Post-effects of cytokinins and auxin levels of proliferation media on rooting ability of in vitro apple shoots (Malus domestica Borkh.) 'Red Fuji'

Rooting ability of in vitro apple shoots of 'Red Fuji' grown on proliferation media with different hormone content were tested at three IBA levels in root induction media. Rooting percentage could be slightly increased with an increase in IBA concentration in proliferation media. The highest IBA concentration (3.0 mg 1-1) in root induction media showed strong inhibitory effect on rooting capacity of in vitro shoots. The highest rooting percentage (95%) could be achieved by shoots grown on proliferation media containing TOP or BA+KIN as cytokinins before rooting.


Knot formation by Pseudomonas syringae subsp. savastanoi on the in vitro shoots of Sorbus redliana

Two strains of Pseudomonas syringae subsp. savastanoi were isolated from Forsythia sp. and Nerium oleander in Hungary in 1997. The effects of growth regulators produced by the bacteria were studied in different experiments. The strains were co-cultured with Sorbus redliana in vitro shoots without being in contact with the plant on solid media. Further culture filtrates in different concentrations were added to the culture medium. The growth regulators presented in the agar caused knot formation on the shoots and on the leaves in both kinds of culture. There were significant differences in the cultural and physiological characters, auxin and cytokinin activity of the strains of different origin.


Effect of reduced spray programmes on incidences of apple scab, powdery mildew and codling moth damage in environmentally freindly apple production systems

Still a large amount of pesticides and spary applications are used in environmentally friendly fruit production systems; therefore, the aim of our study was first to test the in vitro effeicacy of some fungicides against a key apple disease (apple scab), and secondly to evaulate the effectiveness of reduced spray programmes against apple scab, powdery mildew and coding moth in integrated and organic apple orchards. In vitro efficacy of 7 fungicides (Champion 50 WP, Kocide 2000, Nordox 75 WG, Olajos rézkén, Kumulus S, Rézkén, Rézoxiklorid) and another 6 fungicides (Score 25 EC, Efuzin 500 SC, Systane, Folicur Solo, Zato Plusz, Rovral) approved in organic and integrated production systems, respectively, were tested against apple scab. Altogether four spray programmes were performed i) standard integrated: sprays followed by forecasting systems during the season, ii) reduced integrated: sprays followed by forecasting systems but only 75% of the spray numbers used during the season-long spray programme, iii) standard oragnic: sprays applied every 7–14 days during the season and iv) reduced organic: 60% of the spray numbers used during the season-long spray programme. In vitro results showed that fungicides (with active ingredients of copper and sulphur) applied in organic production showed relatively high percent growth capacity of the apple scab fungus. Rézkén showed the highest and Kumilus S the lowest efficacy against apple scab. Fungicides applied in integrated production showed relatively low percent growth capacity of the apple scab fungus. Score 25 EC showed the highest and Rovral the lowest efficacy against apple scab. Field study showed that reduced spray programmes did not increase significantly scab incidence in the integrated field. However, scab incidence increased significanly (above 30%) in the reduced spray programme for the organic orchard.Mildew incidence was low (below 5%) in both integrated and organic spray programmes. Mildew incidence on both shoots and fruits increased significanly in the reduced spray programme for the organic orchard. Incidence of codling moth damage was affected the most by standard vs. reduced spray programmes. Though incidence remained below 10% in the integrated plots, the incidence was significantly higher in the reduced spray programme compared to the standard programme. Similar results were obtained for organic spary programmes, but the incidence was 10 times higher and the differences among the two programmes were larger.,

In vitro propagation of 'Echo' cultivars of Eustoma grandiflorum (Raf.) Shinn.

Eustoma grandiflorum (Raf.) Shinn. 'Echo' Fl cultivars ('Echo White', 'Echo Rose', 'Echo Blue', 'Echo Blue Picotee') were used and multiplication of shoots was evaluated on Murashige and Skoog (1962) basal medium with 11 g/1 agar-agar and 20 g/1 sucrose. To test the effect of BA different concentrations were added: 0.10, 0.25 mg/1 and a culture medium without BA. Differentiation of roots was examined on Jámbor-Benczúr and Marta (1990) basal medium with the same concentration of agar-agar and sucrose. To examine the effect on rooting, various concentrations of NAA were used: 0.5, 1.0, 2.0, 3.0 mg/l. The pH was adjusted to 5.6 in every case using KOH. We studied the after-effect of different concentrations of BA during the acclimatisation. During the multiplication, the cultivar 'Echo White' formed the most shoots and the smallest leaves on the medium with 0.10 mg/1 BA. Fortunately, in the case of this cultivar, the number of shoots was reduced and the length of leaves was increased succesfully on the medium without BA. The other three cultivars developed the longest leaves on the medium containing 0.10 mg/1 BA. Sometimes not only shoot regeneration but spontaneous rooting was observed during the multiplication. Examining the rooting, the highest percent of roots was found on the medium with 1.0 mg/1 NAA, and the cultivar 'Echo Rose' formed the most roots on this medium. Higher concentration (2.0 and 3.0 mg/1) of NAA already reduced the number of roots in all of the cultivars. During the acclimatisation, the percentage of survival was 76.3% and the tallest plants with the longest leaves were found on the multiplication medium with 0.25 mg/1 BA. 'Echo Blue Picotee' gave the best results with the tallest pieces and longest leaves on this medium.

How to produce large sized microtubers of potato cv. Desiree

In vitro tuberization was induced on explants with different number of nodes layered on a medium with high sucrose (8%) content: 30, 15, 10, 7 and 6 explants per jar were cultured containing 1, 2, 3, 4 or 5 nodes, respectively. Microtubers developed were graded by their smallest diameter, and the number of tubers per jar, their size distribution, their fresh weight and the multiplication rate were recorded. The highest multiplication rate (1.98) was obtained for explants with 5 nodes. The size distribution of tubers was markedly affected by treatments. The majority of microtubers (49.4%) were 6-8 mm in the case of the smallest explants (with I node). When explants with 2 to 5 nodes were used, the most microtubers were 8-10 mm but with an increase of explant size, more and more microtubers were produced with larger diameter up to 16 mm and average fresh weight of tubers also increased with the increase of explant size. For the microtuber production of Desiree the use of explants with two nodes can be suggested because in this treatment the average fresh weight of microtubers was high enough (250 mg) and the number of large sized microtubers was very high (79% was larger than 6 mm and 53% was larger than 8 mm).

Inhibition and recovery of germination and growing ability of seedlings under and after osmotic stress induced by polyethylene glycol in 8 pea genotypes

Germination and early seedling growth of eight pea genotypes were examined under and soon after different (5%, 10%, 15%, and 20%) PEG treatments. Seeds were germinated on PEG solution for 3 or 6 days and then further germinated and cultured on filter paper moistened by water for 3 and 6 days. The length and fresh weight of shoots and roots of seedlings were measured and used for evaluation of genotypes. Roots were less inhibited by osmotic stress than shoots similarly to other plant species. The variability among the genotypes was the greatest in the case of shoot growth at 5% PEG treatment and in the case of root growth at 15% PEG treatment. Results suggest that growing responses of genotypes after cessation of stress are more suitable for the evaluation of their osmotic tolerance, than their responses expressed during in vitro stress conditions. Genotypes with relatively high or low osmotic stress tolerance, respectively, could be distinguished with 6 days after recovery from 3-day-long 15% PEG treatment concerning the rate of shoot weight to root weight.

Examinations of potential environmental friendly materials against tomato and pepper patogens

In organic farming systems the focus is on prevention with regards to plant protection. To follow the rules of Good Agricultural Practice one is able to avoid serious yield losses; if it is not possible the use of allowed materials are permitted. Organic farmers have less material to protect their plants so it is necessary to find effective potential materials. Bacterial and fungal diseases of tomato and pepper can cause serious losses in yield. Different materials were tested against some plant pathogen bacterial (Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, Xanthomonas campestris pv. vesicatoria and fungal (Phytoptora infestans, Rhizoctonia solani, Sclerotinia sclerotium) strains in order to find potential materials in the field of organic seed treatment. In vitro trials have shown that vinegar, cider vinegar, red wine vinegar, white wine vinegar, cinnamon and thyme oil have inhibiting effect against the causative agent of bacteria and fungi. Germination test has shown that examined vinegar types do not decrease germination ability if the concentration is low, but in higher (more than 5%) concentration it ruins the germination ability. Even in 0,5% concentration of red — and white vine vinegar have good effect on germination capacity.

Morphological, physiological features and differences of Vriesea splendens ’Fire’ plants during in vitro multiplication and rooting

During in vitro multiplication and rooting of Vriesea splendens ’Fire’, 0.1, 0.2, 0.4 and 0.8 mg l-1 benzyladenine (BAP), benzyladenine-riboside (BAPR), kinetin (KIN), meta-topoline (MT), indole-butyric acid (IBA) and naphthalene-acetic acid (NAA) were added to basal Murashige and Skoog (1962) MS medium. As compared to the hormone-free control, plants developed significantly more shoots on medium supplemented with almost all cytokinins (excepting KIN), especially BAP resulted the highest multiplication up to almost 26 shoots. Enhancement of cytokinin concentrations increased shoot number (and in case of BAP, peroxidase activity) but decreased plant height and rooting parameters. Regarding root production, both auxins were definitely beneficial (0.2 mg l-1 NAA resulted more than 7.5 roots and higher auxin concentrations efficiently stimulate root elongation); however, KIN had similar effects. After a three-month duration time of acclimatization, we observed that plants which were previously cultured on medium containing certain cytokinins (KIN in all doses and 0.1 mg l-1 MT) or both auxins had greater survival, moreover, as negative after-effect, higher cytokinin concentrations reduced the number of survived specimens.

Results in the vitro propagation acclimatization of Pontederia lanceolata

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 Pontederia lanceolata.


The effect of different cytokinins on chlorophyll content and morphological features of in vitro Nidularium ’Kertész Jubileum’

During in vitro multiplication of Nidularium ‘Kertész Jubileum’, 20 g/l sucrose, 5 g/l agar, 100 mg/l inositol, and different concentrations of benzyladenine (BA), benzyladenine-riboside (BAR), kinetin (KIN), meta-topolin (mT) were added to the MKC (Knudson, 1946) basal medium. Furthermore, 0.1 mg/l naphthaleneacetic acid was used to every medium. Number of shoots, length of leaves, number and length of roots, chlorophyll (a+b) content were examined and evaluated with Ropstat statistical software. As compared to the other cytokinin, significantly most shoots were obtained in the case of applying BA. Increasing of BA-concentration (as far as 2 mg/l) enhanced shoot number (from 10.92 to 19.26) but 4 mg/l BA resulted only 6.63 shoot. The less efficient cytokinin was KIN, in most cases no more than about 2 shoot was achieved. Regarding the length of leaves, the higher level of BA effected averagely the shorter leaves (from 24,46 to 7.31 mm). KIN effected significantly the longest leaves (43.4-61.29) in inverse proportion to the concentration. The same cytokinin resulted the most (and the longest) roots with the highest rooting percentages, but more KIN decreased the number and length of roots (from 7.95 to 4.4 and from 38.49 to 22.73 mm). There were no definite correlation between cytokinin concentration and chlorophyll (a+b) content, but the highest doses resulted decreasing (except of meta-topolin which leads to the lowest values). Summarizing, BAR effected the highest contents (mostly more than 1400 μg/g), particularly in the case of 1 mg/l (1807.3 μg/g).

Aminoglycoside antibiotics affect the in vitro morphogenic response of chrysanthemum and tobacco

Broadly the success of genetic transformation of plants requires non-chimeric selection of transformed tissues and its subsequent regeneration. With rare exceptions, most plant transformation protocols still heavily utilize antibiotics for the selection of transgenic cells containing an antibiotic-degrading selectable marker gene. The morphogenic capacity of in vitro chrysanthemum and tobacco stem and leaf explants change with the addition of aminoglycoside antibiotics (AAs). Of 6 antibiotics tested, phytotoxicity occurred at 10-25 and 50-100 pgml-I in chrysanthemum and tobacco explants, respectively, depending on the size of the explant and the timing of application. The presence of light or darkness also had a significant effect. The use of transverse thin cell layers (tTCLs) in conjunction with high initial AA selection levels supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow cytometric analyses demonstrate that regeneration can be predicted in both species, depending on the ploidy level of the callus. Endoreduplication was not observed in chrysanthemum, even at high AA levels, but occurred (8C or more) in tobacco callus, even at low AA concentrations (5-10 pgml-1). The higher the AA level, the greater the DNA degradation and the lower the 2C and 4C values.

Factors affecting rhizogenesis in vitro and acclimatisation of three cherry rootstocks

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%.

Effects of aromatic cytokinins on structural characteristics of leaves and their post- effects on subsequent shoot regeneration from in vitro apple leaves of 'Royal Gala'

The effects of different aromatic cytokinins applied in different concentrations and combinations were investigated on the histology of in vitro apple leaves and their post-effects on subsequent shoot regeneration from these leaves were studied. Great differences in the anatomical structure of leaves could be detected originating from media containing different types and concentrations of aromatic cytokinins. The number of regenerated shoots per explant and the organogenetic index were used for the evaluation of the post-effect of aromatic cytokinins on shoot regeneration. The histological structure of leaves used for regeneration and their regeneration response showed a good correlation. When the pre-treatment caused a juvenile-like or less-differentiated structure, the number of regenerated shoots per explant increased and often vitrification also decreased and consequently the organogenetic index also increased. A strong interaction between cytokinin-content (type and concentration) of the pre-treatment medium and that of the regeneration medium could also be detected.

In vitro plant regeneration from immature embryo axis and cotyledons of common bean (Phaseolus vulgaris L.)

Phaseolus 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.

Influence of different growth regulators on the in vitro morphogenesis of an ornamental variety of carnation

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, 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.

Dr. Ottó Orsós, the forgotten Hungarian pioneer in plant tissue culture

The 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.

The role of rejuvenated and adult forms of English oak (Quercus robur) in in vitro cultures

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 induced 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.


In vitro investigation of King Oyster Mushroom [Pleurotus eryngii (DC.: Fr.) Quel.] strains in vegetative growing phases

The king oyster mushroom ( Pleurotus eryngii) is more and more popular amongst the producers due to its excellent taste and relatively easy cultivation . In the course of our work we collected 15 king oyster mushroom strains from various habitats in Hungary in order to get a better picture about the growth of the vegetative mycelia of the species and its different strains. In the in vitro experiments we investigated the growth of the strains at various temperatures and pH. incubated them in light and darkness and in aerobic and anaerobic atmosphere. In addition to these we measured the weight of dry mycelia produced in a given time by the strains. Our results showed that the above mentioned environmental conditions resulted in a very different growth rate of the vegetative mycelia of the various P. eryngii strains. These results may provide valuable data about the vegetative phase of the cultivation.

Effect of gelling agents on in vitro development of Amelanchier canadensis ‘Rainbow Pillar’

In vitro shoot multiplication responses of Amelanchier canadensis ‘Rainbow Pillar’ were studied on media solidifi ed with different gelling agents. The media were gelled either with 6.8 g l-1 fi brous agar-agar, or 50.0 g l-1 wheat starch, or 20.0 g l-1 Guar gum, or 15 g l-1 Isubgol or 50.0 g l-1 wheat starch mixed with 0.5 g l-1 Phytagel. Shoot cultures were grown for two months, thereafter the multiplication rates (number of newly developed shoots per explant) were counted and the length of shoots were measured. We found that the highest shoot multiplication of Amelanchier canadensis ‘Rainbow Pillar’ occurred on media gelled with Guar gum, while the longest shoots developed on media with Starch. About four-fold shoot number were obtained on media with Guar gum compared to the weakest results found on media gelled with Isubgol. Finally, considering all factors (shoot growth parameters, costs) the most economical gelling agent for Amelanchier canadensis ‘Rainbow Pillar’ was proved to be wheat starch among the tested alternatives which allows a 75.6% cost reduction.

Effects of tuberization conditions on the microtuber yield and on the proportion of microtuber tissues

The 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.

Micropropagation of Rudbeckia hirta L. from seedling explants

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 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.

Effect of aromatic cytokinins and explant position on shoot multiplication of Asparagus offi cinalis L.

Asparagus offi cinalis has been widely studied, but little information is available about its in vitro response to exogenous cytokinin during shoot multiplication. To study the effects of different cytokinins on shoot multiplication of A. offi cinalis ‘Grolim’, in vitro culture was initiated from shoot segments cultured on media with Murashige and Skoog medium. Effects of different aromatic cytokinins (6-benzylaminopurine, 6-benzylaminopurine riboside and meta-topolin) applied in four concentrations (0.5, 1.0, 1.5, 2.0 mg/l) on shoot multiplication of ‘Grolim’ were tested. Effect of explant position (vertically or horizontally) on the shoot multiplication outcome was also studied. Both the length and the number of newly developed shoots were signifi cantly affected by explant position and cytokinin content of the medium. The highest numbers of shoots (4.9) were produced in the presence of 0.5 mg l-1 6-benzylaminopurine riboside when explants were paced horizontally onto the medium. Although the longest shoots (41.5 mm) developed on explants placed vertically onto medium supplemented with 2.0 mg l-1 meta-topolin, the lengths of shoots developed on medium with 0.5 mg l-1 6-benzylaminopurine riboside were also adequate in both explant position (29.5 and 33.6 mm placed horizontally and vertically, respectively).

In vitro comparative study of two Arundo donax L. ecotypes’ selenium tolerance

Selenium tolerance of two somatic embryo-derived Arundo donax L. ecotypes (Blossom, 20SZ) were compared in in vitro culture. Sodium-selenate (1 – 100 mg L-1) as known the most phytoavailable selenium form and the less studied red elemental nanoselenium (100 mg L-1) were applied as selenium treatments. Basis on the results Blossom ecotype seemed to be more sensitive to the sodium-selenate than 20SZ. Inhibiting effect of selenate was effectuated above 10 mg L-1 in case of Blossom, which was manifested in decreased survival rate and growing parameters. Contrast to this 20SZ could tolerate the selenate ≤ 20 mg L-1 without any toxic symptoms. Lower selenate tolerance of Blossom could be explained with higher selenium accumulation. Both of two ecotypes could also uptake and accumulate the red elemental nanoselenium however in much less extent compared to selenate.

Effects of activated charcoal on rooting of in vitro apple (Malus domestics Borkh.) shoots

Rooting of in vitro 'Royal Gala' shoots was studied under different conditions of root induction and root elongation phase. The rooting capacity was affected by both rooting phases. Very high rooting percentage could be reached with both liquid and solid root induction media. Raising the temperature from 22 °C to 26 °C during root induction phase increased the rooting percentage. Presence of activated charcoal in root elongation media can affect the number of roots per rooted shoots and can increase the rooting percentage, the length of roots and the rate of survival depending also on other conditions during rooting. Presence of NAA in root elongation media reduced the number and the length of roots considerably. Favourable effect of activated charcoal on rooting was mainly due to adsorption of NAA.

Application of the Jerusalem artichoke (Helianthus tuberosus L.), as a plant origin medium additive, during the micropropogation of Ada keiliana

A procedure for in vitro propagation of Ada keiliana seedlings are suited for acclimatization, was worked out. M medium was supplemented, with Jerusalem artichoke, as plant origin complex additive. The apply of JAD (1,5g/flask) gave the best response, considering the shoot (29 mm), and the root development (24,9) mm) too. The plantlets with satisfying growth (25-30 mm, 4-5 roots) were transferred in small pine bark: Novobalt peat: coconut fibres: perlit (2:3:1:1) mix, among greenhouse circumstances.

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