Dirk Selmar

Stress Enhances the Synthesis of Secondary Plant Products: The Impact of Stress-Related Over-Reduction on the Accumulation of Natural Products

Spice and medicinal plants grown under water deficiency conditions reveal much higher concentrations of relevant natural products compared with identical plants of the same species cultivated with an ample water supply. For the first time, experimental data related to this well-known phenomenon have been collected and a putative mechanistic concept considering general plant physiological and biochemical aspects is presented. Water shortage induces drought stress-related metabolic responses and, due to stomatal closure, the uptake of CO2 decreases significantly. As a result, the consumption of reduction equivalents (NADPH + H(+)) for CO2 fixation via the Calvin cycle declines considerably, generating a large oxidative stress and an oversupply of reduction equivalents. As a consequence, metabolic processes are shifted towards biosynthetic activities that consume reduction equivalents. Accordingly, the synthesis of reduced compounds, such as isoprenoids, phenols or alkaloids, is enhanced.

Compartmentation of cyanogenic glucosides and their degrading enzymes

Whereas high activities of β-glucosidase occur in homogenates of leaves of Hevea brasiliensis Muell.-Arg., this enzyme, which is capable of splitting the cyanogenic monoglucoside linamarin (linamarase), is not present in intact protoplasts prepared from the corresponding leaves. Thus, in leaves of H. brasiliensis the entire linamarase is located in the apoplasmic space. By analyzing the vacuoles obtained from leaf protoplasts isolated from mesophyll and epidermal layers of H. brasiliensis leaves, it was shown that the cyanogenic glucoside linamarin is localized exclusively in the central vacuole. Analyses of apoplasmic fluids from leaves of six other cyanogenic species showed that significant linamarase activity is present in the apoplasm of all plants tested. In contrast, no activity of any diglucosidase capable of hydrolyzing the cyanogenic diglucoside linustatin (linustatinase) could be detected in these apoplasmic fluids. As described earlier, any translocation of cyanogenic glucosides involves the interaction of monoglucosidic and diglucosidic cyanogens with the corresponding glycosidases (Selmar, 1993a, Planta 191, 191–199). Based on this, the data on the compartmentation of cyanogenic glucosides and their degrading enzymes in Hevea are discussed with respect to the complex metabolism and the transport of cyanogenic glucosides.

Evaluation of leaf water status by means of permittivity at terahertz frequencies

We present an electromagnetic model of plant leaves which describes their permittivity at terahertz frequencies. The complex permittivity is investigated as a function of the water content of the leaf. Our measurements on coffee leaves (Coffea arabica L.) demonstrate that the dielectric material parameters can be employed to determine the leaf water status and, therefore, to monitor drought stress in plant leaves. The electromagnetic model consists of an effective medium theory, which is implemented by a third order extension of the Landau, Lifshitz, Looyenga model. The influence of scattering becomes important at higher frequencies and is modeled by a Rayleigh roughness factor.

Transport of cyanogenic glucosides : linustatin uptake by Hevea cotyledons

The 14C-labelled cyanogenic glucosides linustatin (diglucoside of acetone cyanohydrin) and linamarin (monoglucoside of acetone cyanohydrin), prepared by feeding [14C]valine to plants of Linum usitatissimum L., were applied to cotyledons of Hevea brasiliensis Muell.-Arg. in order to study their transport. Both [14C]-linustatin and [14C]linamarin were efficiently taken up by the cotyledons. Whereas 14C was recovered completely when [14C]linustatin was applied to the seedling, only about one-half of the radioactivity fed as [14C]linamarin could be accounted for after incubation. This observation is in agreement with the finding that apoplasmic linamarase hydrolyzes linamarin but not the related diglucoside linustatin. These data prove that, in vivo, linamarin does not occur apoplasmically and that linustatin, which is exuded from the endosperm, is taken up by the cotyledons very efficiently. Thus, these findings confirm the linustatin pathway (Selmar et al. 1988, Plant Physiol. 86, 711–716), which describes mobilization and transport of the cyanogenic glucoside linamarin, initiated by the glucosylation of linamarin to yield linustatin. When linustatin is metabolized to non-cyanogenic compounds, in Hevea this cyanogenic diglucoside is hydrolyzed by a diglucosidase which splits off both glucose molecules simultaneously as one gentiobiose moiety (Selmar et al. 1988). In contrast, [14C]linustatin, which is taken up by the cotyledon, is not metabolized but is reconverted in high amounts to the monoglucosidic [14C]linamarin, which then is temporarily stored in the cotyledons. These data demonstrate that in Hevea, besides the simultaneous diglucosidase, there must be present a further diglucosidase which is able to hydrolyze cyanogenic diglucosides sequentially by splitting off only the terminal glucose moiety from linustatin to yield linamarin. From this, it is deduced that the metabolic fate of linustatin, which is transported into the source tissues, depends on the activities of the different diglucosidases. Whereas sequential cleavage — producing linamarin — is purely a part of the process of linamarin translocation (using linustatin as the transport vehicle), simultaneous cleavage, producing acetone cyanohydrin, is part of the process of linamarin metabolization in which the nitrogen from cyanogenic glucosides is used to synthesize non-cyanogenic compounds.

Stress metabolism in green coffee beans (Coffea arabica L.): expression of dehydrins and accumulation of GABA during drying.

In order to produce tradeable standard green coffee, processed beans must be dried. The drying procedure affects the abundance of relevant aroma substances, e.g. carbohydrates. Using molecular tools, the corresponding metabolic basis is analyzed. A decrease in water potential of the still living coffee seeds induces massive drought stress responses. As a marker for these stress reactions, accumulation of a general stress metabolite, GABA (gamma-aminobutyric acid), and associated gene expression of drought stress-associated dehydrins were monitored. The results of this study indicate that metabolism in drying coffee beans is quite complex since several events trigger accumulation of GABA. The first peak of GABA accumulation during drying is correlated with expression of isocitrate lyase and thus with ongoing germination processes in coffee seeds. Two subsequent peaks of GABA accumulation correspond to maxima of dehydrin gene expression and are thought to be induced directly by drought stress in the embryo and endosperm tissue, respectively. Apart from the significance for understanding basic seed physiology, metabolic changes in coffee seeds during processing provide valuable information for understanding the role and effect of the steps of green coffee processing on the quality of the resulting coffee.

Occurrence of lotaustralin in the genus Hevea and changes of HCN-potential in developing organs of Hevea brasiliensis

Abstract The mean HCN-potential (HCN-p) of freshly collected seeds of Hevea brasiliensis is 104.8 μmol HCN per g dry weight. More than 90% of the cyanogenic compound is stored in the endosperm. During seedling development under aseptic conditions HCN-p of the entire seedling decreases to 15% within 19 days. The cyanogenic compounds are metabolized during germination to form noncyanogenic substances. Leaves of H. pauciflora, H. benthanaana, H. pauciflora x H. guianensis and H. spruceana contain both linamarin and (R)-lotaustralin, whereas lotaustralin was not detectable in leaves and seeds of H. brasiliensis.

Selenium and nano-selenium in plant nutrition

Abstract Selenium (Se) is a naturally occurring metalloid element which occurs nearly in all environments. Se is considered as a finite and non-renewable resource on the Earth. The common sources of Se in earth’s crust occur in association with sulfide minerals such as metal selenide, whereas it is rarely found in elemental form (Se0). While there is no evidence of Se need for higher plants, several reports show that when Se added at low concentrations, Se exerts beneficial effects on plant growth. Se may act as quasi-essential micronutrient through altering different physiological and biochemical traits. Thus, plants vary considerably in their physiological and biochemical response to Se. This review focusses on the physiological importance of Se forms as well as different Se fertilizers for higher plants, especially plant growth, uptake, transport, and metabolism.

Dhurrin-6'-glucoside, a cyanogenic diglucoside from Sorghum bicolor.

A novel cyanogenic diglucoside has been isolated from methanolic extracts of young seedlings of Sorghum bicolor. Its structure was established as dhurrin-6-glucoside from NMR, mass spectrometry and enzymatic hydrolysis data. Compared with dhurrin, which is the major cyanogenic glucoside in sorghum leaves, dhurrin-6-glucoside occurs only in low concentrations. In contrast, however, the diglucoside is present in significant amounts in guttation droplets of young Sorghum seedlings. The presence of the diglucoside and its occurrence in apoplasmic exudates supports the hypothesis that diglucosides represent metabolites of cyanogenic monoglucosides which can be translocated within the plant.

The cyanogenic syndrome in rubber tree Hevea brasiliensis: tissue-damage-dependent activation of linamarase and hydroxynitrile lyase accelerates hydrogen cyanide release.

Background and Aims The release of hydrogen cyanide (HCN) from injured plant tissue affects multiple ecological interactions. Plant-derived HCN can act as a defence against herbivores and also plays an important role in plant–pathogen interactions. Crucial for activity as a feeding deterrent is the amount of HCN generated per unit time, referred to as cyanogenic capacity (HCNc). Strong intraspecific variation in HCNc has been observed among cyanogenic plants. This variation, in addition to genotypic variability (e.g. in Trifolium repens), can result from modifications in the expression level of the enzymes involved in either cyanogenic precursor formation or HCN release (as seen in Sorghum bicolor and Phaseolus lunatus). Thus, a modification or modulation of HCNc in reaction to the environment can only be achieved from one to the next generation when under genetic control and within days or hours when transcriptional regulations are involved. In the present study, it is shown that in rubber tree (Hevea brasiliensis) HCNc is modulated by post-translational activity regulation of the key enzymes for cyanide release. Methods Linamarase (LIN) and hydroxynitrile lyase (HNL) activity was determined by colorimetric assays utilizing dissociation of the substrates p-nitrophenyl-β-d-glucopyranoside and acetone cyanohydrin, respectively. Key Results In rubber tree leaves, LIN and HNL show up to ten-fold increased activity in response to tissue damage. This enzyme activation occurs within seconds and results in accelerated HCN formation. It is restricted to the damaged leaf area and depends on the severity of tissue damage. Conclusions LIN and HNL activation (in contrast to genetic and transcriptional regulations) allows an immediate, local and damage type-dependent modulation of the cyanogenic response. Accordingly, this post-translational activation plays a decisive role in the defence of H. brasiliensis against herbivores as well as pathogens and may allow more flexible reactions in response to these different antagonists.

Changes in cyanogenic glucoside content in seeds and seedlings of Hevea species

Abstract The content of cyanogenic glucosides in seeds of several Hevea species was investigated. Whereas in all seeds tested large quantities of the cyanogenic monoglucoside, linamarin, were detectable, lotaustralin, which occurs in much lower concentrations, did not occur in every individual seed. After storage, the cyanogenic diglucosides, linustatin and neolinustatin, were also detectable in Hevea seeds, implying that the monoglucosides, linamarin and lotaustralin, are glucosylated to yield the respective diglucosides, linustatin and neolinustatin. The content of cyanogenic glucosides was also monitored during germination and seedling development of several Hevea species. Depending on the species investigated, and even on the varieties of one species, the patterns of developmental changes in HCN-potential were quite different. Whereas in ‘wild type’ H. brasiliensis seedlings the HCN-potential decreased to less than 15% of the initial content, it remains nearly constant in those of variety FX 25. However, during seedling development of H. benthamiana, H. camargoana and H. pauciflora, the HCN-potential first increased and later decreased. These changes depend on two contrary processes, conversion of cyanogenic glucosides to non-cyanogenic compounds and de novo synthesis of cyanogens. Variations result from quantitative differences in consumption and de novo synthesis of cyanogenic glucosides as well as from differences in the times when these two processes are initiated.