Photobiological and lipidic responses reveal the drought tolerance of Aster tripolium cultivated under severe and moderate drought: Perspectives for arid agriculture in the mediterranean.

In the past Aster tripolium has already proved to be a good candidate for saline agriculture in soils with low water availability. Thus, the aim of the present work was to disentangle the photobiological and biochemical mechanisms underlying the response of A. tripolium to PEG-induced drought stress, by exposing plants to PEG-induced moderate and severe drought conditions. Plant primary productivity was maintained under moderate drought conditions, due to the presence of alternative electron donors fueling the PSII. Additionally, the high anthocyanin production under drought conditions, act as photoprotective shields against photoinhibition. Moreover, the increased quinone turnover rate simultaneously with a net rate of RC closure and density increase, acted as a counteractive measure, allowing high energy fluxes into the photosystems under drought conditions. PSI showed an activity reduction, indicating that under drought conditions the ETC activity acts as an energetic escape route. Furthermore, membrane remodeling could also be observed under drought. The total fatty acid and omega-3 linolenic acid (18:3) contents were maintained, under osmotic stress. Membrane restructuring with lower amounts of polyunsaturated fatty acids (18:3) is considered an adaptation to osmotically stressful environments. Increased 18:1 and 16:1t fatty acids production improve the LHCs and chloroplast membrane stabilization, allowing the LHC to maintain its efficient functioning. The results here presented are very similar to the ones observed in the past regarding A. tripolium feedback to salinity stress, indicating that the mechanisms to overcome osmotic stress, either due to increased salinity or reduced water availability, are the same.

Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty acid composition.

A profound analysis of A. tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty acid profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty acid profile. Plants exposed to 125-250 mm NaCl showed higher photochemical light harvesting efficiencies and lower energy dissipation rates. under higher NaCl exposure, there is evident damage of the oxygen evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the energy fluxes entering the PS II, in spite of the high amounts of energy dissipated and the loss of PS II antennae connectivity. Energy dissipation was mainly achieved through the auroxanthin pathway. Total fatty acid content displayed a similar trend, being also higher under 125-250 mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic acid and palmitic acid allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250 mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of energy dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.

Ecophysiological constraints of Aster tripolium under extreme thermal events impacts: Merging biophysical, biochemical and genetic insights.

Cold and heat waves are phenomenon that occurs in higher frequency and intensity due to global climate changes. Commonly cultivated crop species are crucially affected by extreme weather events, and therefore alternative crops - such as halophytes - gain in agricultural interest. While halophytes are potentially able to cope with temperature extremes on the long term exposure, effects of temporary events such as cold and heat waves are not yet described. In order to unveil the effects of these altered thermal environments, Aster tripolium plants were subjected to cold (9/5 °C) and heat (42/38 °C) waves regimes during 3 days and its photochemical and biochemical traits evaluated. In the potential cash crop A. tripolium cold waves induced the gene expression of dehydrins in order to counteract desiccation and thus to prevent oxidative stress. Regulatory proteins on the RNA maturation level (Maturase K) were highly expressed. Heat stress induced the gene expression of the cystein protease gene; most likely to degrade misfolded proteins temporary. Both thermal treatments decreased the photosynthetic efficiency and capacity, driven by a loss in the connectivity between PSII antennae. Nevertheless the light absorption capacity was unaffected due to an increased RC closure net rate. Cold wave-treated individuals showed a decrease in the carotenoid pigmentation, except auroxanthin. In cold wave treated individuals the overall peroxidase activity was significantly increased. Data suggest that exposure to both, cold and heat wave treatment decreased the ecophysiological capacity of A. tripolium.

In vitro plant regeneration of Aster scaber via somatic embryogenesis.

We established an in vitro plant regeneration system via somatic embryogenesis of Aster scaber, an important source of various biologically active phytochemicals. We examined the callus induction and embryogenic capacities of three explants, including leaves, petioles, and roots, on 25 different media containing different combinations of α-naphthalene acetic acid (NAA) and 6-benzyladenine (BA). The optimum concentrations of NAA and BA for the production of embryogenic calli were 5.0 µM and 0.05 µM, respectively. Media containing higher concentrations of auxin and cytokinin (such as 25 µM NAA and 25 µM BA) were suitable for shoot regeneration, especially for leaf-derived calli, which are the most readily available calli and are highly competent. For root induction from regenerated shoots, supplemental auxin and/or cytokinin did not improve rooting, but instead caused unwanted callus induction or retarded growth of regenerated plants. Therefore, plant growth regulator-free medium was preferable for root induction. Normal plants were successfully obtained from calli under the optimized conditions described above. This is the first report of the complete process of in vitro plant regeneration of A. scaber via somatic embryogenesis.

Mycorrhizal symbiosis and local adaptation in Aster amellus: a field transplant experiment.

Many plant populations have adapted to local soil conditions. However, the role of arbuscular mycorrhizal fungi is often overlooked in this context. Only a few studies have used reciprocal transplant experiments to study the relationships between soil conditions, mycorrhizal colonisation and plant growth. Furthermore, most of the studies were conducted under controlled greenhouse conditions. However, long-term field experiments can provide more realistic insights into this issue. We conducted a five-year field reciprocal transplant experiment to study the relationships between soil conditions, arbuscular mycorrhizal fungi and plant growth in the obligate mycotrophic herb Aster amellus. We conducted this study in two regions in the Czech Republic that differ significantly in their soil nutrient content, namely Czech Karst (region K) and Ceske Stredohori (region S). Plants that originated from region S had significantly higher mycorrhizal colonisation than plants from region K, indicating that the percentage of mycorrhizal colonisation has a genetic basis. We found no evidence of local adaptation in Aster amellus. Instead, plants from region S outperformed the plants from region K in both target regions. Similarly, plants from region S showed more mycorrhizal colonisation in all cases, which was likely driven by the lower nutrient content in the soil from that region. Thus, plant aboveground biomass and mycorrhizal colonisation exhibited corresponding differences between the two target regions and regions of origin. Higher mycorrhizal colonisation in the plants from region with lower soil nutrient content (region S) in both target regions indicates that mycorrhizal colonisation is an adaptive trait. However, lower aboveground biomass in the plants with lower mycorrhizal colonisation suggests that the plants from region K are in fact maladapted by their low inherent mycorrhizal colonization. We conclude that including mycorrhizal symbiosis in local adaptation studies may increase our understanding of the mechanisms by which plants adapt to their environment.

Competition for patchy soil resources reduces community evenness.

The presence of small-scale patches of soil resources has been predicted to increase competition, because multiple species will proliferate roots in the same small area, and therefore decrease plant diversity. I tested whether such patches reduced species evenness in a community of four old-field species, both with and without interspecific interactions. In species mixtures, patches reduced evenness, while in "communities" constructed via combined monocultures, in which species did not compete, patches increased evenness. Therefore, the reduction in evenness in response to patches was due to changes in competition. Community-level changes may be attributable to plant foraging traits-in species with low foraging precision, competition reduced abundance much more in patchy soils than in even soils, while in species with high root foraging precision, the effect of competition was similar in patchy and even soils.

Consequences of near and far between-population crosses for offspring fitness in a rare herb.

Crosses between plants from different populations may result in heterosis or outbreeding depression. However, despite its importance for conservation, little is known about the spatial scale over which these effects may arise. To investigate the consequences of between-population crosses at two distinct spatial scales, we conducted reciprocal crosses between four populations from two regions in the rare perennial herb Aster amellus. We assessed seed set and offspring fitness in a common garden experiment. Overall, between-population crosses within regions (10 km) resulted in 8% lower seed set than within-population crosses, while between-region crosses (70 km) resulted in 17% higher seed set than within-population crosses. Moreover, offspring from between-population crosses produced 18% more flower heads than offspring from within-population crosses. We conclude that hybridisation between A. amellus plants from different populations did not lead to immediate outbreeding depression and, thus, could represent a valid conservation option to increase genetic diversity. Moreover, our results suggest that the distance between populations affects the outputs of between-population crosses and therefore needs to be taken into account when promoting gene flow between populations.

Root foraging traits and competitive ability in heterogeneous soils.

The responses of plant roots to nutrient patches in soil may be an important component of competitive ability. In particular, the scale, precision, and rate of foraging for patchy soil resources may influence competitive ability in heterogeneous soils. In a target-neighbor experiment in the field, per-individual and per-gram competitive effects were measured for six old-field species with known root foraging scale, precision, and rate. The presence and number of nutrient patches were also manipulated in a full factorial design. Number and presence of patches did not influence the outcome of competition. Competitive ability was not related to total plant size, growth rate, or root:shoot allocation, or to root foraging precision. Per-individual competitive effects were marginally correlated with root foraging scale (biomass of roots) and root foraging rate (time required to reach a patch). Therefore, competitive ability was more closely related to ability to quickly fill a soil volume with roots than to ability to preempt resource-rich patches.

Co-regulation of brassinosteroid biosynthesis-related genes during xylem cell differentiation.

To understand the regulatory mechanisms of brassinosteroid (BR) biosynthesis in specific plant developmental processes, we first investigated the accumulation profiles of BRs and sterols in xylem differentiation in a Zinnia culture. The amounts of many substances in the late C28 sterol biosynthetic pathway to campesterol (CR), such as episterol and 24-methylenecholesterol, as well as those in the BR-specific biosynthetic pathway from CR to brassinolide (BL), were elevated in close association with tracheary element differentiation. Among them, 6-deoxotyphasterol (6-deoxoTY) accumulated to unusually high levels within cells cultured in tracheary element-inductive medium, while castasterone (CS) was not elevated either within or outside cells. To identify the molecular basis of this co-up-regulation of BRs and C28 sterols, we isolated Zinnia genes for the key enzymes of BR biosynthesis, ZeSTE1, ZeDIM, ZeDWF4, ZeCPD1 and ZeCPD2. RNA gel blot analysis of these genes indicated a coordinated increase in transcripts for ZeSTE1, ZeDIM, ZeDWF4 and ZeCPD1, and a tracheary element differentiation-specific increase in transcripts for ZeDWF4 and ZeCPD1. In situ hybridization experiments of ZeDWF4 and ZeCPD1 mRNAs revealed their preferential accumulation in procambium cells, immature xylem cells and xylem parenchyma cells. These results suggest that BR biosynthesis during tracheary element differentiation may be regulated by the coordinated regulation of broad sterol biosynthesis and specific regulation of BR biosynthesis, which occurs in part by elevated transcript levels of genes encoding BR biosynthetic enzymes, specifically ZeDWF4 and ZeCPD1. These data provide new insights into the regulation of BR biosynthesis and BR signaling during plant development.

Distribution and ecology of cytotypes of the Aster amellus aggregates in the Czech Republic.

BACKGROUND AND AIMS: Polyploidy is viewed as an important mechanism of sympatric speciation, but only a few studies have documented patterns of distribution and ecology of different cytotypes in their contact zone. Aster amellus agg. (Asteraceae) is one of the species with documented multiple ploidy levels. The aim of this study was to determine spatial distribution and ecology of two cytotypes, diploid (2n = 18) and hexaploid (2n = 54), of Aster amellus agg. at their contact zone in the Czech Republic. METHODS: Root-tip squashes and flow cytometry were used to determine the ploidy of 2175 individuals from 87 populations. To test whether some differences in ecology between the two ploidy levels exist, in each locality relevés were recorded and abiotic conditions of the sites were studied by estimating potential direct solar radiation, Ellenberg indicator values and above-ground biomass. KEY RESULTS: Together with diploid and hexaploids, minorite cytotypes (triploid, pentaploid and nonaploid) were found. No significant ecological differences between diploid and hexaploid cytotypes were found. In spite of this, no population consisting of both of the two basic cytotypes was found. CONCLUSIONS: The results of this study show that the contact zone of diploid and hexaploid cytotypes in the Czech Republic is much more diffuse than indicated in previous records. Although populations of both cytotypes occur in close proximity (the closest populations of different cytotypes were 500 m apart), each individual population consists of only one basic ploidy level. This was unexpected since there are no clear differences in abiotic conditions between populations. Taken together with the absence of an intermediate tetraploid cytotype and with reference to published world distributional patterns of different ploidy levels, this suggests a secondary contact zone. Detailed genetic study is, however, necessary to confirm this.

Aster tripolium L. and Sesuvium portulacastrum L.: two halophytes, two strategies to survive in saline habitats.

Aster tripolium L. (Dollart, Germany) and Sesuvium portulacastrum L. (Dakhla, Morocco) are potential halophytic vegetables, fodder plants, and ornamentals for re-vegetating saline land. To compare their strategies involved in salt tolerance both plants were grown with 0%, 1.5%, and 3% (Aster) or 0%, 2.5%, and 5% (Sesuvium) NaCl in the watering solution. The growth rate was reduced in both species with increasing NaCl concentrations. The quotient of Na(+)/K(+) indicates that Aster accumulates more K(+) in comparison to Na(+) while the reverse is true for Sesuvium. Osmolality of the leaf sap increased with increasing NaCl concentration in both Aster and Sesuvium. Transpiration rate was severely reduced in both Aster (3%) and Sesuvium (5%) plants after 10 d of NaCl watering. The CO(2) assimilation rate decreased in Aster (3%) and Sesuvium (5%) NaCl-treated plants from day 5 to day 10. The most important results from chlorophyll fluorescence measurements were derived from the non-photochemical quenching analysis (NPQ). First, both plants had linearly increasing levels of NPQ with increasing NaCl concentrations. Second, Sesuvium had almost half the NPQ value when compared to Aster under increased soil salinity. In Aster P-ATPase activities were decreased in plants treated with 3% NaCl after three days of treatment, F-ATPase activities increased with increasing NaCl concentrations and no clear changes were measured in V-ATPase activities. In Sesuvium any changes could be observed in the three ATPase activities determined. To conclude, Aster and Sesuvium use different strategies in adaptation to soil salinity.

3-D physical models of mitosis (with asters) and cytokinesis.

First, we define new concepts of Life Objects, Informative Objects and Virtual Objects, Discrete Chromosome Rings (DCR); we introduce a mathematical concept of meridian plane (MP) in a three dimensional (3-D) cylindrical coordinate system (CCS). Based on these concepts, classic mechanics, classic electromagnetism and published biological data, we develop our 3-D physical models of natural and normal mitosis (with asters) and cytokinesis, for animal cells in M phase. We propose following hypotheses: Chromosomes Exclusion: No normally and naturally replicated chromosomes can occupy the same nucleus without growing sizes of the nucleus and the cell. Spontaneous and strong electromagnetic fields (EMF) forces among chromosomes, centrosomes and microtubules split the nucleus and separate the two sets of sister chromatids when they are strong enough. Nuclei Exclusion: No normally and naturally doubled nuclei can occupy the same cell if the doubled size of nuclei is not far smaller than size of the cell. The spontaneous and strong EMF forces in protoplasm (or cortex), separate two sets of chromosomes, spindles and poles, drive contractile proteins to the equator in cell cortex, and continue to guide and to transport free charged objects until complete the cytokinesis. Centrioles Exclusion: No naturally and normally doubled centrioles can occupy the same centrosome. The spontaneous and strong repulsive EMF forces are the primary cause for the exclusions. The principles of our models are also applied to mitosis and cytokinesis for lower plant cells, to that of multiple nuclei or mutant chromosomes, and to meiosis, for both animal cells and lower plant cells.

Photosynthetic limitations of a halophyte sea aster (Aster tripolium L) under water stress and NaCl stress.

To understand the mechanisms of salt tolerance in a halophyte, sea aster ( Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na(+) and Cl(-) were highest in NaCl-stressed leaves. Leaf osmotic potentials ( Psi(s)) were decreased by both stress treatments, whereas leaf turgor pressure ( Psi(t)) was maintained under NaCl stress. Decrease in Psi(s) without any loss of Psi(t) accounted for osmotic adjustment using Na(+) and Cl(-) accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO(2) fixation rate and O(2) evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Psi(t) and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na(+) and Cl(-), and that this plant has typical halophyte characteristics, but not drought tolerance.