Metabolic responses of Thellungiella halophila/salsuginea to biotic and abiotic stresses: metabolite profiles and quantitative analyses.

The metabolite profiles of the model crucifer Thellungiella salsuginea (salt cress) ecotype Shandong subjected to various biotic and abiotic stresses were analyzed using HPLC-DAD-ESI-MS. Two different cruciferous microbial pathogens, Albugo candida, a biotrophic oomycete, and Leptosphaeria maculans, a necrotrophic fungus, elicited formation of the phytoalexins wasalexins A and B without causing visual damage on inoculated leaves. Analyses of non-polar and polar metabolites led to elucidation of the chemical structures of five metabolites: 4'-O-(E)-sinapoyl-7-methoxyisovitexin-2''-O-beta-D-glucopyranoside, 4'-O-(E)-sinapoylisovitexin-2''-O-beta-D-glucopyranoside, 4-O-beta-D-glucopyranosyl-7-hydroxymatairesinol, 5'-O-beta-D-glucopyranosyldihydroneoascorbigen and 3-O-beta-D-glucopyranosylthiane. 3-O-beta-D-glucopyranosylthiane, an unique metabolite for which we suggest the name glucosalsuginin, is proposed to derive from the glucosinolate glucoberteroin. In addition, the identification of a broad range of polar metabolites identical to those of other crucifers was carried out. Quantification of several metabolites over a period of eight days showed that concentrations of the polar phytoanticipin 4-methoxyglucobrassicin increased substantially in leaves irradiated with UV light (lambda(max) 254 nm) relative to control leaves, but not in leaves subjected to other stresses.

Photochemical dimerization of wasalexins in UV-irradiated Thellungiellahalophila and in vitro generates unique cruciferous phytoalexins.

The production of phytoalexins in Thellungiella halophila exposed to UV-radiation and NaCl was investigated over a period of 8 days. UV-radiation induced substantially larger quantities of wasalexins A and B than NaCl irrigation or CuCl(2) spray. Isolation of two metabolites and their chemical structure determination using X-ray diffraction analysis provided the phytoalexins biswasalexins A1 and A2, that resulted from head-to-tail photodimerization of wasalexin A. The production of biswasalexins A1 and A2 in stressed T. halophila, as well as their chemical synthesis and antifungal activity are reported. Biswasalexins A1 and A2 (60 nmol/g and 15 nmol/g fresh wt, respectively, 2 days after UV elicitation) are cruciferous phytoalexins whose formation in planta appears to result from a photochemical reaction, which might protect the plant from fungal attack and UV-radiation.

Metabolic changes in roots of the oilseed canola infected with the biotroph Plasmodiophora brassicae: phytoalexins and phytoanticipins.

Analyses of metabolite production and accumulation in roots of canola ( Brassica napus L. spp. oleifera) infected with the phytopathogen Plasmodiophora brassicae (clubroot) allowed the identification of 45 metabolites. HPLC analysis corroborated by metabolite isolation and NMR spectroscopic data demonstrated for the first time that phytoalexins and phytoanticipins were produced in roots of canola infected with a soilborne biotroph. In addition, six new indolyl metabolites were identified, synthesized, and tested against three fungal pathogens of canola. Multivariate data analysis using principal component analysis (PCA) revealed distinct metabolic responses of canola to P. brassicae infection during a six-week period. At late harvest days (five and six weeks), a clear clustering was observed among samples of infected roots because of the higher concentration of phytoalexins, while higher concentration of phytoanticipins contributed to the differentiation between three and four weeks samples of infected and control roots. Altogether, the data shows that canola roots under biotrophic attack are able to produce a complex blend of phytoalexins and other antimicrobial metabolites as a defensive response and that the metabolic regulation of phytoanticipins and phytoalexins appeared to correlate with the infection period.

Phytoalexins and polar metabolites from the oilseeds canola and rapeseed: differential metabolic responses to the biotroph Albugo candida and to abiotic stress.

The metabolites produced in leaves of the oilseeds canola and rapeseed (Brassica rapa L.) inoculated with either different races of the biotroph Albugo candida or sprayed with CuCl(2) were determined. This investigation established consistent phytoalexin (spirobrassinin, cyclobrassinin, and rutalexin) and phytoanticipin (indolyl-3-acetonitrile, arvelexin, caulilexin C, and 4-methoxyglucobrassicin) production in canola and rapeseed in response to both biotic and abiotic elicitation. In addition, a wide number of polar metabolites were isolated from infected leaves, including six new phenylpropanoids and two new flavonoids. The extractable chemical components of zoosporangia of A. candida and the anti-oomycete activity of phytoalexins were determined as well. Overall, the results suggest that during the initial stage of the interaction, leaves of B. rapa have a similar response to virulent and avirulent races of A. candida, with respect to the accumulation of chemical defenses. After this stage, despite the higher phytoalexin concentration, the "compatible" races could overcome the plant defense system for further infection, but growth of the "incompatible" races was inhibited. Since results of bioassays showed that cyclobrassinin and brassilexin were more inhibitory to A. candida than rutalexin, the apparent redirection of the phytoalexin pathway towards rutalexin, avoiding cyclobrassinin and brassilexin accumulation might be caused by the pathogen. Alternatively, A. candida might be able to detoxify both cyclobrassinin and brassilexin, similar to necrotrophic plant pathogens. Overall, the correlation between phytoalexin production in infected or stressed leaves and the outcome of the plant-pathogen interaction suggested that A. candida was able to elude the plant defense mechanisms by, for example, redirecting the phytoalexin biosynthetic pathway.

Efficient synthesis of brussalexin A, a remarkable phytoalexin from Brussels sprouts.

The synthesis of brussalexin A, the first phytoalexin containing an allyl thiolcarbamate group, and its selective inhibitory activity against fungal plant pathogens is described.

The first naturally occurring aromatic isothiocyanates, rapalexins A and B, are cruciferous phytoalexins.

The discovery of the first naturally occurring aromatic isothiocyanates, indole-3-isothiocyanates, their first synthesis, antimicrobial activity and proposed biogenetic origin in canola plants are reported.

Detection, characterization and identification of crucifer phytoalexins using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry.

We have analyzed 23 crucifer phytoalexins (e.g. brassinin, dioxibrassinin, cyclobrassinin, brassicanals A and C) by HPLC with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS) using both negative and positive ion modes. Positive ion mode ESI-MS appeared more sensitive than negative ion mode ESI-MS in detecting this group of compounds. A new HPLC separation method, new LC-MS and LC-MS(2) data and proposed fragmentation pathways, LC retention times, and UV spectra for selected compounds are reported.

Dracaenogenins A and B, new spirostanols from the red resin of Dracaena cochinchinensis.

A 12(13-->14)abeospirostanol dracaenogenin A (1) and a spirostanol dracaenogenin B (2) were isolated from Chinese dragon's blood, the red resin of Dracaena cochinchinensis (Agavaceae). Their structures were established as (14S,25R)12(13-->14)abeospirosta-5,13(18)-diene-1beta,3beta,15alpha-triol (1) and (25R) spirost-5-ene-1beta,3beta,14alpha,15alpha-tetrol (2) by means of spectroscopic analysis, especially by 2D NMR spectra, and X-ray crystallographic analysis. Dracaenogenin A (1) is the first example of a 12(13-->14)abeospirostane spirostanoid found in nature. Its biogenesis from ruscogenin (3) through namogenin (4) and 2 was tentatively proposed.

Steroidal saponins from fresh stem of Dracaena cochinchinensis.

A further phytochemical investigation on the fresh stem of Dracaena cochinchinensis yielded 18 steroidal saponins. Fourteen of which are new compounds, designated as 25(R,S)-dracaenosides E-H, M, O-Q, dracaenosides I-L, R, and 25(S)-dracaenoside N. Their structures were determined by detailed spectroscopic analysis, including extensive 1D and 2D NMR data, and the result of hydrolytic reaction.

Pregnane glycosides from Dracaena cochinchinensis.

Two new pregnane glycosides, named dracaenoside C and D, were isolated from the fresh stems of Dracaena cochinchinensis. Their structures were established as 3beta,14alpha-dihydroxypregna-5,16(17)-diene-20-one 3-O-alpha-L-rhamnopyranosyl(1 --> 2)[alpha-L-rhamnopyranosyl(1 --> 4)]-beta-D-glucopyranoside (1) and 3beta,14alpha-dihydroxypregna-5,16(17)-diene-20-one 3-O-alpha-L-rhamnopyranosyl(1 --> 2)[beta-D-glucopyranosyl(1 --> 3)]-beta-D-glucopyranoside (2) by means of 2D NMR spectral and chemical methods.