Sulfur and nitrogen fertility affects flavour of field-grown onions

Although glasshouse studies have conclusively demonstrated that S nutrition can affect onion (Allium cepaL.) pungency this has been rarely observed in field-based studies due to difficulties in controlling S nutrition and lack of efficient methods for measurement of flavour bioactives. We have developed a rapid automated method for determination of onion lachrymatory factor ((Z, E)-thiopropanal-S-oxide; LF) based on juice extraction into dichloromethane and gas chromatography (GC) analysis with flame photometric detection. We evaluated this in a field trial of a mild (cv. ‘Encore’) and a pungent (cv. ‘Kojak’) onion cultivar grown on a low S soil with and without S addition, under high or low N treatments. No treatments significantly affected bulb fresh weight but S fertilisation significantly increased bulb total S, sulfate, pungency, LF and flavour precursor levels in both varieties. Analysis of bulb flavour precursors by HPLC confirmed that juice LF levels paralleled levels of the flavour precursor S-1-propenyl cysteine sulfoxide. The pungent cultivar also exhibited significant N main effects on bulb LF, total S and sulfate. We also assayed the key S-assimilatory enzyme, APS reductase (APR) in leaves before and during bulbing. Specific activities in the range of 1 to 11 nmol mg⁻¹·min⁻¹ were observed in youngest leaves, but only the milder cultivar exhibited significant stage × N × S effects. These findings suggest that sulfur metabolism of mild and pungent onions respond differently to N fertility, and that GC of LF is practical for field-based studies of onion flavour.     

Effect of nitrogen levels of two cherry tomato cultivars on development,preference and honeydew production of Trialeurodes vaporariorum (Hemiptera: Aleyrodidae)

Two cherry tomato plant cultivars (Lycopersicon esculentum Miller, cultivars ‘Koko’ and ‘Pepe’) were supplied with high (395 ppm), medium (266 ppm) and low (199 ppm) concentrations of nitrogen to determine the influence of nitrogen fertilization on development, cultivar preference and honeydew production by greenhouse whiteflies, Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae). The nitrogen, protein, and chlorophyll contents of tomato leaves were higher in the high nitrogen supplied plants than in the medium or low nitrogen supplied plants, but the sugar content showed an inverse relationship. The developmental times of eggs and nymphs decreased as the nitrogen concentrations increased in both cultivars. The preference of T. vaporariorum was compared by counting the number of eggs deposited on leaves in choice and non-choice tests. In the non-choice test, no significant nitrogen treatment effects were observed but the upper plant stratum was preferred for egg laying. In the choice test, there were significant main effects of cultivar and nitrogen concentration. T. vaporariorum laid eggs more on leaves of plants with higher nitrogen at the upper stratum. In both experiments, T. vaporariorum preferred the ‘Koko’ cultivar to the ‘Pepe’ cultivar. The honeydew production of T. vaporariorum nymphs increased with decreasing nitrogen treatment concentrations. The largest honeydew production was detected in the ‘Pepe’ cultivar grown at low nitrogen concentration. It is concluded that cultivar ‘Pepe’ had an advantage over ‘Koko’ in term of T. vaporariorum management program in tomato greenhouses.     

Genotypic variation in sulphur assimilation and metabolism of onion (Allium cepa L.). II: Characterisation of ATP sulphurylase activity

To investigate the regulation of sulphur (S)-assimilation in onion further at the biochemical level, the pungent cultivar W202A and the milder cultivar Texas Grano 438 PVP (TG) have been grown in S-sufficient (S⁺; 4 meq S⁻¹) or S-deficient (S⁻; 0.1 meq S⁻¹) growth conditions, and tissues excised at the seedling stage (pre-bulbing; ca. 10-weeks-old) and at the mature stage (bulbing; ca. 16-weeks-old). S-supply negatively influenced adenosine-5′-phosphosulphate (APS) reductase (APR) enzyme activity in both cultivars at bulbing only, and a higher abundance of APR was observed in both cultivars at bulbing in response to low S-supply. In contrast, S-supply significantly influenced ATP sulphurylase (ATPS) activity in leaf tissues of W202A only, and only at bulbing, while an increase in abundance in response to high S-supply was observed for both cultivars at bulbing. To investigate the regulation of the ATPS enzyme activity and accumulation further, activity was shown to decrease significantly in roots at bulbing in the S-deficient treatment in both cultivars, a difference that was only supported by western analyses in W202A. Phylogenetic analysis revealed that AcATPS1 groups in a broad monocot clade with the closest sequences identified in Sorghum bicolour, Zea mays and Oryza sativa, but with some support for a divergence of AcATPS1. Detection of ATPS in leaf extracts after two dimensional gel electrophoresis (2-DE) revealed that the protein may undergo post-translational modification with a differential pattern of ATPS accumulation detected in both cultivars over the developmental progression from the seedling to the bulbing stage. Treatment of leaf extracts of W202A to dephosphorylate proteins resulted in the loss of immuno-recognised ATPS spots after 2-DE separation, although enzyme activity was not influenced. These results are discussed in terms of the tiers of control that operate at the biochemical level in the reductive S-assimilation pathway in a S-accumulating species particularly during the high-S-demanding bulbing stage.     

Influence of bacterial streak disease development on changes in phenolics,soluble amino acids and carbohydrates of rice leaves

The healthy leaves of rice cultivar ‘BJ 1’ resistant to bacterial leaf streak pathogen (Xanthomonas translucens f. sp.oryzicola) contained higher quantities of total phenolic compounds, reducing and nonreducing sugars than the susceptible cultivar ’IR 8’, while the leaves of cultivar ’IR 8’ possessed larger concentration of total soluble amino acids than the resistant cultivar ’BJ 1’. In the leaves of cultivar ‘BJ 1’, the disease development caused an initial decrease in the concentration of phenols followed by an increase at later stages. As a result of inoculation, soluble carbohydrates and amino acids generally decreased in the leaves of resistant cultivar ‘BJ 1’, in contrast to an increase in their concentration in the leaves of cultivar ‘IR 8’.     

Targeted knock-out of a gene encoding sulfite reductase in the moss Physcomitrella patens affects gametophytic and sporophytic development

A key step in sulfate assimilation into cysteine is the reduction of sulfite to sulfide by sulfite reductase (SiR). This enzyme is encoded by three genes in the moss Physcomitrella patens. To obtain a first insight into the roles of the individual isoforms, we deleted the gene encoding the SiR1 isoform in P. patens by homologous recombination and subsequently analysed the ΔSiR1 mutants. While ΔSiR1 mutants showed no obvious alteration in sulfur metabolism, their regeneration from protoplasts and their ability to produce mature spores was significantly affected, highlighting an unexpected link between moss sulfate assimilation and development, that is yet to be characterized.     

Molecular cloning and characterization of a gene regulating flowering time from Alfalfa (Medicago sativa L.)

Genes that regulate flowering time play crucial roles in plant development and biomass formation. Based on the cDNA sequence of Medicago truncatula (accession no. AY690425), the LFY gene of alfalfa was cloned. Sequence similarity analysis revealed high homology with FLO/LFY family genes of other plants. When fused to the green fluorescent protein, MsLFY protein was localized in the nucleus of onion (Allium cepa L.) epidermal cells. The RT-qPCR analysis of MsLFY expression patterns showed that the expression of MsLFY gene was at a low level in roots, stems, leaves and pods, and the expression level in floral buds was the highest. The expression of MsLFY was induced by GA₃ and long photoperiod. Plant expression vector was constructed and transformed into Arabidopsis by the agrobacterium-mediated methods. PCR amplification with the transgenic Arabidopsis genome DNA indicated that MsLFY gene had integrated in Arabidopsis genome. Overexpression of MsLFY specifically caused early flowering under long day conditions compared with non-transgenic plants. These results indicated MsLFY played roles in promoting flowering time.     

Neutron scattering maps the higher-order assembly of NADPH-dependent assimilatory sulfite reductase

Precursor molecules for biomass incorporation must be imported into cells and made available to the molecular machines that build the cell. Sulfur-containing macromolecules require that sulfur be in its S^2? oxidation state before assimilation into amino acids, cofactors, and vitamins that are essential to organisms throughout the biosphere. In α-proteobacteria, NADPH-dependent assimilatory sulfite reductase (SiR) performs the final six-electron reduction of sulfur. SiR is a dodecameric oxidoreductase composed of an octameric flavoprotein reductase (SiRFP) and four hemoprotein metalloenzyme oxidases (SiRHPs). SiR performs the electron transfer reduction reaction to produce sulfide from sulfite through coordinated domain movements and subunit interactions without release of partially reduced intermediates. Efforts to understand the electron transfer mechanism responsible for SiR’s efficiency are confounded by structural heterogeneity arising from intrinsically disordered regions throughout its complex, including the flexible linker joining SiRFP’s flavin-binding domains. As a result, high-resolution structures of SiR dodecamer and its subcomplexes are unknown, leaving a gap in the fundamental understanding of how SiR performs this uniquely large-volume electron transfer reaction. Here, we use deuterium labeling, in vitro reconstitution, analytical ultracentrifugation (AUC), small-angle neutron scattering (SANS), and neutron contrast variation (NCV) to observe the relative subunit positions within SiR’s higher-order assembly. AUC and SANS reveal SiR to be a flexible dodecamer and confirm the mismatched SiRFP and SiRHP subunit stoichiometry. NCV shows that the complex is asymmetric, with SiRHP on the periphery of the complex and the centers of mass between SiRFP and SiRHP components over 100 Å apart. SiRFP undergoes compaction upon assembly into SiR’s dodecamer and SiRHP adopts multiple positions in the complex. The resulting map of SiR’s higher-order structure supports a cis/trans mechanism for electron transfer between domains of reductase subunits as well as between tightly bound or transiently interacting reductase and oxidase subunits.     

Changes in lipid composition, water relations and gas exchange in leaves of two young ‘Chemlali’ and ‘Chetoui’ olive trees in response to water stress

The comparative responses of two young olive trees (Olea europaea L. ‘Chemlali’ and ‘Chetoui’) to drought stress were investigated during 1 month. Three-month-old own-rooted plants were subjected to two irrigation treatments: WW (well watered plants that were irrigated with fresh water to maintain a soil water content close to field capacity), and WS (water stressed plants by withholding water). Leaf water potential, gas exchange and leaf lipid composition were studied. ‘Chemlali’ was able to maintain higher leaf CO₂ assimilation rate and leaf stomatal conductance throughout the drought cycle compared to ‘Chetoui’. Water stress induced a larger decrease in the total lipid content in ‘Chetoui’ than in ‘Chemlali’. Interestingly, the constitution of different lipid classes was highly altered in ‘Chetoui’. Lipid changes in Chemlali, a drought tolerant cultivar, revealed more stability of its cellular membranes to drought stress as compared to the drought susceptible olive cultivar, Chétoui. Furthermore, in comparison to the controls, drought stressed plants showed an increase in the degree of unsaturation of leaf lipids in the two olive cultivars. Moreover, the results observed in Chemlali showed that besides changes in lipids composition this cultivar may have an efficient defence strategy which can be related on antioxidative production against oxidative stress.     

Interaction of sulfate assimilation with nitrate assimilation as a function of nutrient status and enzymatic co-regulation inbrassica juncea roots

The interaction of sulfate assimilation with nitrate assimilation inBrassica juncea roots was analyzed by monitoring the regulation of ATP sulfurylase (AS), adenosine-5’-phosphosulfate reductase (AR), sulfite reductase (SiR), and nitrite reductase (NiR). Depending on the status of sulfur and nitrogen nutrition, AS and AR activities and mRNA levels were increased by sulfate starvation but decreased by nitrate starvation. The activation of AS and AR by sulfate starvation was inhibited by sulfate/nitrate starvation. However, the rise in steady-state mRNA levels for AS and AR by sulfate starvation was not affected by sulfate/nitrate starvation. SiR gene expression was slightly activated by both sulfate starvation and sulfate/nitrate starvation, but was decreased by nitrate starvation. Although NiR gene expression was little affected by sulfate starvation, it was diminished significantly by either nitrate or nitrate/sulfate starvation. Cysteine (Cys) also decreased AS and AR activities and mRNA levels even when plants were simultaneously starved for sulfate; in contrast, both SiR and NiR gene expressions were only slightly, if at all, affected under the same conditions. This supports our conclusion that Cys, the end-product of sulfate assimilation, is the key regulatory signal. Moreover, SiR and NiR apparently are not the linking step in the co-regulation of sulfate and nitrate assimilation in plants.     

Essential oils from buds and leaves of two hazelnut (Corylus L.) cultivars with different resistance to filbert big bud mite (Phytoptus avellanae Nal.) and filbert aphid (Myzocallis coryli Goetze)

The aim of this study was to identify the allelopatic effect of the components of a mixture of essential oils (EO) contained in the buds and leaves of hazel (Corylus L.) on herbivores. We examined the effect of these compounds on the choice of plants of two different hazel cultivars by Phytoptus avellanae Nal. (filbert big bud mite) and Myzocallis coryli Goetze (filbert aphid), which are the most important pests of hazel in Poland and throughout the world. Our results show that plants of cv. ‘Mogulnus’ were more resistant than those of cv. ‘Barra’ to the feeding of mites and aphids in all study years. Using gas chromatography (GC) and GC/mass spectrometry methodology, we determined the qualitative and quantitative composition of EO in the buds and leaves of plants of these two hazel cultivars. The EO obtained from the analysed materials was a mixture of mono- and sesquiterpenes. The emission of volatile organic compounds from plants is known to repel or attract pests. The mixture of EO present in the hazel buds of cv. ‘Mogulus’, which is resistant to filbert big bud mite, was characterized by a high content of nerol, α-campholenol, methyl salicylate, spatulenol, β-caryophylene and δ-cadinene. In contrast, the leaves of this cultivar, colonized by filbert aphid but to a relatively small extent, contained greater quantities of nerol, α-campholenol, p-cymene, α-terpineol and germacrene D, than the leaves of cv. ‘Barra’, which is more accepted by aphids. However, the leaves of cv. ‘Barra’ were characterized by a considerably high content of menthol, limonene, isomenthone, methyl salicylate and L-menthone.