Model experiments for establishment of in vitro culture by micrografting in apple47-49.Views:167
Micrografting was used in our experiments for establishment of in vitro culture from one rootstock (`JTE-F') and three scion cultivars (`Remo', 'Rewena' and `Reanda') of apple. Shoot tips of these cultivars were harvested from field and grafted onto in vitro rootstock cultivars. Their survival and development were studied. 42-93% of shoot tips survived and developed further depending on cultivar. Impermanent browning of sticking agar-agar could be observed in 21-25% of the micrografts depending on cultivars but discolouration of agar-agar ceased within one week and did not cause any death of shoot tips. We used micrografting successfully for establishment of in vitro culture from cultivars, from which earlier with conventional methods the culture establishment was not possible because of hard tissue browning. However, further studies are necessary to ensure the survival and development of shoots after removing them from micrografts.
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'41-46.Views:147
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.
Effects of different cytokinins on the shoot regeneration from apple leaves of 'Royal Gala' and 'M.26'69-75.Views:187
The effects of different types of cytokinins on the shoot regeneration from leaf explants of apple scion 'Royal Gala' and apple rootstock 'M.26' were evaluated. Regeneration media contained either thidiazuron, or 6-benzylaminopurine, or meta-topolin, or zeatin, or kinetin, or their N9-ribosides, respectively, in the concentration range 0.5 to 8.0 mg 1-1. Effects of 'these cytokinins were evaluated on the percentage of regeneration (R%) and that of vitrification (V%) and on the number of regenerated shoots per explant (SN). Organogenetic index (0I) calculated from these data was used for the evaluation of efficacy of cytokinins. The course of shoot organogenesis also was followed using stereomicroscope. Types and concentrations of cytokinins applied in the regeneration media influenced each parameter significantly and the regeneration answer was strongly genotype-dependent. The best regeneration (SN: 11.08, 01: 7.5) was achieved in `Royal Gala' by using TDZ in concentration of 0.5 mg 1-1 (2.271,1M). There was a clear relationship between the effect on the regeneration efficacy and the chemical structure of cytokinins considering classical cytokinins, namely N9-ribosides applied in less concentration than nonribosides have the same or best regeneration effects except for 6-benzylaminopurine riboside. However, similar relationship could not be detected in the case of 'M.26'. SN was the highest (3.22) using 6.5 mg 1-1 (18.2011M) 6-benzylaminopurine riboside or 8.0 mg 1-1 (21.44 µM) meta-topolin riboside (3.18). SN was not significantly lower (3.12) by using 2.0 mg 1-1 (9.08 1M) TDZ, however, OI was about half as big (0.63 compared to 1.29 or 1.74 with 6-benzylaminopurine riboside or meta-topolin riboside, respectively). 'Royal Gala' had higher organogenetic ability, than `M.26': 3.5-fold higher shoot number per explant and more than 4-fold higher organogenetic index was reached with this cultivar than with 'M.26'. Moreover, the similar developmental stage of shoots could be observed 3-5 days earlier than in 'M.26' and if explants of 'Royal Gala' were further cultured with 3 weeks, SN increased from 11.08 to 24.42 on TDZ-containing regeneration medium, which might suggest higher organogenetic ability, too.
Effect of gelling agents on in vitro development of Amelanchier canadensis ‘Rainbow Pillar’75-79.Views:226
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.
Effect of aromatic cytokinins and explant position on shoot multiplication of Asparagus offi cinalis L.67-70.Views:222
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).