RESUMEN
In plants, phase change from the juvenile stage to maturity involves physiological and anatomical changes, which are initiated and controlled by evolutionary highly conserved microRNAs. This process is of particular significance for the in vitro propagation of woody plant species, as individuals or tissues that have undergone the transition to vegetative maturity are recalcitrant to propagation. Conserved miRNAs differentially expressed between juvenile (including rejuvenated) and mature silver birch tissues were identified using high-throughput sequencing of small RNA libraries. Expression of some miR156 isoforms was high in juvenile tissues and has been previously reported to regulate phase transitions in a range of species. Additional miRNAs, such as miR394 and miR396, that were previously reported to be highly expressed in juvenile woody plant tissues were also differentially expressed in this study. However, expression of miR172, previously reported to be highly expressed in mature tissues, was low in all sample types in this study. The obtained results will provide insight for further investigation of the molecular mechanisms regulating vegetative phase change in silver birch and other perennial woody plant species, by analysing a wider range of genotypes, tissue types and maturation stages. This knowledge can potentially assist in identification of rejuvenated material at an earlier stage than currently possible, increasing the efficiency of silver birch in vitro propagation.
RESUMEN
Saussurea esthonica is an endangered plant species typical for wet inland habitats such as calcareous fens. Due to its limited population size and distribution, non-invasive sampling of is important in the research of S. esthonica. The aim of the present study was to assess the effect of mineral nutrient availability and substrate moisture on the growth, physiological status, and mineral nutrition of S. esthonica. The non-destructive measurement of physiological parameters was performed in native habitats during three vegetative seasons, followed by two experiments in controlled conditions. Soil at the two Estonian sites had a relatively larger similarity in the composition of plant-available mineral nutrients in comparison to the two Latvian sites. The chlorophyll a fluorescence parameter Performance Index correlated with the total precipitation in the respective month before measurement, but no significant relationship with other environmental variables was found. For mineral nutrient experiments, plants were grown in four substrates with different mineral nutrient composition, resembling that of soil at different S. esthonica sites. Plant growth and physiological indices were significantly affected by the mineral composition of the substrate. Differences in leaf and root mineral nutrient concentrations of S. esthonica plants in part reflected differences in substrate mineral concentration. To evaluate the effect of soil moisture on growth and photosynthesis-associated parameters of S. esthonica, plants were cultivated in "Pope+" substrate at four different moisture treatments (dry, normal, wet, and waterlodged). The most intense growth of S. esthonica plants was evident in waterlodged conditions, which decreased with a decrease in soil moisture. The biomass of leaves increased by 106% and that of the roots increased by 72% as soil moisture increased from dry to normal. For waterlodged plants, leaf biomass increased by 263% and root biomass increased by 566%, in comparison to that for plants cultivated in dry substrate. Substrate drying had a more negative effect on the growth of S. esthonica plants in comparison to that of waterlodging, and this can be directly linked to prevalent hydrological conditions of an alkaline fen habitat native to the species. Therefore, the preservation of the natural water regime in natural habitats is critical to the conservation of the species.
RESUMEN
Micropropagation of forest reproductive material is becoming an increasingly important tool of climate-smart forest management, whose efficiency is depending on artificial illumination, which in turn can have species-specific effects. To improve the energy-efficiency of micropropagation, light emitting diodes (LED) are becoming more popular; however, they emit light of narrow spectral composition, synergic effects of which can alter plantlet development. Regarding the in vitro cultures of trees, such effects have been scarcely studied. In this study, three clones of silver birch (Betula pendula Roth.) and three clones of hybrid aspen (Populus tremuloides Michx. × Populus tremula L.) from the eastern Baltic region were tested. The responses of leaf and stem anatomy of in vitro cultures to three LED light illumination treatments differing by spectral composition and to illumination by fluorescent tubes were estimated by linear (mixed) models. The studied light treatments had non-interacted effects on stomata density and on the secondary xylem cell wall in the stem of silver birch and in the stomata length, stem radius, and phloem width of hybrid aspen. Furthermore, clone-specific responses to illumination were observed for number of chloroplasts and phloem width of silver birch and for leaf thickness and xylem cell wall thickness of hybrid aspen, implying different mechanisms of shade avoidance. In general, the responses of plantlet anatomy differed according to the width of the light spectrum in case of LED, as well as for fluorescent tubes. Considering the legacy effects of early development of plantlets, adaptability of illumination in terms of spectral composition according to the requirements of genotypes appear highly beneficial for micropropagation of sustainable forest reproductive material.
RESUMEN
The aim of the present study was to establish an experimental system in controlled conditions to study the physiological effect of abiotic/biotic interaction using a rare wild leguminous plant species from coastal sand dunes, Anthyllis maritima. The particular hypothesis tested was that there is an interaction between sand burial, rhizobial symbiosis and salt treatment at the level of physiological responses. Experiment in controlled conditions included 18 treatment combinations of experimental factors, with two intensities of sand burial, rhizobial inoculation and two types of NaCl treatment (soil irrigation and foliar spray). Shoot biomass was significantly affected both by burial and by inoculation, and by interaction between burial and NaCl in the case of shoot dry mass. For plants sprayed with NaCl, burial had a strong significant positive effect on shoot growth irrespective of inoculation. General effect of inoculation with rhizobia on shoot growth of plants without NaCl treatment was negative except for the plants buried 2 cm with sand, where significant stimulation of shoot dry mass by inoculant was found. The positive effect of burial on shoot growth was mainly associated with an increase in leaf petiole height and number of leaves. Performance index significantly increased in buried plants in all treatment combinations, and leaf chlorophyll concentration increased in buried plants independently on burial depth, and only in plants not treated with NaCl. Inoculation led to significant increase of leaf peroxidase activity in all treatment combinations except NaCl-irrigated plants buried for 2 cm by sand. Sand burial stimulated peroxidase activity, mostly in non-inoculated plants, as inoculation itself led to increased enzyme activity. In conclusion, strong interaction between sand burial and NaCl treatment was evident, as the latter significantly affected the effect of burial on growth and physiological indices. Moreover, rhizobial symbiosis had a significant effect on physiological processes through interaction with both sand burial and NaCl treatment, but the effect was rather controversial; it was positive for photosynthesis-related parameters but negative for growth and tissue integrity indices.
RESUMEN
The aim of the present study was to analyze the growth and physiological performance of two coexisting species, Trifolium fragiferum, and Trifolium repens, under the effect of NaCl and rhizobial symbiosis. Seeds of T. fragiferum and T. repens were collected from populations in the wild, and plants were cultivated in an automated greenhouse, two plants per container. Three basic types of planting were performed: (1) both plants were T. fragiferum (single species), (2) one T. fragiferum and one T. repens (species coexistence), (3) both plants were T. repens (single species). For every basic type, three subtypes were made: (1) non-inoculated, (2) inoculated with rhizobia taken from T. fargiferum, (3) inoculated with rhizobia taken from T. repens. For every subtype, half of the containers were used as control, and half were treated with NaCl. Shoot fresh mass of plants was significantly (p < 0.001) affected by species coexistence, inoculant, and NaCl. Three significant two-way interactions on plant shoot growth were found: between species coexistence and NaCl (p < 0.001), inoculant and species (p < 0.05), and NaCl and species (p < 0.001). A significant three-way interaction between inoculant, NaCl, and species (p < 0.001) indicated different responses of shoot growth of the two species to inoculant type and NaCl. NaCl treatment was an important factor for T. fragiferum, resulting in better growth in conditions of species coexistence, but the positive effect of bacterial inoculant was significantly more pronounced. A decrease in peroxidase activity in leaves was a good indicator of relative NaCl tolerance, while the absence/presence of rhizobial inoculation was reflected by changes in leaf chlorophyll concentration and photochemical activity of photosystem II. It can be concluded that interaction between biotic and abiotic factors affected the outcome of the coexistence of the two Trifolium species. Distribution of T. fragiferum in sea-affected habitats seems to be related to a higher competitive ability with allied species at increased substrate salinity, based on better physiological salinity tolerance.
