The effect of different biostimulators on morphological and biochemical parameters of micropropagated Hosta ’Gold Drop’
22-29.Vol 25No 1-22019
During 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.
131-136.Vol 20No 3-42014
Jerusalem 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.
81-83.Vol 5No 1-21999
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.
123-129.Vol 20No 3-42014
Explants 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 a...chieved 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.
61-69.Vol 14No 32008
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.
In vitro effect of different cytokinin types (BAP, TDZ) on two different Ocimum basilicum cultivars explants
15-20.Vol 25No 3-42019
Ocimum basilicum L. (sweet basil) is an economically and ethnobotanically important aromatic, medicinal, ornamental and culinary herb, with a very wide gene pool, that is sensitive to cold and prone to several plant pathogens that can demolish harvest and lessen yield. In this research, the effects of BAP (6-Benzylaminopurine) and TDZ (Thidiazuron...) on different genotypes for in vitro cloning were determined, in order to provide a detailed protocol guide concerning Ocimum basilicum L. propagation. The results from the O. basilicum seed propagations revealed that the best condition for the secondary shoot growth is with 5.0 mg/l TDZ or 1.5 mg/l BAP on all types of explants except the root, the secondary root growth can be obtained on all types explant with any BAP concentration and all cytokinins can induce callus on all types of explants. On the whole, it shows that multiple secondary shoot induction and regeneration in Ocimum basilicum L. is regulated by appropriate cytokinin concentration.
47-49.Vol 6No 12000
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 lanceol...ata.
61-64.Vol 13No 12007
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 differentiation 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%.
105-108Vol 11No 22005
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 an...d, 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.
Application of the Jerusalem artichoke (Helianthus tuberosus L.), as a plant origin medium additive, during the micropropogation of Ada keiliana
61-64.Vol 14No 42008
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 satisf...ying 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.
87-91.Vol 11No 22005
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 me...dium 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.
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