Indirect shoot organogenesis and plantlets regeneration from stem of ornamental Dieffenbachia maculata cv. Marianna (original) (raw)
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Indole-3-acetic acid (IAA) in the supernatant of a culture from the strains, Rhizobium leguminosarum biovar viciae, Bradyrhizobium japonicum, Pseudomonas sp. and Azotobacter sp was detected. Azotobacter sp yielded the highest concentrations of IAA. It was shown that the indole-3-acetic acid (IAA) was induced by the presence of tryptophan, which is used as inducer because the plant provide the bacteria with tryptophan under natural conditions. The highest concentration of IAA was produced by Azotobacter sp.(A1) at the end of the logarithmic phase (after 3days). The results obtained in this work provide useful information about the production behavior of IAA under the optimal conditions(temperature 30 degrees C and pH 7) which is of importance for the application in production Dieffenbachia maculat cv. Marianne plants by using tissue culture technique. This work was also conducted to study the effect of some growth regulators such as 10 mg/l IAA (synthetic), 5 mg/l BA and 10 mg/l IAA (bioauxin) on callus formation of Dieffenbachia maculat cv. Marianne shoot tips and internodal segments were taken from sterilized shoot and cultured on MS medium supplemented with 6 different treatments from growth regulators. Explants cultured on MS medium supplemented with either 10 mg/l IAA +5 mg/l BA or 10 mg/l bioauxin + 5 mg/l BA had the highest callus percentage 97.22 and 93.94%, respectively. MS medium supplemented with 2 mg/l BA + 0.06 mg/l BA was used for callus differentiation.
Recent Advances in the Application of Plant Tissue Culture in Dieffenbachia
Plant tissue culture has been shown to be a very important tool for the ornamental foliage plant industry. This is especially true for the foliage plant genus Dieffenbachia. The application of in vitro culture of Dieffenbachia has the potential to overcome some of the limitations associated with traditional methods of mass propagation, breeding and genetic manipulation. However, compared to other species, this approach has been limited in Dieffenbachia due to its recalcitrant nature in vitro. Recent advances in the application of plant tissue culture methods for the propagation and genetic manipulation of Dieffenbachia varieties are reviewed.
An efficient shoot regeneration protocol based on callus induction and multiple shoot development has been standardized for Plectranthus barbatus. The leaf explants were cultured on Murashige and Skoog (MS) [1] medium supplemented with α-naphthalene acetic acid (NAA at 2.68 – 16.11 μM) alone or in combination with Kinetin (KIN) and 6-benzyladenine (BA) for callus induction. The best callus production was obtained with medium containing NAA 10.74 μM with BA 2.66 μM. After two weeks of growth calli were transferred to shooting medium containing BA 11.10 μM and NAA 3.22 μM, shoot regenerated with a frequency of approximately 28.21 ± 3.10 shoots per callus and maximum average shoot length (8.37 ± 2.03 cm) were recorded on half strength MS medium supplemented with 1.73 μM Gibberellic acid (GA3) and 2.32 μM KIN. Rooting was best achieved on half-strength MS medium augmented with 7.38 μM Indole-3-butyric acid (IBA). The plantlets regen-erated in vitro with well developed shoot and roots were successfully established in pots containing garden soil and grown in a greenhouse with 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation. The described method can be successfully employed for large-scale multiplication and long term in vitro conservation of Plectranthus barbatus.
Callus Induction and Plantlet Regeneration in Soymida febrifuga A. Juss (Roxb.)
In vitro seedling explants of Soymida febrifuga were screened for their callus induction and morphogenic potential by culturing on Murashige and Skoog (MS) medium supplemented with naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), indole3acetic acid (IAA) alone or in combination with benzyladenine (BA), kinetin (KN), 2-iP (2-isopentynyl adenine), coconut milk. Multiple shoots were regenerated from cotyledonary node, shoot tip explants derived callus by one stage process where caulogenesis occurred on the primary callus. However cotyledons, leaves and root segments gave rise to adventitious shoots from callus by two stage method of indirect organogenesis. High frequency of shoot differentiation was observed from cotyledon and leaf on MS medium fortified with BA (8.9 to 13.3 μM) in combination with NAA (0.54 μM) compared to other explants. Further, when elongated shoots were subjected to rooting, we observed shoot necrosis consistent to our previous findings. Similarly, we could overcome this problem by supplementing calcium to MS rooting medium. In vitro rooted plantlets were successfully transferred to soil, exhibiting a normal development. Altogether, our protocol provides a rapid technique for mass multiplication as well for ex situ conservation of this endemic medicinal plant.