DNA Barcoding and Fertilization Strategies in Sideritis syriaca subsp. syriaca, a Local Endemic Plant of Crete with High Medicinal Value.

Herein, we applied DNA barcoding for the genetic characterization of Sideritis syriaca subsp. syriaca (Lamiaceae; threatened local Cretan endemic plant) using seven molecular markers of cpDNA. Five fertilization schemes were evaluated comparatively in a pilot cultivation in Crete. Conventional inorganic fertilizers (ChFs), integrated nutrient management (INM) fertilizers, and two biostimulants were utilized (foliar and soil application). Plant growth, leaf chlorophyll fluorescence, and color were assessed and leaf content of chlorophyll, key antioxidants (carotenoids, flavonoids, phenols), and nutrients were evaluated. Fertilization schemes induced distinct differences in leaf shape, altering quality characteristics. INM-foliar and ChF-soil application promoted yield, without affecting tissue water content or biomass partitioning to inflorescences. ChF-foliar application was the most stimulatory treatment when the primary target was enhanced antioxidant contents while INM-biostimulant was the least effective one. However, when the primary target is yield, INM, especially by foliar application, and ChF, by soil application, ought to be employed. New DNA sequence datasets for the plastid regions of petB/petD, rpoC1, psbK-psbI, and atpF/atpH were deposited in the GenBank for S. syriaca subsp. syriaca while the molecular markers rbcL, trnL/trnF, and psbA/trnH were compared to those of another 15 Sideritis species retrieved from the GenBank, constructing a phylogenetic tree to show their genetic relatedness.

Adaptability of Wild-Growing Tulips of Greece: Uncovering Relationships between Soil Properties, Rhizosphere Fungal Morphotypes and Nutrient Content Profiles.

Wild-growing Greek tulips are protected plants but almost nothing is known about their natural nutrient status and rhizosphere fungal morphotypes in the wild, thus no insight is currently available into their growth and adaptation to their natural environment or artificial settings. To this end, several botanical expeditions were conducted with a special collection permit, and 34 tulip and soil samples were collected, representing 13 species from two phytogeographical regions of Greece (North Aegean Islands, Crete Island) and seven regions of mainland Greece. The tulips' content in essential macro- and micro-nutrients, respective physicochemical soil properties, and rhizosphere fungal morphotypes were assessed across samples, and all parameters were subjected to appropriate statistical analysis to determine their interrelationships. The results showed that soil variables played a significant role in shaping tulips' nutrient content, explaining up to 67% of the detected variability as in the case of phosphorus (P) in the above-ground plant tissue. In addition, significant correlations were observed (with an r value of up to 0.65, p < 0.001) between essential nutrients in the tulips, such as calcium (Ca) and boron (B). The principal component analysis (PCA) revealed that between the three spatial units examined, the total variability of tulips' nutrient content produced a clear distinction among sampled species, while the first two PCA axes managed to explain 44.3% of it. This was further confirmed by the analysis of variance (ANOVA) results which showed corresponding significant differences (at p < 0.05) in both the tulips' nutrient content and the studied soil properties as well (mean values of N, P, and K in the North Aegean Islands tulips' nutrient content, up to 53%, 119%, and 54% higher compared to those of the Crete Island, respectively). Our study sheds light on Greek tulips' adaptability and resilience in their original habitats, facilitating at the same time the undertaken efforts regarding their conservation and potential domestication in artificial settings.

Decoding the potential of a new Pseudomonas putida strain for inducing drought tolerance of tomato (Solanum lycopersicum) plants through seed biopriming.

A total of 11 potential plant growth promoting rhizobacteria previously isolated from naturally stressed environments were evaluated for various traits of interest for a beneficial symbiosis with plants, including colonization ability, biofilm formation, motility, exopolysaccharide production and salt tolerance. The vast majority of the strains were found to possess multiple plant growth promoting traits. Nevertheless, the intensity varied among isolates, with those originated from tomato plants being more efficient colonizers. The strain SAESo11, genetically characterized as a Pseudomonas putida member was selected for further investigation of its potential to alleviate drought stress in tomato seedlings. Inoculation with SAESo11 mitigated the negative effects of drought stress as indicated by growth and photosynthetic indices. Furthermore, bacterial inoculation enhanced H2O2 content and malondialdehyde levels in colonized plants. Drought treatment did not further alter the oxidative status of these plants. Similarly, total phenolic content and antioxidant enzyme activity were induced in plant tissues in response to drought stress only at the absence of inoculum. These results indicated that inoculation with the selected strain imposed plants at a priming state, that enabled them to respond more robustly at the exposure to drought stress and efficiently attenuated the drought-induced injury. This state of plant alertness mediated by SAESo11 occurred at no cost to growth, highlighting its role as a potential plant priming agent.

Greek Tulips: Worldwide Electronic Trade over the Internet, Global Ex Situ Conservation and Current Sustainable Exploitation Challenges.

From an ornamental viewpoint, tulips are famous clonally propagated crops. This research focuses on 15 wild-growing Greek tulip species including 11 range-restricted species, i.e., six Greek endemics and five Balkan or Aegean endemics and subendemics, among which seven are currently threatened with extinction (two Critically Endangered, three Endangered and two Vulnerable). An overview of the global electronic trade over the internet is presented herein for these valuable phytogenetic resources in an attempt to define the extent of their commercialization (25 nurseries in three countries, mainly bulb trade at various prices) with concomitant conservation implications. In the frame of the repatriation initiatives launched, their global ex situ conservation is overviewed according to the PlantSearch facility of the Botanic Gardens Conservation International (materials from 15 species stored in 41 botanic gardens of 14 countries). The results of this study on the Greek tulips showed that there are both well-established value chains and gaps in the market regarding the "botanical tulips"; revealed the compromised effectiveness of ex situ conservation for the majority of them; raised conservation concerns related to authorized access to these wild phytogenetic resources; and indicated that their future utilization should comply with the provision of national and international legislation. All these are envisaged and discussed within the framework of the newly launched research project TULIPS.GR which aims to be the pilot establishment of a national collection regarding all Greek tulips (currently holding 38 accessions of 13 species, including almost all of the threatened ones). The project's scope is to enable the creation of a sustainable value chain for the Greek tulips with authorized collections, sustainable conservation schemes, production of DNA barcoded propagation material, species-specific propagation and cultivation protocols, mycorrhizal investigations, field studies, applying innovative precise soil/foliar fertigation, and investigation of the postharvest treatment of fresh cut flowers, promoting networking and synergies with producers and associations in Greece and abroad.

Bacterial Communities in the Rhizosphere and Phyllosphere of Halophytes and Drought-Tolerant Plants in Mediterranean Ecosystems.

The aim of the study was to investigate the bacterial community diversity and structure by means of 16S rRNA gene high-throughput amplicon sequencing, in the rhizosphere and phyllosphere of halophytes and drought-tolerant plants in Mediterranean ecosystems with different soil properties. The locations of the sampled plants included alkaline, saline-sodic soils, acidic soils, and the volcanic soils of Santorini Island, differing in soil fertility. Our results showed high bacterial richness overall with Proteobacteria and Actinobacteria dominating in terms of OTUs number and indicated that variable bacterial communities differed depending on the plant's compartment (rhizosphere and phyllosphere), the soil properties and location of sampling. Furthermore, a shared pool of generalist bacterial taxa was detected independently of sampling location, plant species, or plant compartment. We conclude that the rhizosphere and phyllosphere of native plants in stressed Mediterranean ecosystems consist of common bacterial assemblages contributing to the survival of the plant, while at the same time the discrete soil properties and environmental pressures of each habitat drive the development of a complementary bacterial community with a distinct structure for each plant and location. We suggest that this trade-off between generalist and specialist bacterial community is tailored to benefit the symbiosis with the plant.

Plant growth promoting rhizobacteria isolated from halophytes and drought-tolerant plants: genomic characterisation and exploration of phyto-beneficial traits.

Plant growth promoting rhizobacteria (PGPR) are able to provide cross-protection against multiple stress factors and facilitate growth of their plant symbionts in many ways. The aim of this study was to isolate and characterize rhizobacterial strains under natural conditions, associated with naturally occurring representatives of wild plant species and a local tomato cultivar, growing in differently stressed Mediterranean ecosystems. A total of 85 morphologically different rhizospheric strains were isolated; twenty-five exhibited multiple in vitro PGP-associated traits, including phosphate solubilization, indole-3-acetic acid production, and 1-aminocyclopropane-1-carboxylate deaminase activity. Whole genome analysis was applied to eight selected strains for their PGP potential and assigned seven strains to Gammaproteobacteria, and one to Bacteroidetes. The genomes harboured numerous genes involved in plant growth promotion and stress regulation. They also support the notion that the presence of gene clusters with potential PGP functions is affirmative but not necessary for a strain to promote plant growth under abiotic stress conditions. The selected strains were further tested for their ability to stimulate growth under stress. This initial screening led to the identification of some strains as potential PGPR for increasing crop production in a sustainable manner.

Use of clay minerals for sewage sludge stabilization and a preliminary assessment of the treated sludge's fertilization capacity.

Preserving sewage sludge's N is important for its agronomic use and this could possibly be achieved by treating sludge with certain clay minerals. Nine clay minerals and additionally Ca(OH)2 were added to dewatered sewage sludge at 0-30 % rates (wet weight basis) (treatments). After 70 days of equilibration, all mixtures were analyzed for certain properties and the mineral-sludge mixtures which showed the highest microbial load reduction were further assayed, along with the limed and untreated sludge. From all minerals' treatments, the fecal indicators of sludge treated with 30% of two bentonites, attapulgite, saponite-attapulgite, and zeolite decreased considerably compared to the control. These treatments were performed also well regarding sludge's retention capacity of available inorganic N, with the attapulgite and zeolite treatments containing the significantly highest amounts of NO3-N and NH4-N, respectively. For the water-soluble inorganic N, similar results were obtained for the zeolite treatment, whereas the treatments with the two bentonites had the significantly highest NO3-N content. Also, considerable amounts of water-soluble P were obtained in all cases of the treated sludge with minerals. Limed sludge had the lowest content of the water-soluble inorganic N and P. As far as the micronutrients are concerned, only Zn and B were detectable in the water-soluble fraction of all five minerals' treatments. The heavy metals, which regulate sludge's agronomic use, were far below the respective permissible limits and lower than the untreated sludge, except for Ni and Cr in the attapulgite and saponite-attapulgite treatments. In conclusion, certain clay minerals, i.e., bentonite, attapulgite, mixed clay of saponite and attapulgite, and zeolite, seem promising materials for the stabilization of sewage sludge in the perspective of using them as a fertilizer. In addition, they seem to have higher fertilizing value than limed sludge. However, environmental (in respect of Ni and Cr) and agricultural (in respect of Zn and B) impacts must be considered.

Soil application of sewage sludge stabilized with steelmaking slag and its effect on soil properties and wheat growth.

The effect of sewage sludge, stabilized with steelmaking slag, on soil chemical properties and fertility and on wheat (Triticum aestivum L.) growth was evaluated. Dewatered sewage sludge [75% (wet weight basis)] stabilized with steelmaking slag (25%) and three soils with different pH values were used in a pot experiment with winter wheat. The following treatments were applied: (i) sludge addition of 30gkg-1 (≈ 120Mgha-1, rate equivalent to the common inorganic N fertilization for wheat, based on sludge's water soluble NO3-N), (ii) sludge addition of 10gkg-1 (≈ 40Mgha-1, rate equivalent to the common inorganic N fertilization for wheat, based on sludge's Kjeldahl-N), (iii) addition of the common inorganic N fertilization for wheat (120kgNha-1) as NH4NO3, (iv) control (no fertilizer, no sludge). Sludge application at both rates to all soils resulted in a significant increase of pH, electrical conductivity of the saturation extract (ECse) and soil available NO3-N and P, in comparison to the other two treatments and this increase remained constant till the end of the pot experiment. In sludge treatments pH did not exceed the critical value of 8.5, whereas ECse, although it did not reach the limit of 4dSm-1, exceeded the value of 2dSm-1 at the rate of 30gkg-1. Concentrations of heavy metals, which regulate the agronomic use of sewage sludge according to the established legislation, ranged from not detectable to lower than the respective permissible levels. Both rates of sludge's addition in all soils improved wheat's growth, as judged by the significant increase of the aboveground biomass yield and the total plant uptake of almost all nutrients, compared to the other two treatments. It was concluded that sewage sludge stabilized with steelmaking slag could be used in agriculture, applied at rates based on sludge's Kjeldahl-N content and crop's demand for N. However, potential environmental impacts must also be considered.

Metabolomic and physico-chemical approach unravel dynamic regulation of calcium in sweet cherry fruit physiology.

Calcium (Ca2) nutrition has a significant role in fruit physiology; however, the underlying mechanism is still unclear. In this study, fruit quality in response to CaCl2, applied via foliar sprays (Ca2) or/and hydro-cooling water (CaHC), was characterized in 'Lapins' cherries at harvest, just after cold storage (20 days at 0 °C) as well as after cold storage followed by 2 days at 20 °C, herein defined as shelf-life period. Data indicated that pre- and post-harvest Ca2+ applications increased total Ca2+ and cell wall bound Ca2+, respectively. Treatment with Ca reduced cracking whereas Ca + CaHC condition depressed stem browning. Both skin penetration and stem removal were affected by Ca2+ feeding. Also, several color- and antioxidant-related parameters were induced by Ca2+ treatments. Metabolomic analysis revealed significant alterations in primary metabolites among the Ca2+ treatments, including sugars (eg., glucose, fructose), soluble alcohols (eg., arabitol, sorbitol), organic acids (eg.,malate, quinate) and amino acids (eg., glycine, beta-alanine). This work helps to improve our knowledge on the fruit's response to Ca2+ nutrition.

Deregulation of apoplastic polyamine oxidase affects development and salt response of tobacco plants.

Polyamine (PA) homeostasis is associated with plant development, growth and responses to biotic/abiotic stresses. Apoplastic PA oxidase (PAO) catalyzes the oxidation of PAs contributing to cellular homeostasis of reactive oxygen species (ROS) and PAs. In tobacco, PAs decrease with plant age, while apoplastic PAO activity increases. Our previous results with young transgenic tobacco plants with enhanced/reduced apoplastic PAO activity (S-ZmPAO/AS-ZmPAO, respectively) established the importance of apoplastic PAO in controlling tolerance to short-term salt stress. However, it remains unclear if the apoplastic PAO pathway is important for salt tolerance at later stages of plant development. In this work, we examined whether apoplastic PAO controls also plant development and tolerance of adult plants during long-term salt stress. The AS-ZmPAO plants contained higher Ca2+ during salt stress, showing also reduced chlorophyll content index (CCI), leaf area and biomass but taller phenotype compared to the wild-type plants during salt. On the contrary, the S-ZmPAO had more leaves with slightly greater size compared to the AS-ZmPAO and higher antioxidant genes/enzyme activities. Accumulation of proline in the roots was evident at prolonged stress and correlated negatively with PAO deregulation as did the transcripts of genes mediating ethylene biosynthesis. In contrast to the strong effect of apoplastic PAO to salt tolerance in young plants described previously, the effect it exerts at later stages of development is rather moderate. However, the different phenotypes observed in plants deregulating PAO reinforce the view that apoplastic PAO exerts multifaceted roles on plant growth and stress responses. Our data suggest that deregulation of the apoplastic PAO can be further examined as a potential approach to breed plants with enhanced/reduced tolerance to abiotic stress with minimal associated trade-offs.

Silencing S-Adenosyl-L-Methionine Decarboxylase (SAMDC) in Nicotiana tabacum Points at a Polyamine-Dependent Trade-Off between Growth and Tolerance Responses.

Polyamines (PAs) are nitrogenous molecules that are indispensable for cell viability and with an agreed-on role in the modulation of stress responses. Tobacco plants with downregulated SAMDC (AS-SAMDC) exhibit reduced PAs synthesis but normal levels of PA catabolism. We used AS-SAMDC to increase our understanding on the role of PAs in stress responses. Surprisingly, at control conditions AS-SAMDC plants showed increased biomass and altered developmental characteristics, such as increased height and leaf number. On the contrary, during salt stress AS-SAMDC plants showed reduced vigor when compared to the WT. During salt stress, the AS-SAMDC plants although showing compensatory readjustments of the antioxidant machinery and of photosynthetic apparatus, they failed to sustain their vigor. AS-SAMDC sensitivity was accompanied by inability to effectively control H2O2 levels and concentrations of monovalent and divalent cations. In accordance with these findings, we suggest that PAs may regulate the trade-off between growth and tolerance responses.

Use of magnesia for boron removal from irrigation water.

The risk of B phytotoxicity due to high levels of B in irrigation water can be avoided by removing B from the water, before its use, through adsorption on certain adsorbents, such as magnesia (industrial MgO), if the latter can be proven to be an effective and easy to handle means for B removal. In addition, if such a material is applied as a fertilizer after its use and the adsorbed B is easily released into the soil solution, B phytotoxicity could constitute a potential hazard. The objectives of this work were to: (a) establish the optimum working conditions (equilibration time, solution to adsorbent ratio, and particle size of the adsorbent) for B adsorption, (b) assess the magnitude of B adsorption by magnesia, both in capacity and intensity terms, as well as the influence of temperature, (c) study B desorbability from magnesia, spiked with B at two rates, 5 and 0.5 mg g(-1), and (d) compare the results from b and c to those obtained using reagent grade MgO. The results showed that the time to achieve equilibrium depended on the B concentration of the external solution and ranged from 6 h (for B /= 50 mg L(-1)). The percentage of B adsorbed decreased as the volume of external solution to adsorbent increased and a working ratio of 50:1 was selected. For magnesia, B adsorption was particle size dependent with the smallest fraction (<0.1 mm) sorbing more B than the other three fractions studied (0.1-1.0, 1.1-2.0, 2.1-4.0 mm). Boron adsorption was conducted under strongly alkaline pH (10.3 +/- 0.2 and 10.4 +/- 0.1 for the reagent and magnesia, respectively) and increased with temperature. Both adsorbents exhibited a high B adsorption capacity (Langmuir maximum values were 5.85 +/- 0.39 and 4.45 +/- 1.31 mg B g(-1) for the reagent and magnesia, respectively) comparable to other metal oxides. However, the reagent grade MgO seemed to be superior to magnesia in terms of capacity and strength of B retention. This superiority of the reagent was attributed to its greater surface area (34.7 compared with 5.8 m(2) g(-1) for magnesia) and to its conversion to Mg(OH)(2) during the adsorption process, whereas magnesia remained unaltered, as was evident from X-ray diffractograms. Based on this data, magnesia seems to be an effective means for removing excess B from irrigation water, particularly if a material of fine particle size is used. Boron desorbability after 240 h of desorption time was more pronounced for magnesia reaching up to 55 and 60% of the amount of B added, at the spiked rates of 5 and 0.5 mg g(-1), respectively. Although these figures indicate that approximately half of the amount of B added remained adsorbed, they cannot be easily extrapolated to field conditions, and if B-laden magnesia is applied to soils, the possibility of B phytotoxicity cannot be excluded.