Henquet M.G.L.,BU Bioscience |
Roelse M.,BU Bioscience |
Roelse M.,Laboratory of Plant Physiology |
de Vos R.C.H.,BU Bioscience |
And 8 more authors.
Metabolomics | Year: 2016
Introduction: Metabolomics has become a valuable tool in many research areas. However, generating metabolomics-based biochemical profiles without any related bioactivity is only of indirect value in understanding a biological process. Therefore, metabolomics research could greatly benefit from tools that directly determine the bioactivity of the detected compounds. Objective: We aimed to combine LC–MS metabolomics with a cell based receptor assay. This combination could increase the understanding of biological processes and may provide novel opportunities for functional metabolomics. Methods: We developed a flow through biosensor with human cells expressing both the TRPV1, a calcium ion channel which responds to capsaicin, and the fluorescent intracellular calcium ion reporter, YC3.6. We have analysed three contrasting Capsicum varieties. Two were selected with contrasting degrees of spiciness for characterization by HPLC coupled to high mass resolution MS. Subsequently, the biosensor was then used to link individual pepper compounds with TRPV1 activity. Results: Among the compounds in the crude pepper fruit extracts, we confirmed capsaicin and also identified both nordihydrocapsaicin and dihydrocapsaicin as true agonists of the TRPV1 receptor. Furthermore, the biosensor was able to detect receptor activity in extracts of both Capsicum fruits as well as a commercial product. Sensitivity of the biosensor to this commercial product was similar to the sensory threshold of a human sensory panel. Conclusion: Our results demonstrate that the TRPV1 biosensor is suitable for detecting bioactive metabolites. Novel opportunities may lie in the development of a continuous functional assay, where the biosensor is directly coupled to the LC–MS. © 2016, The Author(s).
van der Hooft J.J.J.,Wageningen University |
van der Hooft J.J.J.,Netherlands Metabolomics Center |
Vervoort J.,Wageningen University |
Vervoort J.,Netherlands Metabolomics Center |
And 5 more authors.
Metabolomics | Year: 2012
The identification of large series of metabolites detectable by mass spectrometry (MS) in crude extracts is a challenging task. In order to test and apply the so-called multistage mass spectrometry (MS n) spectral tree approach as tool in metabolite identification in complex sample extracts, we firstly performed liquid chromatography (LC) with online electrospray ionization (ESI)-MS n, using crude extracts from both tomato fruit and Arabidopsis leaf. Secondly, the extracts were automatically fractionated by a NanoMate LC-fraction collector/injection robot (Advion) and selected LC-fractions were subsequently analyzed using nanospray-direct infusion to generate offline in-depth MS n spectral trees at high mass resolution. Characterization and subsequent annotation of metabolites was achieved by detailed analysis of the MS n spectral trees, thereby focusing on two major plant secondary metabolite classes: phenolics and glucosinolates. Following this approach, we were able to discriminate all selected flavonoid glycosides, based on their unique MS n fragmentation patterns in either negative or positive ionization mode. As a proof of principle, we report here 127 annotated metabolites in the tomato and Arabidopsis extracts, including 21 novel metabolites. Our results indicate that online LC-MS n fragmentation in combination with databases of in-depth spectral trees generated offline can provide a fast and reliable characterization and annotation of metabolites present in complex crude extracts such as those from plants. © 2011 Springer Science+Business Media, LLC.
PubMed | BU Bioscience, Laboratory of Cell Biology, Laboratory of Plant Physiology and Netherlands Metabolomics Center
Type: | Journal: Metabolomics : Official journal of the Metabolomic Society | Year: 2016
Metabolomics has become a valuable tool in many research areas. However, generating metabolomics-based biochemical profiles without any related bioactivity is only of indirect value in understanding a biological process. Therefore, metabolomics research could greatly benefit from tools that directly determine the bioactivity of the detected compounds.We aimed to combine LC-MS metabolomics with a cell based receptor assay. This combination could increase the understanding of biological processes and may provide novel opportunities for functional metabolomics.We developed a flow through biosensor with human cells expressing both the TRPV1, a calcium ion channel which responds to capsaicin, and the fluorescent intracellular calcium ion reporter, YC3.6. We have analysed three contrasting Among the compounds in the crude pepper fruit extracts, we confirmed capsaicin and also identified both nordihydrocapsaicin and dihydrocapsaicin as true agonists of the TRPV1 receptor. Furthermore, the biosensor was able to detect receptor activity in extracts of both Our results demonstrate that the TRPV1 biosensor is suitable for detecting bioactive metabolites. Novel opportunities may lie in the development of a continuous functional assay, where the biosensor is directly coupled to the LC-MS.
Kohlen W.,Laboratory of Plant Physiology |
Charnikhova T.,Laboratory of Plant Physiology |
Bours R.,Laboratory of Plant Physiology |
Lopez-Raez J.A.,Laboratory of Plant Physiology |
Bouwmeester H.,Laboratory of Plant Physiology
Plant signaling & behavior | Year: 2013
Strigolactones are plant signaling molecules that induce germination of parasitic plant seeds, initiate host plant - arbuscular mycorrhizal fungus symbiosis and act as plant hormones controlling shoot branching and root architecture. To date four unique strigolactones (e.g., orobanchol, didehydroorobanchol isomers 1 and 2 and the aromatic strigolactone solanacol) have been reported in the root exudates and extracts of tomato (Solanum lycopersicum). Here we report on the presence of several additional strigolactones in tomato root exudates and extracts, orobanchyl acetate, two 7-hydroxyorobanchol isomers, 7-oxoorobanchol and two additional didehydroorobanchol isomers and discuss their possible biological relevance.
Menendez V.,Laboratory of Plant Physiology |
Abul Y.,Laboratory of Plant Physiology |
Bohanec B.,Center for Plant Biotechnology and Breeding |
Lafont F.,Mass Spectrometry Unit |
Fernandez H.,Laboratory of Plant Physiology
Acta Physiologiae Plantarum | Year: 2011
The fern Asplenium nidus L. is in great demand as an ornamental plant. The aim of this work was to investigate the influence of phytohormones in promoting a gametophytic and sporophytic growth in homogenized sporophytes tissue. Exogenous application of 0. 5 and 5 μM N6-benzyladenine, 0. 05 and 0. 5 μM indole-3-acetic acid (IAA), and 0. 3 and 3 μM gibberellic acid (GA3) favoured sporophyte regeneration, whereas gametophyte regeneration took place when plant material was cultured in a hormone-free liquid MS medium. The endogenous contents of the auxin IAA, the cytokinins trans-zeatin, trans-zeatin riboside, dihydrozeatin, dihydrozeatin riboside, isopentenyladenine and isopentenyladenosine, and the gibberellins GA1, GA3, GA4, GA7, GA9 and GA20 in growing gametophytes and sporophytes were evaluated. Similar levels of the auxin and cytokinins and qualitative differences in the gibberellins were found between both generations. © 2011 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.
Nikiforou C.,Laboratory of Plant Physiology |
Zeliou K.,Laboratory of Plant Physiology |
Kytridis V.-P.,Laboratory of Plant Physiology |
Kyzeridou A.,Laboratory of Plant Physiology |
Manetas Y.,Laboratory of Plant Physiology
Environmental and Experimental Botany | Year: 2010
Winter leaf reddening occurs in some plants as a response to the combination of cold temperatures and high light. Hence, a protective function against photoinhibition of photosynthesis has been suggested. However, it is unknown whether the leaf anthocyanic trait confers long term benefits to the plant and to that aim parameters related to fitness were measured. We took advantage of intra-species variation in the expression of the red leaf character displayed by the Mediterranean shrub Cistus creticus, a field of which during winter becomes a mosaic of green and red individuals under apparently similar environmental conditions. The red individuals are known to be more sensitive to winter photoinhibition of photosynthesis, hence anthocyanins might serve a compensatory function through light screening and/or detoxifying reactive oxygen species. If anthocyanins are indeed beneficial in this regard, this might be reflected in altered growth and reproductive output of red compared to green individuals. Both phenotypes displayed similar photosynthetic performance indices before and after the winter red period, yet reds suffered a considerable drop in this parameter concomitant with anthocyanin accumulation. This photosynthetic inferiority was irregularly linked to growth, since red plants produced fewer new leaves during the following spring, yet shoot relative growth rate was higher. Moreover both phenotypes displayed similar flower numbers, pollination success and seed yield, mass and germinability. As judged by the similar final reproductive output, vulnerability to the winter stress does not render the red phenotype less fit, nor anthocyanin accumulation render it more fit. Moreover, the photosynthetic inferiority of the red phenotype, although linked to slightly reduced leaf number, it was not limiting for reproductive effort and success. Regardless of function, winter leaf redness in C. creticus may indicate photosynthetically weak individuals. However, neither a fitness cost nor benefit of anthocyanins can inferred in this system. © 2009 Elsevier B.V. All rights reserved.
Continuous long-term uv-b irradiation reduces division and expansion of epidermal cells in true leaves, but accelerates developmental stages such as true leaf unfolding and male flower bud production in cucumber (Cucumis sativus L.) seedlings
Yamasaki S.,Laboratory of Plant Physiology |
Shigeto H.,Laboratory of Plant Physiology |
Ashihara Y.,Laboratory of Plant Physiology |
Noguchi N.,Laboratory of Plant Physiology
Environmental Control in Biology | Year: 2014
We examined the effects of 25 d of continuous ultraviolet-B (UV-B) irradiation (0.57 W m-2) on cucumber (Cucumis sativus L.) seedlings. The leaf areas of all unfolded true leaves and the plant height were reduced considerably by 25 d of UV-B irradiation. In each true leaf, both the epidermal cell area and the total epidermal cell number were reduced after UVB treatment. The expression of five cell cycle-related genes (Cs Cyclin A [CsCycA], CsCycB, CsCycD3;1, CsCycD3;2, and Cs Cyclin-dependent kinase A [CsCDKA]) in the shoot apical meristem (SAM) was reduced within 48 h of UV-B exposure. These results indicate that continuous UV-B irradiation inhibits the cell cycle in the SAM and thus reduces cell division and cell expansion in true leaves. On the other hand, the number of unfolded true leaves increased after exposure to UV-B for 15-25 d. Furthermore, 60% of the plants produced male flower buds after exposure to UV-B for 25 d. Therefore, continuous long-term UV-B irradiation affects the SAM of cucumber seedlings, resulting in reduced plant growth, but does accelerate various developmental stages.
Agency: Narcis | Branch: Project | Program: Completed | Phase: Agriculture | Award Amount: | Year: 2007
Agency: Narcis | Branch: Project | Program: Completed | Phase: Agriculture | Award Amount: | Year: 2007
News Article | April 21, 2016
Plant experts in the Netherlands can still become enthusiastic about a special variety of broomrape. Farmers in southern countries, however, are less impressed because broomrape and its 'sister' striga are considered a harmful weed in these regions. Research in the Laboratory of Plant Physiology of Wageningen University shows how useful insects that can control this weed may lend a helping hand in the future, namely via the plant's own aromatic substances. "You can also use a mixture of aromatic substances to distinguish various varieties of parasitic plants," says Harro Bouwmeester, professor in plant physiology. Several varieties of broomrape and striga are parasitic plants, requiring the root system of other plants to survive. As broomrape reaches its northern distribution limit in the Netherlands it is less common and causes fewer problems for farmers. In Southern Europe, the Middle East and Africa, however, the opposite is the case. There the parasitic plants can become a very harmful weed in crops such as tomatoes, sunflowers, maize and sorghum. In Israel tomato farmers are combating the broomrape with herbicides, for instance, but the plants themselves may offer a biological alternative. Each variety of broomrape emits a complex of over 100 types of volatile aromatic substances. Some of these aromatic signals are picked up by insects that are harmful to the plant as they lay their eggs in the ovaries, after which the larvae eat the young seeds. "If we can identify those specific aromatic substances, we would be able to lure the insects to the plants with extra aromas and deploy them as a potential biological control method," Harro Bouwmeester explains. While searching for useable aromatic substances, Peter Tóth, post doc in Bouwmeester's group, discovered another practical application of the aromatic substances from broomrape: they can help identify one variety from another. In taxonomy (the science involving the description and classification of plants) there is still uncertainty about the description and classification of the various broomrape varieties: which genus do they belong to? By looking at the composition of the aromatic mix, Tóth and his colleagues were able to classify a number of these 'disputed varieties' in the proper taxonomic genus. Their findings were described in the magazine Frontiers in Plant Science this month. "In addition, we found a difference between the aromas of parasitic and non-parasitic broomrape varieties," Bouwmeester continues. "This may teach us something about how these varieties evolved into weeds. Some varieties that have yet to become harmful have an aromatic composition resembling that of the harmful varieties. We believe this means that these varieties, too, will eventually become weeds." In time Bouwmeester hopes that the research into the aromatic substances of broomrape will lead to biological control of the weed. "For farmers in Southern Europe it could mean an alternative to the herbicides they use now. For farmers in Africa, who often cannot afford herbicides at all, this could be a sustainable alternative to labour-intensive weed control. They now try to remove the weeds by hand, with large parts of the harvests being lost as a result."