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

Natural resins and bioactive natural products thereof as potential anitimicrobial agents

Natural products and their derivatives have historically been invaluable as a source of therapeutic agents and have contributed to the discovery of antimicrobial agents. However, today with the development of drug-resistant strains, new scaffolds and new sources of bioactive compounds are needed. To this end, plant derived natural resins are reviewed for their potential application as antimicrobial agents. Natural gums, extracts of the whole resins, as well as specific extracts, fractions, essential oils and isolated compounds from the above resins are discussed in terms of their antifungal, antibacterial, and antiprotozoal activity. © 2011 Bentham Science Publishers Ltd

Synthesis of novel conformationally constrained pyrazolo[4,3-c]quinoline derivatives as potential ligands for the estrogen receptor

The preparation of three novel classes of pyrazolo[4,3-c]quinoline derivatives is reported. The easily accessible 2,3-dihydro-1H-quinolin-4-ones were used as the starting materials and were functionalized with three different acylating agents affording their respective constrained core substrates. The latter by condensation with phenyl(or 4-methoxyphenyl)hydrazines and subsequent debenzylation or demethylation provided the desired pyrazole derivatives. Some interesting features emerged with respect to the regioselectivity and mechanism of these reactions. © Georg Thieme Verlag Stuttgart

A two-season impact study on Globularia alypum: adaptive leaf structures and secondary metabolite variations

A detailed histo-anatomical and phytochemical study on Globularia alypum was carried out to investigate season-induced plant responses related to leaf morphology and content of secondary metabolites (SMs). Leaf tissue analysis by light and scanning electron microscopy showed neither morphological nor significant anatomical adaptions/variations to the coldest and the hottest seasons. Both summer and winter leaf tissue resulted amphistomatic with compact mesophyll and capitate glandular trichomes. Epidermal and mesophyll cells showed a significant accumulation of osmiophilic compounds, further characterized as phenolics by the histochemical screening. Beside the morpho-anatomical similarities in tissue arrangement, SMs production, within and among the different natural product classes, resulted seasonally dependent. In total, 24 SMs belonging to iridoids, flavonoids and phenylethanoid glycosides were tentatively assigned by UHPLC-HRMS analysis. Among iridoids, (E)-globularicisin showed to be the prevalent constituent, with content up to 70%. Total iridoid production was increased by 48.1% during summer. Similarly, flavonoid anabolism was more active during the same period, although with less remarkable seasonal variation reaching 25.6%. In contrast to the overall higher production of iridoids and flavonoids, a drastic fall (33.5%) of phenylethanoids was observed during summer. © 2018 Società Botanica Italiana

Leaf structure and histochemistry of Ficus carica (Moraceae), the fig tree

Heavily "armed" with trichomes, the hypostomatic leaf of Ficus carica L. is investigated with light and scanning electron microscopy. Histochemical tests were also applied to trace the secondary matabolites produced in the leaf of this well known, Mediterranean tree with the delicious fruits. Numerous idioblasts with crystals, laticifers and sized lythocysts appear within the compact mesophyll. Protective trichomes are located on both surfaces while capitate secretive hairs and stomata appear only abaxially. Stomata, concerning their magnitude, can be assembled in three size-groups in which the arrangement of their parastomatal cells differs as well. Histochemical reactions proved positive only for alkaloids within the laticifers and phenolic compounds, especially for the condensed tannins accumulated within the vacuole of the epidermal cells of this simply constructed leaf with unique features among the Mediterranean herbs. Comparison of GC-MS and LC-HRMS spectra of different extracts of the nerves and, separately, the rest of the leaf, revealed that coumarines such as umbeliferone, psoralen, bergapten and xanthotoxol are probably biosynthetically produced within the nerve area where the highly differentiated, main secretive apparatus of the leaf (laticifers) resides. That is why, these metabolites, are absent from the extracts of the undifferentiated cells derived from in vitro cultures. © 2015 Elsevier GmbH

Acretoside, a new sucrose ester from Aristolochia cretica

A new sucrose ester, acretoside, was isolated from the roots of the Greek endemic species Aristolochia cretica and identified as 6-O-p-coumaroyl-β-D-fructofuranosyl-(2 → 1)-α-D-glucopyranoside (1). In addition, a known sucrose ester, identified as arillatose B, two phenylpropanoid glucose esters, and five derivatives of aristolochic acid were isolated. Their structures were elucidated on the basis of MS and NMR data. © 2005 Taylor & Francis

Photo-activated DNA binding and antimicrobial activities of furoquinoline and pyranoquinolone alkaloids from Rutaceae

To find novel photo-active compounds of potential use in photochemotherapy from higher plants, photo-activated antimicrobial and DNA binding activities of the furoquinolines, skimmianine, kokusaginine, and haplopine, and a pyranoquinolone, flindersine, from two species of Rutaceae plants were investigated. TLC overlay assays against a methichillin-resistant strain of Staphylococcus aureus and Candida albicans were employed to test antimicrobial properties. All of the tested compounds showed photo-activated antimicrobial activity against S. aureus in the order of kokusaginine > haplopine, flindersine > skimmianine. Weaker activity was found for C. albicans. Photo-activated DNA binding activity of these compounds was investigated by a method using restriction enzymes and a specially designed 1.5 kb DNA fragment. Kokusaginine showed inhibition against all of the 16 restriction enzymes. Haplopine showed a similar inhibition pattern but the binding activity against Asc I and Sma I with restriction sequences consisting only of G and C was very weak. Skimmianine showed binding activity against Xba I, BciV I, Sal I, Pst I, Sph I and Hind III, but very weak or no activity was found for the other restriction enzymes. A pyranoquinolone, flindersine, showed no activity against any of the restriction enzymes. Photo-activated DNA binding activity of furoquinolines was therefore in the order of kokusaginine > haplopine > skimmianine, which was the same order as their photo-activated antimicrobial activity against S. aureus. Therefore, it was concluded that DNA is one of the cellular targets for the furoquinolines to exert their biological activities, similar to psoralens. However, because flindersine showed photo-activated antimicrobial activity against S. aureus but did not show photo-activated DNA binding activity, it is clear that there are other cellular target components for this compound to exert photo-toxic activity