Micropropagation and molecular characterization of Hypericum perforatum L. subsp. veronense (Schrank) H. Lindb., a valuable medicinal plant with ornamental value

St. John's wort (Hypericum perforatum) is a perennial species with a worldwide interest due to its pharmaceutical characteristics which is also of ornamental value for gardening with native elements. Given that in vitro culture allows mass multiplication of selected genotypes, we investigated the micropropagation of the ex situ cultivated Greek native H. perforatum subsp. veronense using novel substances and its genetic fingerprinting. Apart from common auxins and cytokinins, a culture medium (BB) was used for the first time for its micropropagation and the effect of betaine, PVP, SNP and triacontanol were explored for the first time. Betaine addition of 200 mg/L produced proliferated shoots and roots without the need of plant growth regulators although a 0.5 mg L-1 BA was the best treatment in proliferation experiments. Shoot number was highest in 0.5 BA + 0.1 IBA (mg L-1), in 1 μΜ sodium nitroprusside (SNP) + 1 mg L-1 IAA+200 mg L-1 betaine and in PVP (0.5-1.5 mg L-1) + 1 mg L-1 IBA + 200 mg L-1. Greater SNP concentration had either no differences or negative effect, while the greatest root number were recorded in Control. All three treatments of triacontanol (1-10 μg L-1) + 0.5 mg L-1 BA + 200 mg L-1 betaine resulted in higher shoot number. However, the statistically greatest root number were recorded in low concentration of SNP, or in absence of PVP and triacontanol. The micro-plants of H. perforatum subsp. veronense obtained from the SunCapTM sealed vessels and transplanted into the soil substrates presented 90.2% success; they were then transplanted to 1 L pots and were successfully established in the field for cultivation. The molecular characterization of H. perforatum subsp. veronense was achieved with DNA barcoding fingerprints deposited in the GenBank regarding four molecular markers of cpDNA (petB/petD, rbcL, trnL/trnF and rpoC1)

Endophytic Bacteria From the Roots of the Medicinal Plant Alkanna tinctoria Tausch (Boraginaceae): Exploration of Plant Growth Promoting Properties and Potential Role in the Production of Plant Secondary Metabolites

Alkannin and shikonin (A/S) are enantiomeric naphthoquinones produced in the roots of certain plants from the Boraginaceae family such as Lithospermum spp. and Alkanna spp. They possess antimicrobial, anti-tumoral and wound healing properties. The production of secondary metabolites by Alkanna tinctoria might be influenced by its endomicrobiome. To study the interaction between this medicinal plant and its bacterial endophytes, we isolated bacteria from the roots of wild growing Alkanna tinctoria collected near to Athens and Thessaloniki in Greece. Representative strains selected by MALDI-TOF mass spectrometry were identified by partial 16S rRNA gene sequence analysis. In total, 197 distinct phylotypes of endophytic bacteria were detected. The most abundant genera recovered were Pseudomonas, Xanthomonas, Variovorax, Bacillus, Inquilinus, Pantoea, and Stenotrophomonas. Several bacteria were then tested in vitro for their plant growth promoting activity and the production of cell-wall degrading enzymes. Strains of Pseudomonas, Pantoea, Bacillus and Inquilinus showed positive plant growth properties whereas those of Bacteroidetes and Rhizobiaceae showed pectinase and cellulase activity in vitro. In addition, bacterial responses to alkannin and shikonin were investigated through resistance assays. Gram negative bacteria were found to be resistant to the antimicrobial properties of A/S, whereas the Gram positives were sensitive. A selection of bacteria was then tested for the ability to induce A/S production in hairy roots culture of A. tinctoria. Four strains belonging to Chitinophaga sp., Allorhizobium sp., Duganella sp., and Micromonospora sp., resulted in significantly more A/S in the hairy roots than the uninoculated control. As these bacteria can produce cell-wall degrading enzymes, we hypothesize that the A/S induction may be related with the plant-bacteria interaction during colonization

The electronic trade in endemic plants of Cyprus through the Internet

<p>This study assesses the current situation regarding the electronic trade of the Cypriot endemic plants. We examine the plant material sold over the Internet, prices and vendors, and we explore to what extent this activity complies with national laws and international treaties. Overall, 48 taxa (34% of the Cypriot endemic flora) are traded by 21 nurseries of Europe and North America, belonging mostly to Lamiaceae, Brassicaceae, Iridaceae, Asparagaceae, Fabaceae and Asteraceae. Seeds are the commonest form of sale; prices range from €0.12 to 0.79 per seed and from €0.31 to 2.56 per gram. The price range for live plants is €5.67–30.22 (highest for <i>Quercus alnifolia</i>). Among the traded taxa, two are designated critically endangered, another two endangered, four vulnerable, 12 are globally rare and threatened and 12 are protected at the international level. None of the nurseries surveyed seem to have been granted a permit by the authorities of Cyprus to harvest and sell its wild flora, at least during the last decade. State authorities should extend the current legislative framework so as to cover all endemic and nationally threatened taxa, monitor their wild populations, engage in routine surveillance of their e-trade and regulate access to the country’s genetic resources.</p

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. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Non-polar secondary metabolites and essential oil of ex situ propagated and cultivated Sideritis syriaca L. subsp. syriaca (Lamiaceae) with consolidated identity (DNA Barcoding): towards a potential new industrial crop

Over the past decade, perennial plants of the genus Sideritis L. have attracted great scientific interest and they have become the subject of several studies, including the European Union&apos;s herbal monograph. The present study aimed to explore the identity of a biotype of S. syriaca subsp. syriaca documented with DNA barcoding, investigated its macro- and micro- propagation and to study the chemical profile of the dichloromethane extract, as well as of the Essential Oil (EO) of the ex situ propagated and cultivated plant material. An improved in vitro propagation protocol, as well as pioneer effective propagation with cuttings (both at commercially acceptable rates) are provided for a DNA-barcoded biotype of Sideritis syriaca L. subsp. syriaca (malotira, Cretan mountain tea) which is a single-island endemic plant of Crete (Greece), with declining wild populations due to over collection. From the aerial parts of ex situ propagated and cultivated material (harvested in July) two phytosterols (β-sitosterol and stigmasterol), two ent-kaurene diterpenes; siderol and eubotriol, as well as one methylated flavone (xanthomicrol) have been isolated. This is the first time that eubotriol has been isolated from this taxon. The structures of the isolated compounds have been established by spectroscopic means and the complete Nuclear Magnetic Resonance (NMR) assignments for eubotriol are presented. Moreover, the essential oil was obtained by hydrodistillation of the plant material during the flowering stage and was analyzed by Gas Chromatography-Mass Spectrometry (GC–MS). Forty-four compounds were identified. The monoterpene hydrocarbons constituted the major fraction of the essential oil (34.2%), while β-phellandrene (18.5%) and kaur-15-ene (or ent-kaur-15-ene; 17.3%) were the main compounds. In conclusion, the above results document the first necessary steps taken for the ex situ conservation and sustainable exploitation of a new industrial crop with promising potential. © 2020 Elsevier B.V

An experimental brackish aquaponic system using juvenile gilthead sea bream (Sparus aurata) and rock samphire (Crithmum maritimum)

Brackish aquaponics using Mediterranean fish and plants provides an alternative opportunity for a combined production of high-quality food products with high commercial and nutritional value. This is the first study that investigates the effect of two different salinities (8 and 20 ppt) on growth and survival of Sparus aurata and Crithmum maritimum along with the cellular stress pathways using the activation of heat shock proteins (HSPs) and mitogen-activated protein kinase (MAPK) protein family members and the water bacterial abundance. In total, 156 fish were used (average initial weight of 2.55 g, length of 5.57 cm) and 36 plants (average initial height of 8.23 cm) in floating racks above the 135 L fish tanks. Survival rate for both organisms was 100%. C. crithmum grew better at 8 ppt (t-test, p &lt; 0.05). The growth rate of S. aurata was similar for both treatments (p &gt; 0.05). HSPs and MAPK were differentially expressed, showing tissue-specific responses. The average bacterial abundance at the end of the experiment was higher (p &lt; 0.05) in the 20 ppt (18.6 ± 0.91 cells × 105/mL) compared to the 8 ppt (6.8 ± 1.9 cells × 105/mL). The results suggest that the combined culture of euryhaline fish and halophytes provides good quality products in brackish aquaponics systems. © 2019 by the authors

Endophytic Bacteria From the Roots of the Medicinal Plant Alkanna tinctoria Tausch (Boraginaceae): Exploration of Plant Growth Promoting Properties and Potential Role in the Production of Plant Secondary Metabolites

Alkannin and shikonin (A/S) are enantiomeric naphthoquinones produced in the roots of certain plants from the Boraginaceae family such as Lithospermum spp. and Alkanna spp. They possess antimicrobial, anti-tumoral and wound healing properties. The production of secondary metabolites by Alkanna tinctoria might be influenced by its endomicrobiome. To study the interaction between this medicinal plant and its bacterial endophytes, we isolated bacteria from the roots of wild growing Alkanna tinctoria collected near to Athens and Thessaloniki in Greece. Representative strains selected by MALDI-TOF mass spectrometry were identified by partial 16S rRNA gene sequence analysis. In total, 197 distinct phylotypes of endophytic bacteria were detected. The most abundant genera recovered were Pseudomonas, Xanthomonas, Variovorax, Bacillus, Inquilinus, Pantoea, and Stenotrophomonas. Several bacteria were then tested in vitro for their plant growth promoting activity and the production of cell-wall degrading enzymes. Strains of Pseudomonas, Pantoea, Bacillus and Inquilinus showed positive plant growth properties whereas those of Bacteroidetes and Rhizobiaceae showed pectinase and cellulase activity in vitro. In addition, bacterial responses to alkannin and shikonin were investigated through resistance assays. Gram negative bacteria were found to be resistant to the antimicrobial properties of A/S, whereas the Gram positives were sensitive. A selection of bacteria was then tested for the ability to induce A/S production in hairy roots culture of A. tinctoria. Four strains belonging to Chitinophaga sp., Allorhizobium sp., Duganella sp., and Micromonospora sp., resulted in significantly more A/S in the hairy roots than the uninoculated control. As these bacteria can produce cell-wall degrading enzymes, we hypothesize that the A/S induction may be related with the plant-bacteria interaction during colonization. © Copyright © 2021 Rat, Naranjo, Krigas, Grigoriadou, Maloupa, Alonso, Schneider, Papageorgiou, Assimopoulou, Tsafantakis, Fokialakis and Willems