The lignin contents and anatomical structure of roots of wild cherry (Prunus avium L.) and pedunculate oak (Quercus robur L.) plantlets were compared to explain differences in response during transfer from in vitro to ex vitro conditions. Lignification of cell walls increased significantly in both oak and cherry roots during the period of acclimation and finally lignin content of root tissues of in vitro propagated plantlets reached the levels not significantly different from seedlings grown in soil. Later on when secondary tissues appeared, lignified secondary xylem constituted most of the tissues of both species. The most conspicuous interspecific difference in root structure was the presence of phithickenings in cortical layers just outer to endodermis in cherry roots cultivated ex vitro. Formation of phi-thickenings was avoided in vitro and their presence thus seems to be under environmental control. Suberised well established exodermis was present in roots of oak but not detected in those of cherry. Very early development of exodermis in oak roots, preceding suberisation of endodermis, was recorded in vitro but not in well aerated soil. While multilayered and well-developed cork occurred in oak, only thin walled and less suberised secondary dermal tissues were found in cherry.
The polyamine (PA) contents and activities of PA biosynthetic enzymes in Norway spruce somatic embryos [Picea abies L. (Karst.), genotype AFO 541] were studied in relation to anatomical changes during their development, from proliferation to germination, and changes in these variables associated with the germination of mature somatic and zygotic embryos were compared. Activities of PA biosynthetic enzymes steadily increased during the development of somatic embryos, from embryogenic suspensor mass until early cotyledonary stages. In these stages, the spermidine (Spd) level was significantly higher than the putrescine (Put) level, and the increases coincided with the sharp increases in S-adenosylmethionine decarboxylase activity in the embryos. The biosynthetic enzyme activity subsequently declined in mature cotyledonary embryos, accompanied by sharp reductions in PA contents, especially in cellular Put contents in embryos from 6 weeks old through the desiccation phase (although the spermine level significantly increased during the desiccation phase), resulting in a shift in the Spd/Put ratio from ca. 2 in early cotyledonary embryos to around 10 after 3 weeks of desiccation. In mature zygotic embryos, Spd contents were twofold lower, but Put levels were higher, than in mature somatic embryos, hence their Spd/Put ratio was substantially lower (ca. 2, in both embryos and megagametophytes). In addition, the PA synthesis activity profiles in the embryos differed (ornithine decarboxylase and arginine decarboxylase activities predominating in mature somatic and zygotic embryos, respectively). The start of germination was associated with a rise in PA biosynthetic activity in the embryos of both origins, which was accompanied by a marked increase in Put contents in somatic embryos, resulting in the decline of Spd/Put ratio to about 2, similar to the ratio in mature and germinating zygotic embryos. The accumulation of high levels of PAs in somatic embryos may be causally linked to their lower germinability than in zygotic embryos.
Micropropagated plantlets derived from selected clones of the hybrid aspen (Populus tremula × Populus tremuloides) and the rowan-tree (Sorbus aucuparia L.) were used to determine the comparative study of uptake of the toxic, heavy metals Cd, Pb and the essential metal Mn. Samples of roots and aboveground parts (hypocotyl-derived tissues, leaves and stems) were taken from the plantlets grown for 24, 48, 96, or 168 hrs under aseptic conditions, in hydroponics with the toxic heavy metal and the essential metal salts. The concentration and distribution of the accumulated metals were determined using the ICP-OES method. The differences in the uptake capacity of hybrid aspen and rowan-tree clones for Cd, Pb and Mn were identified. Generally, the amounts of accumulated Cd and particularly Pb were much higher in the roots of both hybrid aspen and rowan-tree clones, than in their shoots, at all sample times. Conversely, the amounts of accumulated Mn were significantly lower than Cd and Pb in all plant parts of the hybrid aspen and rowan-tree samples. Patterns of Mn uptake were similar in the above-mentioned tissues of both clones, at all sample times. We concluded that the two clones of hybrid aspen and rowan-tree, lacking auxiliary soil microbiota, can accumulate large amounts of the toxic heavy metals Cd (800-1,500 mg/kg) and Pb (5,000-13,000 mg/kg) in roots and about 100 mg/kg of Cd was determined in aboveground part of hybrid aspen.
The study presents the comparative analyses of endogenous contents of auxin (IAA), cytokinins (CKs), polyamines (PAs), and phenolic acids (PhAs) in apical and basal parts of elm multiplicated shoots with regard to the organogenic potential. The shoot-forming capacity was higher in the apical part than in the basal part. However, the timing of root formation was in the apical type of explant significantly delayed (compared with the organogenic potential of basal part). Significantly higher contents of free bases, ribosides and ribotides of isopentenyl adenine, zeatin and dihydrozeatin that were found in the apical segments, might be considered as the most important factor affecting in vitro shoot formation. The content of endogenous free IAA was approximately three times higher in the basal shoot parts than in the apical parts. The amounts of putrescine and spermidine were higher in the apical part which generally contains less differentiated tissues than the basal part of shoot. The predominant PhA in both types of explants was caffeic acid, and concentrations of other PhAs decreased in the following order: p-coumaric, ferulic, sinapic, vanillic, chlorogenic, p-hydroxybenzoic and gallic acids. The contents of all determined PhAs in their free forms and higher contents of glycoside-bound p-coumaric, ferulic and sinapic acids, precursors for lignin biosynthesis, were found in the basal parts.
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