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.

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.