Mass propagation of 5 newly introduced Hosta varieties was carried out by the method of micropropagation. Our aim was to determine exact variety specificity after the micropropagation period in the pattern of peroxidase isoenzymes by isoelectric focusing in pH 3-9 range and to determine that phenological phase of mother plant in which...the isoenzyme pattern of mother plant can safely be comparable to the isoenzyme pattern of micropropagated descendants. The isoenzyme patterns of descendants were similar to the mother plants of the same hybrid lines. The older leaves seemed to be not so suitable for examination than newly developed ones despite of the higher activity of peroxidase enzymes. There were big differences in isoenzyme patterns of leaves in different phenological phases. With this quick and easy method Hosta varieties could be selected already in the very early stage of micropropagation.
We 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 solut...ion 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.
This paper gives an outline of micropropagation of Pontederia lanceolata. Pontederia l. is a widely used aquatic plant, therefore there is an increasing demand for them, which can be satisfied only by in vitro culture. Research was carried out to find the best nutrient media conditions for micropropagation and acclimatisation of Po...ntederia lanceolata.
In 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 progr...amme 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.
The effect of seven concentrations of two carbohydrate sources were compared to determine the best source and the most suitable source and concentration for micropropagation of some Hosta cultivars: H. 'Gold Haze', H. 'Gold Drop' and H. 'Dew Drop'. 0, 5, 10, 20, 30, 40 and 50 g/1 sucrose or glucose were added to a MS...basic medium supplemented with 3 mg/1 kinetin and 0.1 mg/1 IAA. For 'Gold Haze' 40 g/1 sucrose proved to be the best source and concentration, the proliferation ratio was 15 shoots per explant. Thirty g/1 sucrose concentration was the optimum for 'Gold Drop', the proliferation rate was 14.6 shoots per explant. In 'Dew Drop,' the best results were obtained with 30 g/1 sucrose but 40 g/l sucrose gave good results too. Both cultivars rooted well on these media. On glucose containing media, very low propagation rates were found in all concentrations and all examined cultivars.
Black 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.
Tissue 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 0.0...5 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.
A simple and reliable procedure for in vitro propagation of an orchid Dendrobium strongylanthum Rchb.f. was studied. Protochorm was induced from seed explants on 1/2 MS medium supplemented with 0.2 mg/L NAA. A mass of protochorm could be multiplied on proliferated medium of 1/2 Ms containing 0.5 mg/L V-6-BA. And bud differenti...ation of green global body was cultured in the same media, 2-2.5 cm shoots were formed after 30 day of culture. Addition of mashed banana and 0.5 mg/L NAA to 1/2 MS medium promoted root formation and vigorous growth. The plantlets were acclimatized and transplanted to compound materials of humus:sawdust (1:1) in greenhouse, the survival rate was more than 98%.
Shoot multiplication responses of three apple scions to different concentrations of BA and BAR as single source of cytokinins and in combination with two concentrations of KIN were studied. The effects of hormones depended on genotype, type and interactions of different cytokinins. Use of BAR significantly enhanced the shoot multiplication of c...v. Jonagold (6.5 shoots per explant). The multiplication rate of cv. Jonagold could not be improved by using the combination of BAR and KIN. The best proliferation was achieved by 1.0 mg 1-1 BA combined with 1.0 mg 1-1 KIN of cv. Prima..(8.1) and of cv. Galaxy (10.4).The effect of 0.5 mg 1-1 BA along with 1.5 mg 1-1 KIN was similar on multiplication rate (10.9) of cv. Galaxy.
The use of pathogen-free planting stock for new vineyard establishment is a key component in the maintenance and expansion of vine and quality table grape production. The success of the necessary changes in the structure of the grape industry is forced by the globalization process, the climate change, the rediscovery of autochton varieties as w...ell as breeding of new tolerant and resistant varieties. The renewal of vineyards largely depend on the availability of planting stocks. Serbia and Hungary found a common interest in establishing pathogen-free stock materials from newly breed resistant varieties and clonal selections of varieties which are traditional in the Serbian-Hungarian border area. During a cross-border cooperation program a complex system for the production of pathogen-free grapevine propagating material was established. Using heat therapy, in vitro shoot tip culture and traditional and molecular diagnostic techniques new pathogen-free stock materials were established from 26 varieties. They have been or will be tested for the presence of most important grapevine viruses, phytoplasmas, as well as bacterial and fungal pathogens. The complex system applying green grafting for indexing on grapevine indicators can shorten the duration of the procedure from 4 years to two-three years.
Factors 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%.
Callus 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, be...nzyladenine 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.
Hyperhydricity 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.
Shoot multiplication responses of rootstocks cvs. M.26, MM.106 and JTE-H to different concentration of BA, BAR and IBA in eight various combinations were tested on MS-medium. The effect of hormones depended on genotype, type of cytokinin and interaction of cytokinin and auxin. Shoot multiplication was significantly enhanced with the use of BAR...as cytokinin. High multiplication rate could be achieved in cvs. M.26, MM.106 and JTE-H: 7.7, 6.9 and 9.9 shoots per explant, respectively.
Some experience or details are introduced in connection with the nutrient uptake of micropropagated fruit trees in the different phase of the in vitro or ex vitro development. It can be stated, that the plants during the micropropagation procedure are overfed. Special careful nutrient supply is necessary during the acclimatiza...tion.
In vitro cultures have widely been used in horticulture for rapid multiplication of new varieties and clones as well as to produce pathogen-free stock material. To improve efficient hardening and transfer in vitro grown grapevine plants were multiplied by cutting them into single-node internodes with the whole leaf. Microcutti...ngs including the shoot tips were rooted in granulated perlite moisted with tapwater under sterile conditions. After 2-3 weeks the rooted microcuttings were supplied by nutrients and hardened by gradual opening and finally by complete removal of the lids of jars or plastic boxes used for growth. Using this method microcuttings of Vitis vinifera cvs. „Chardonnay", „Cabernet franc", „Riesling" and „Sauvignon blanc" and the rootstock varieties Vitis riparia x Vitis cinerea cv. „Barrier" and Vitis berlandieri x Vitis rupestris cv. „Richter 110" formed new roots and shoots and 100% of the tested plants survived the acclimatization procedure. Similar results were obtained when perlite was replaced with rockwool-, or pit-pot blocks. This method may highly increase the efficiency of producing pathogen-free propagating material and new transgenic lines.
In vitro plant material of clones (Q. robur) NL 100 A (adult) and NL 100 R (rejuvenated) received from Germany (A. Meier-Dinkel, 1995) were used in these experiments. WPM medium was used for the multiplication phase. Plantlets were subcultured monthly. Differences in quality and colour of the adult and rejuvenated cultures ind...uced us to follow and compare the changes of mineral- and chlorophyll content and dry weight during the propagation phase. Mineral and chlorophyll content as well as dry weight were measured weekly on three samples during the subculture period.
In the case of propagation rates we stated, they were similar around the year, but both clones had a high peak in April. Examining the cation-content, we detected that, the plantlets had a highest quantity of several elements during the 2nd and 3rd week of subculture. The iron content was the highest in the 1st week and after that it decreased continuously. It is supposed, that the content of iron is not enough in the media. The chlorophyll content of the rejuvenated clone was higher than that of the adult one.
In the rooting experiments it was stated that, after one-week cold treatment the rooting ability was the best.