Effects of Phytohormones and Jasmonic Acid on Glucosinolate Content in Hairy Root Cultures of Sinapis alba and Brassica rapa

Although some study have established hairy root cultures from brassicaceous plants with glucosinolates (GS) as characteristic secondary metabolite, studies are missing which compare hairy roots with the corresponding mother plants. Therefore, two different plant species—Sinapis alba and Brassica rapa subsp. rapa pygmeae teltoviensis—were transformed with the Agrobacterium rhizogenes strain A4. Aliphatic and indolyl GS were present in B. rapa, exhibiting larger quantities in leaves than in roots. Aromatic p-hydroxybenzyl GS were found particularly in the leaves of S. alba. However, the proportion of indolyl GS increased suddenly in transformed hairy roots of S. alba and B. rapa. Cultivation with the phytohormone kinetin (0.5 mg?L^?1) enhanced GS accumulation in B. rapa hairy roots, however not in S. alba, but 2,4-D (0.4 mg?L^?1) induced de-differentiation of roots in both species and reduced GS levels. GS levels especially of 1-methoxyindol-3ylmethyl GS increased in hairy roots in response to JA, but root growth was inhibited. While 2 weeks of cultivation in 100 to 200 μM JA were determined at optimum for maximum GS yield in S. alba hairy root cultures, 4 weeks of cultivation in 50 to 100 μM JA was the optimum for B. rapa.     

Evolution of Brassica rapa var. rapa L. volatile composition by HS-SPME and GC/IT-MS

Brassica rapa var. rapa L. (turnip) is highly appreciated and consumed by human. In this work, the volatile profile of B. rapa var. rapa was studied during the maturation process. The volatiles of seeds, sprouts with 6 and 9 days, and adult plant were determined using headspace solid-phase microextraction (HS-SPME) combined with gas chromatography/ion trap-mass spectrometry (GC/IT-MS).Several constituents, including alcohols, aldehydes, esters, ketones, norisoprenoids, nitrogen and sulphur compounds were characterized, totalizing 64 compounds. Isothiocyanates are the main volatiles in all matrices, being 3-butenyl isothiocyanate the major compound. Qualitative and quantitative differences were found among the analysed materials. Nitrogen and sulphur compounds decreased during the maturation process, while terpenes, aldehydes, norisoprenoids and ester compounds were present in higher amounts during germination, especially in sprouts with 9 days of development.     

Quercetin and Rutin as Modifiers of Aphid Probing Behavior

Rutin and its aglycone quercetin occur in the fruits, leaves, seeds, and grains of many plant species and are involved in plant herbivore interactions. We studied the effect of the exogenous application of rutin and quercetin on the probing behavior (= stylet penetration activities in plant tissues) of Acyrthosiphon pisum on Pisum sativum, Myzus persicae on Brassica rapa ssp. pekinensis, and Rhopalosiphum padi on Avena sativa using the electrical penetration graph technique (EPG = electropenetrography). The reaction of aphids to quercetin and rutin and the potency of the effect depended on aphid species, the flavonol, and flavonol concentration. Quercetin promoted probing activities of A. pisum within non-phloem and phloem tissues, which was demonstrated in the longer duration of probes and a trend toward longer duration of sap ingestion, respectively. M. persicae reached phloem in a shorter time on quercetin-treated B. rapa than on the control. Rutin caused a delay in reaching sieve elements by A. pisum and deterred probing activities of M. persicae within non-phloem tissues. Probing of R. padi was not affected by quercetin or rutin. The potency of behavioral effects increased as the applied concentrations of flavonols increased. The prospects of using quercetin and rutin in plant protection are discussed.     

Secoiridoids and other chemotaxonomically relevant compounds in Pedicularis: phytochemical analysis and comparison of Pedicularis rostratocapitata Crantz and Pedicularis verticillata L. from Dolomites

We compared the respective metabolite patterns of two Pedicularis species from Dolomites. Seven phenylethanoid glycosides, i.e., verbascoside (1), echinacoside (2), angoroside A (3), cistantubuloside B1 (4), wiedemannioside C (5), campneoside II (11) and cistantubuloside C1 (12), together with several iridoid glucosides as aucubin (6), euphroside (7), monomelittoside (8), mussaenosidic acid (9) and 8-epiloganic acid (13) were identified. Pedicularis verticillata showed also the presence of greatly unexpected secoiridoids, ligustroside (14) and excelside B (15), very rare compounds in Lamiales. Both PhGs and iridoids are considered of taxonomical relevance in the Asteridae and their occurrence in Pedicularis was discussed. In particular, the exclusive presence of several compounds such as 8-epiloganic acid (13), campneoside II (11), cistantubuloside C1 (12), ligustroside (14) and excelside B (15) in Pedicularis rostratocapitata, and angoroside A (3), cistantubuloside B1 (4) and wiedemannioside C (5) in P. verticillata could be considered specific markers for the two botanical entities.     

Unusual molecular pattern in Ajugoideae subfamily: the case of Ajuga genevensis L. from Dolomites

We analysed the ethanolic extract from Ajuga genevensis L. (Lamiaceae) growing in Dolomites, part of Italian Alps. Three new compounds for this species were identified: rosmarinic acid (1), oleanolic acid (2) and maslinic acid (3), representative of two different classes of chemical compounds (phenylpropanoids and pentacyclic triterpenes). A. genevensis resulted to be a valuable source of these compounds endowed with interesting biological activities (i.e. antioxidant, neuroprotective, anti-inflammatory, antiproliferative). The recognition of compounds (1), (2) and (3) may also confirm the ethnomedicinal uses of this plant. From a chemotaxonomical point of view, it is worth noting that iridoids were not evidenced in this accession. Iridoids are considered chemotaxonomic marker in Lamiales, and, in contrast with a previous study on this species, the presence of aucubin was not confirmed. In addition, the presence of large amounts of rosmarinic acid (1) was unexpected for a species that does not belong to subfamily Nepetoideae.     

Two new eremophilenolides from the roots of Ligulariopsis shichuana and their anti-phytopathogenic fungal and antifeedant activities

Two new eremophilenolides, ligushicins A (1) and B (2), and two known compounds including β-sitosterol and ursolic acid were isolated from Ligulariopsis shichuana. The structures of new compounds were established on the basis of 1D and 2D NMR data and HRESIMS data interpretation. The absolute configuration of new compounds was assigned by ECD spectroscopy, and that of ligushicins A (1) was confirmed by X-ray diffraction analysis. The antifungal and antifeedant activities of new compounds were evaluated against four plant pathogenic fungi and third-instar larvae of Plutella xylostella, respectively. Ligushicins A (1) and B (2) exhibited potent antifungal activity against Botrytis cinerea and Fusarium oxysporum with minimum inhibitory concentration (MIC) values ranging from 50 to 100 mg/L, while they also exhibited weak antifeedant activities.     

冻地银莲花中三萜皂苷的HPLC/MSn分析

药用植物冻地银莲花(Anemone rupestris ssp. gelida)的全草中含有丰富的三萜皂苷, 经HPLC-MSⁿ分析, 发现其总皂苷中共含有10余个三萜皂苷成分. 除由总离子流色谱图给出各皂苷成分的相对含量外, 从每个色谱峰的正负离子ESI-MS获得相应皂苷的分子量, 进而由多级质谱给出糖链连接的信息, 结合文献报道, 将11个主要三萜皂苷成分分别鉴定为革叶常春藤皂苷F (1), 牡丹草皂苷D (2), 刺楸皂苷B (3), 革叶常春藤皂苷E (4), 红毛七皂苷D (5), 常春藤皂苷B(6), 刺五加皂苷C₃ (7), 牡丹草皂苷B (8), 刺楸皂苷A (9), 木通皂苷D (10)和齐墩果酸-3-鼠李糖基-(12)-[葡萄糖基-(14)]-阿拉伯糖苷(11). 其中微量成分2, 5, 7, 10和11为首次从冻地银莲花中分离鉴定. 最后通过与标准品的HPLC保留时间对照证实了他们的存在.     

Identification of Iridoids,Flavonoids and Triterpenes from the Methanolic Extract of Melampyrum bihariense A. Kern. and the Antioxidant Activity of the Extract

Melampyrum bihariense A. Kern. (Scrophulariaceae), a plant species used in traditional medicine for the treatment of rheumatic disorders and skin infections, was investigated with regard to its antioxidant activity and identification of its bioactive chemical constituents. The crude methanolic extract of the aerial parts of M. bihariense was examined by the spectrophotometric DPPH (1,1-diphenyl-2-picrylhydrazyl) and ferric reducing antioxidant power methods. The free radical scavenging capacity (SC₅₀) of the extract was found by the DPPH method to be 27.10 mg mL⁻¹, and the ferric reducing ability equivalent to ascorbic acid at 50 mg mL⁻¹ was 0.709 μg mL⁻¹. The chemical composition of this highly effective in the methanolic extract was analysed, and the main compounds were isolated through solvent–solvent partition, and multiple chromatographic separations, including column chromatography, vacuum liquid chromatography, centrifugal planar chromatography and preparative thin-layer chromatography. The structures were established by one- and two-dimensional NMR and liquid chromatography-mass spectrometry. The iridoids aucubin (1), 8-epi-loganin (2) and mussaenoside (3), the flavones apigenin and luteolin and the triterpene acids ursolic acid and oleanolic acid were identified; components 2, 3, ursolic acid and oleanolic acid for the first time in this species. The present study reveals that M. bihariense exerts antioxidant activity, and the iridoids, flavonoids and triterpene acids may be the main bioactive constituents of its methanolic extract.

     

Alpha-glucosidase inhibitory activity of ethanol extract,fractions and purified compounds from the wood of Albizia myriophylla

Albizia myriophylla Benth. is a medicinal herb which is used as a traditional remedy for various ailments including diabetes in Thailand. In our continued investigation of the biological activity of A. myriophylla, the ethanol extract, fractions and the isolated compounds from the wood of this plant were evaluated for in vitro α-glucosidase inhibition using spectrophotometric method. The plant ethanol extract and its different fractions possessed α-glucosidase inhibitory activity in a concentration-dependent manner. Dichloromethane fraction of the wood ethanol extract exhibited the highest percent inhibition against α-glucosidase (69.30%) among all fractions. Subsequent α-glucosidase inhibition assay proved that indenoic acid (1), 8-methoxy-7, 3′,4′-trihydroxyflavone (2) and 3,4,7,3′-tetrahydroxyflavan (3) were partially rational for antidiabetic effect of this plant species. Among these compounds, 3 (IC₅₀ 98.59 μg/mL) exhibited potent inhibition of α-glucosidase, compared with a positive control acarbose (IC₅₀ 125 μg/mL). The inhibitory effect towards α-glucosidase of compounds 1–3 was reported herein for the first time.     

The use of fast protein liquid chromatography with ICP-OES and ES-MS-MS detection for the determination of various forms of aluminium in the roots of Chinese cabbage

The distribution of aluminium (Al) species was investigated in the roots of Al-tolerant Chinese cabbage (Brassica rapa L. ssp. pekinensis) by employing fast protein liquid chromatography (FPLC) with inductively coupled plasma optical emission spectrometry (ICP-OES) and electrospray tandem mass spectrometry (ES-MS-MS) detection. The cabbage was exposed to a nutrient solution that contained 10 μg cm⁻³ of Al³⁺. The results demonstrated that after 24 h of exposure, Al was quantitatively taken up by the cabbage and was distributed in different parts of the plant. 36 ± 6% of total Al was located in the roots, while the remaining 64 ± 10% was transferred to the leaves. It was found that in the roots Al was partially present in the root sap (15.5%), while the majority (84.5%) was accumulated in its apoplasmic compartments. It was further demonstrated that the proportion of Al that entered the symplasm formed a complex with organic acid. Speciation analysis by FPLC with ICP-OES detection and ES-MS-MS identification of the binding ligand indicated that Al-citrate complex was the prevailing species in the root sap.The results of the present study showed that both immobilization of Al in the apoplasmic compartments of the roots and transformation of Al³⁺ to Al-citrate are most likely responsible for the tolerance of Chinese cabbage (B. rapa L. ssp. pekinensis) to the toxic effects of Al³⁺.     

Polyphenolic contents and antioxidant activities of aerial parts of Salvia multicaulis from the Iran flora

Abstract

Folk medicine sometimes involves the use of salvia species for therapeutic purposes. Polyphenols with the highest amounts included rosmarinic acid (7.358mg/g), catechin (1.5?mg/g), vanillin (1.00?mg/g), chlorogenic acid (0.53?mg/g), quercetin (0.16?mg/g), and p-coumaric acid (0.015). Furthermore, the results showed that S. multicaulis has a high content of total phenol (4.39?mg/g) and DPPH activity (8.44?mg/g). Salvia multicaulis could be potentially used as a medicinal plant because of its antioxidant activity and polyphenol content.     

Phenolic glycosides of Juncus acutus and its anti-eczematic activity

The total alcohol extract of Juncus acutus L. showed significant anti-eczematic activity. The isolation of the five phenolic glycosides which responsible for this activity were isolated and identified as oxyresveratrol 2-O-β-D-glucopyranoside (1), resveratrol 3′,4′-O,O′-di-β-D-glucopyranoside (2), markhamioside F (3), canthoside B (4), and caffeic acid glucorhamnoside (5). The toxic effect of the alcohol extract of the plant was studied on mice to determine their LD₅₀, which proved to be nontoxic up to 3000 mg/kg body weight. The anti-eczematic activity of the isolated compounds was tested in mice and showed variable effect. Compounds 3 and 4 were found to have the highest activity; they cured eczema by 90 and 100% respectively. Published in Khimiya Prirodnykh Soedinenii, No. 2, pp. 125–127, March–April, 2006.     

Chemical composition and antioxidant activity of a polar extract of Thymelaea microphylla Coss. et Dur.

Thymelaea microphylla Coss. et Dur. (Thymelaeaceae) is a rare medicinal plant endemic to Algeria. In order to continue our studies on this species, herein we report the isolation and characterisation of 20 compounds from a hydroalcoholic extract (EtOH–H₂O 7:3) of the aerial parts. They include monoterpene glucosides (1–3), phenolic acid derivatives (4, 8 and 9), phenylpropanoid glucosides (5 and 6), flavonoids (7, 10 and 11), a benzyl alcohol glucoside (12), ionol glucosides (13–16), lignans (17–19) and a bis-coumarin (20). All the structures were elucidated by spectroscopic methods including 1D and 2D NMR experiments, as well as ESI-MS analysis. Moreover, the extract of T. microphylla showed a significant and concentration-dependent free radical-scavenging activity in vitro, correlated to the presence of phenolic and chlorogenic acid derivatives (8, 9 and 4).     

Two new diacetylene glycosides: bhutkesoside A and B from the roots of Ligusticopsis wallichiana

Two new diacetylene glycosides: bhutkesoside A (1) and B (2), along with 10 known compounds, i.e. falcarindiol (3), chlorogenic acid (4), 5-O-p-coumaroyl-quinic acid (5), 3,5-di-O-caffeoyl-quinic acid (6), 4-hydroxy-7-methoxy-phenylethanol (7), ferulic acid (8), dehydrodiconiferyl alcohol-4-O-β-d-glucopyranoside (9), 5,7-dihydroxy-2-methylchromone-7-O-rutinoside (10), schumanniofioside B (11) and marmesinin (12) were isolated from the roots of Ligusticopsis wallichiana (DC) Pimenov & Kljuykov (Apiaceae), commonly known as ‘Bhutkesh’ in Nepal. The structures were determined on the basis of spectroscopic data. Compounds 4 and 6 showed potent antioxidant activity on DPPH free radical scavenging assay.     

Nepetadiol,a new triterpenediol from <Emphasis Type="Italic">Nepeta suavis</Emphasis>

Nepetadiol, a new tetracyclic triterpene diol (1), was isolated from the chloroform-soluble portion of the whole plant of Nepeta suavis. In addition, lawsonin (2) and lawsonic acid (3) have been isolated for the first time from this species. The structures of the isolated compounds were based on ¹H and ¹³C NMR spectra, including two-dimensional NMR techniques like COSY, HMQC, and HMBC, and comparison with the literature data.     

Triterpenoids and lignans from the stems of Schisandra glaucescens

Two new triterpenoids, 24(E)-3α,12α-dihydroxyl-lanost-9(11),24-dien-26-oid acid (1) and 3,4-seco-24(Z)-6α,7β-dihydroxyl-cycloart-4(28),24-dien-3,26-dioic-3-methyl ester (2), and ten known ones (3–12), together with eight lignans (13–20), were isolated from the stems of Schisandra glaucescens. Their structures were established on the basis of extensive spectroscopic analyses. Compounds 1–3, 5–7, 9, 11–13 and 15–18 were isolated from this species for the first time. 1 and 2 were evaluated for their in vitro cytotoxicities against the mouse cancer B16 cell line, human cancer HepG2 and MCF7 cell lines by MTT assay.     

Isolation and antiproliferative activity of triterpenoids and fatty acids from the leaves and stem of Turraea vogelii Hook. f. ex benth

Chloroform extract from the leaves of Turraea vogelii Hook f. ex Benth demonstrated cytotoxic activity against a chronic myelogenous leukemia cell, K-562 with IC₅₀ of 14.27 μg/mL, while chloroform, ethyl acetate and methanol extracts from the stem of the plant inhibited K-562 cells growth with IC₅₀ of 19.50, 24.10 and 85.40 μg/mL respectively. Bioactive chloroform extract of Turraea vogelii leaves affords two triterpenoids: oleana-12,15,20-trien-3β-ol (1), and oleana-11,13-dien-3β,16α,28-triol (2), with six fatty esters, ethyl hexaeicos-5-enoate (3), 3-hydroxy-1,2,3-propanetriyltris(tetadecanoate) (4), 1,2,3-propanetriyl(7Z,7′Z,7′′Z)tris(-7-hexadecenoate) (5), 1,2,3-propanetriyl(5Z,5′Z,5′′Z)tris(-5-hexadecenoate) (6), 1,2,3-propanetriyltris(octadecanoate) (7), and 2β-hydroxymethyl tetraeicosanoate (8). Tetradecane (9), four fatty acids: hexadecanoic acid (10), tetradecanoic acid (11), (Z)-9-eicosenoic acid (12), and ethyl tetradec-7-enoate (13) were isolated from chloroform extract of Turraea vogelii stem. 1,2,3-propanetriyltris(heptadecanoate) (14), (Z)-9-octadecenoic acid (15) and (Z)-7-tetradecenoic acid (16) were isolated from ethyl acetate extract while (Z)-5-pentadecenoic acid (17) was obtained from methanol extract of the plant stem. Compounds 1, 2, 5, 6, 11, 12, 15, 16 and 17 exhibited pronounced antiproliferative activity against K-562 cell lines.     

GC-MS and GC-FID analyses of the volatile constituents of Brachychiton rupestris and Brachychiton discolor,their biological activities and their differentiation using multivariate data analysis

Abstract

The volatile constituents of the leaves and flowers of Brachychiton rupestris and Brachychiton discolor (Malvaceae) were characterized by GC-FID and GC-MS. A total of 92 compounds were identified. The major component of B. rupestris leaves was α-farnesene (34.57%) while in the flowers it was n-heptacosane (29.5%). (8Z, 11Z, 14Z)-heptadecatrienal (21.2%) in the leaves and palmitic acid (83.49%) in the flowers represent the abundant ingredients of B. discolor. Unsupervised pattern recognition techniques of multivariate data analysis exemplified by hierarchical cluster analysis (HCA) and principle component analysis (PCA) relying upon chromatographic results effectively discriminate the two related species. Also, both investigated species were assessed for their cytotoxic and antimicrobial efficacy. Results revealed that B. discolor flowers showed the highest cytotoxicity against human lung carcinoma (A-549) cell lines with IC₅₀ equals 7.98?μg/mL, whereas B. discolor leaves exhibited the highest antimicrobial potency against all tested microorganisms namely Streptococcus pneumonia, Bacillus subtilis, Escherichia coli as well as Aspergillus fumigatus and Geotrichum candidum (MICs between 0.98 and 15.63?µg/mL) except for Salmonella typhi.     

Chemical analysis of the semipurified extract of Paullinia cupana and evaluation of in vitro inhibitory effects against Helicobacter pylori

Abstract

Paullinia cupana Kunth, commonly known as guarana, is a native Brazilian plant species from the Amazon area that presents various biological effects, including antimicrobial action. The aim of this study was to chemically analyse the semipurified aqueous extract (AqF) of the plant and to evaluate the activity of crude (CE), ethyl-acetate (EAF), and AqF extracts against Helicobacter pylori. The chemical profile of AqF was determined based on solid analysis ¹³C-NMR, direct infusion mass spectrometry (ESI-MS), and MALDI-TOF. The ¹³C-NMR spectrum showed characteristics of flavan-3-ol and oligomeric proanthocyanidins. ESI-MS revealed the presence of procyanidin, caffeic acid and its derivatives. MALDI-TOF analysis detected procyanidins of up to 6 units and profisetinidins of up to 5 units. Whereas CE and EAF showed inhibitory activity against H. pylori, CE, EAF, and AqF presented not high inhibitory activity against urease. The results demonstrate the potential of P. cupana to control and prevent H. pylori infection.