Adventitious rooting in microcuttings of Malus rootstocks MM106 was studied as regards their histological and biochemical aspects. Microcuttings from shoots raised in Murashige and Skoog's (1962) medium were transferred into a rooting medium containing IBA in the dark, then fixed 0, 3, 5, 7 and 10 days after. Some cambial zone and adjacent phloem cells became dense cytoplasm, nuclei with prominent nucleoli and the first cell divisions were observed at day 3. Meristemoids became individualized, consisting of densely staining cells (with enlarged nucleoli) formed outside the xylem by day 5. Identifiable root primordia with a conical shape and several cell layers were present at day 7. Roots with organized tissue system emerged from the stem 10 days after the root induction treatment. From these histological observations, it can be established that the rooting induction stage ended before day 3. The initiation stage, with the first histological modifications to the formation of meristemoids, would correspond to the transient increase of our biochemical marker (peroxidase activity) until day 5. The best rooting percentage obtained with cultures in the presence of auxin during 5 days confirms this hypothesis. The expression of rooting can then take place.
Gravity is a constant force guiding the direction of plant growth. In young poplar stem, reorientation of the apical region is mainly obtained by differential growth of elongating primary tissues. At the base, where elongation is achieved but where the cambium is active, reorientation is due to asymmetrical formation of reaction wood. After 45 min of gravistimulation, the stem showed no reorientation, but 1 week later, reaction wood was observed at the base of the stem. To determine the molecular mechanisms taking place at the top and base of the stem, after 45 min or 1 week of inclination, the changes induced in protein accumulation were studied by two-dimensional polyacrylamide gel electrophoresis and quantitatively analyzed using image analysis software. Around 300 protein spots were reproducibly detected and analyzed. Forty percent of these proteins showed significant changes after inclination. Mass spectrometry analysis of 135 spots led to the identification of 60 proteins involved in a wide range of activities and metabolisms. Very different patterns of protein expression were obtained according to conditions tested, highlighting the complexity of gravitropic responses. Our results suggest that primary and secondary tissues present specific mechanisms to sense reorientation and to respond to inclination. Some selected proteins are discussed.
Objective. Plant cell culture is an innovative technology to produce a variety of substances. Numerous plants synthesize among their secondary metabolites phenolic compounds which possess antioxidant and antimicrobial effects. Hawthorn (Crataegus) is one of these plants which has long been used in folk medicine and is widely utilized in pharmaceutical preparations mainly in neuro- and cardiosedative actions. Methods and Results. The production of polyphenol by fifty-two-week-old Crataegus azarolus var. aronia calli was studied in relation to growth variation and antioxidant and antimicrobial capacity within a subcultured period. The DPPH and ABTS+ assays were used to characterize the antioxidant actions of the callus cultures. Antimicrobial activity was tested by using disc diffusion and dilution assays for the determination of the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) values of each active extract. High TEACDPPH, TEACABTS, and antimicrobial activity was observed when maximal growth was reached. An optimum of total phenol, proanthocyanidins, flavonoid, (−)-epicatechin, procyanidin B2, chlorogenic acid, and hyperoside was produced during this period. Conclusion. Antioxidant and antimicrobial activities were strongly correlated with total phenols and total flavonoids. Crataegus azarolus var. aronia cells culture represents an important alternative source of natural antioxidants and antimicrobials.
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