elements) apeared 3-5 cm below the shoot tip. In fact, it was the stem part where a "real" annual ring was formed by the end of vegetation. The first pseudo-annual rings were observed 16 cm below the top. The "regular" annual rings were completely continuous all around the stem, consisted of strongly flattened in radial direction thick-walled latewood tracheids and had a distinct border (demarcation line) at the end of the year. The "pseudo-annual" rings formed incomplete dark rings or semicircular bands within the earlywood. They were composed of tracheids with thick cell walls but somewhat wider radial diameter than those of the "real" annual rings, and the border between their outside margin and the next earlywood was less distinct.
In the xylem of two year old stem portion, the innermost central annual ring appeared not at the transition zone between the current and the former years of growth, but about 2 cm lower. Above that, only the thick-walled bundles of the former years protoxylem were found. Down the stem, the older sections showed similar features: the next annual ring appeared always somewhat lower than the borderzone of the given and the former years growth. The "pseudoannual" rings (or more correctly the growth rings) continued regularly to appear in the lower (older) sections of the stem as well. They were found untill the age of up to 14 years (the bottom of the studied plants). Their number was 3-4 per year first, than (from the 5th annual ring counted from the centre) decreased to 3,2,1, and in the youngest outer part of the xylem there was no pseudo-annual ring at all. The development of pseudo-annual rings was usually more marked on the thicker (more branched) than on the thinner side of stem. Stems older than 14 years were not studied.
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Relaco: a product to reduce water shoot formation around pruning wounds in pome fruit trees
30-31.Views:108Larger cuts made during the winter pruning of pome fruit orchards a number of serious problems: the wounds serve as entrances for several branch-infecting pathogens, the numerous water shoots formed around the cut edges consume assimilates, shade the developing fruits, make the canopy dense, reduce the effectiveness of the pest management and cause extra work during next seasons pruning work. These problems can be eliminated by using proper wound protectants, like Relaco, a new Romanian wound protectant to cover larger pruning cuts of apple, pear and quince trees.
The treaments by Relaco resulted in a significant decrease in both the number and total length of water shoots formed around the cut egdes.
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The effect of different biostimulators on morphological and biochemical parameters of micropropagated Hosta ’Gold Drop’
22-29.Views:246During in vitro multiplication of Hosta ‘Gold Drop’, 20 g l-1 sucrose, 5.5 g l-1 agar and 4 concentrations (0.1-0.8 ml l-1) of Ferbanat L, Kelpak, Pentakeep-V were added to half-strength Murashige and Skoog (MS) basal medium. As compared to the control and other biostimulators, plants with lower peroxidase activity, larger fresh weight, more, longer shoots and roots, larger leaves were developed on medium containing Kelpak. The best concentration was 0.4 ml l-1 for in vitro rooting, shoot formation, plant weight and ex vitro chlorophyll, carotenoid level, peroxidase activity. Pentakeep was the less efficient biostimulator, increasing of its concentration mostly decreased root and shoot values (furthermore, abnormal callus formation was observed, as non-wanted effect), chlorophyll content and sizes (length, width) of leaves, not only during in vitro propagation but also (as after-effect) acclimatization because of the high mortality and weakly developed survivor plants.
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Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)
15-12.Views:274In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants.
Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of 0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance . -
Shoot induction and plant regeneration from cotyledon segments of the muskmelon variety "hógolyó"
61-64.Views:143Cotyledonary segments of the casaba type muskmelon variety "Hógolyó" were used to induce organogenesis. Fifty different hormone combinations were applied to enhance the induction of shoot formation on the edge of the segments. The phases of organogenesis were followed with light- and scanning electron microscope. Shoot induction was achieved with high frequency. The shoots were transferred to hormone free media for root induction. The rooted plantlets were planted out to soil.
NAA was feasible and the method can be applied in transformation experiments.
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Security of growing habits and bud formation of German sour cherry varieties
45-48.Views:203Intensive sour cherry production is concerned to find the most productive varieties under special growing conditions. High planting density, adapted to manual picking or on the other hand to mechanical harvesting. Almost as important is the prolongation of the ripening season by enlarging the choice of varieties. Unfortunately, three of our four leading commercial varieties are ripe almost at the same date.
Therefore, new varieties ought to be examined thoroughly. In the Pallag Experimental Station of the Debrecen University, five European varieties have been grown (’Schattenmorelle’, ’PI-SA 12,100’, ’Jade’, ’Gerema’, ’Achat’) and a Hungarian one, ’Debreceni bôtermô’ used as a check for the experiments to compare their growing and yielding habits in 2010. The plantation was three years old, standing on Prunus mahaleb rootstocks, in high density (5 x 2 m) and trained to slender spindle crowns. The results are proving that some of the varieties in
question are suitable to prolong the harvesting season. Growing habits and yields of the varieties related to the variety ’Debreceni bôtermô’ were similar or even better as ’Jade’, ’Gerema’ and ’Achat’. The ’Jade’ excelled with its vitality and ’Gerema’ with its generative character. Further studies are expected to prove the utility of
one of them at least to enlarge the ripening season on the fruit market. -
Investigation of the in vitro regeneration of mericlones in the caribe variety of carnation
87-89.Views:131In vitro culture conditions were experimented for the relatively sensitive, but very esthaetic "Caribe" variety of carnation with uniformly dark violet flowers. Regeneration of new plants from shoot apex meristems can be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. Callus formation as a prerequisite for the induction of somaclonal variability can be achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Acclimation of the obtained mericlones to the ex vitro conditions was also evaluated.
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Characterization of sunburn damage to apple fruits and leaves
15-20.Views:453The specific conditions of the formation of three different types of sunburn (sunburn browning, sunburn necrosis, and photooxidative sunburn) have been recently characterized on apple fruit. However, no information is still available on leaf damage. Therefore, the aims of this study were i) to extend the knowledge on fruit damage, ii) characterize leaf damage and iii) find relationship between fruit and leaf damage. The observations were made on 586 apple accessions in a gene bank orchard located in Hungary. The incidence of the three different types of fruit symptoms were recorded and based on the visual symptoms, two different types of leaf sunburn (sunburn yellowing and sunburn necrosis) were characterized. The most frequent type of fruit sunburn observed was sunburn browning. Photooxidative sunburn was found for less number of accessions, and only some accessions were affected by sunburn necrosis. Fruit were far more susceptible than leaves; (>60%) of the examined accessions were affected by fruit damage and (<3%) by leaf damage. Although a large number of accessions were affected, the percentage of fruit damaged within accessions was not that excessive; ~6% of the fruit assessed showed the symptoms of sunburn browning. Significantly fewer fruit were damaged by sunburn necrosis (~1%) or photooxidative sunburn (~1.4%) than sunburn browning. The percentage of leaves damaged within accessions were simlarly very low (~1%). Close relationship between fruit and leaf damage was found. Accesions with relatively heavily sunburned leaves usually had severe fruit damage as well. Leaves showing sunburn symptoms were usually closely located around those fruit which were sunburned severely. Leaf damage of sunburn was found on spur leaves in a great majority of the accessions damaged, shoot leaves did not seem to be susceptible to sunburn.
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Development of in vitro propagation system for Atriplex halimus L.
123-129.Views:217Explants excised from adult shrubs were surface sterilized and cultured on Murashige and Skoog (MS) basal medium in the presence of plant growth regulators (PGRs) at different concentrations. A high multiplication rate of 7.2-fold was achieved every four weeks on MS medium supplemented with 4.44 μM BA, 0.49 μM IBA and 0.58 μM GA3. Rooting was achieved with 73% efficiency within 2-4 weeks on agar-gelled MS basal medium free of PGRs. Rooted plantlets were gradually acclimatized to field conditions over 5-6 weeks with 65% efficiency. For in vitro selection for salt tolerance, MS medium was supplemented with increasing concentrations of NaCl ranging between 25 and 1000 mM. This study has demonstrated that in vitro shoots could tolerate up to 600 mM NaCl with optimal growth at 200 mM, while higher concentrations of NaCl affected growth negatively. Growth and shoot number decreased with increasing NaCl concentration with all plantlets died at 1000 mM NaCl.
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Influence of different growth regulators on the in vitro morphogenesis of an ornamental variety of carnation
55-57.Views:141Callus formation, as a prerequisite for the induction of somaclonal variability, was achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Regeneration of new plants from shoot apex meristems could be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. These in vitro treatments may contribute to a more efficient micropropagation of the Rimini variety of carnation.
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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:377The 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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In vitro regeneration from cotyledons of watermelon
96-98.Views:166Cotyledonary segments of five different genotypes of watermelon were used to induce organogenesis. Five different hormone combinations were applied to enhance the induction of shoot formation on the surface of the segments. The phases of organogenesis were followed with light and scanning electron microscope. Shoots were obtained after four weeks, then the shoots were transferred to hormone free medium for root induction.
This method of regeneration can be applied in transformation experiments. GUS histochemical assay was made to check the expected success of using Agrobacterium for the transformation.
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Dr. Ottó Orsós, the forgotten Hungarian pioneer in plant tissue culture
9-13.Views:170The knowledge of tissue culture deserves attention in respect of understanding the development of universal biology. This study intends to contribute to the past of the plant tissue culture by such data of the history of science which have been unprocessed so far. It seems that the life-work of the Hungarian biologist, Dr. Ottó Orsós is a missing and essential link between those early plant hormone researchers and the representatives of the pioneers of tissue culture schools who have contributed substantially to the development of the modern in vitro plant morphogenesis and plant cell biology. Orsós cultured kohlrabi tuber cubes on White culture medium in a sterile manner. This way, he could efficiently direct the in vitro morphogenesis of the kohlrabi, the regeneration of its shoot and root, and the formation and steps to subculture of pure callus tissues in 1938. He supported the correctness of its statements by means of detailed anatomical examinations. Orsós successfully rooted and aclimatized complete regenerated plants. We may as well call the above system — in remembrance of the creators of the original concept — "Haberlandt-Orsós model". Between the publishing of his main paper in 1938 and 2003, a period of 65 years has lapsed. On the occasion of this anniversary, we bow before this forgotten pioneer.