Discrimination of sunflower volatiles by the red sunflower seed weevil

Five principle monoterpenoid and other constituent volatile chemicals of sunflower heads were combined to resemble two lines of sunflower (Helianthus annuus L.): one U.S.D.A. standard line and one French line which was poorly visited by insects (Etievant et al., 1984). Field trials of attraction to red sunflower seed weevils (Smicronyx fulvus Le Conte, Coleoptera: Curculionidae) showed that one was clearly preferred over the other. The more attractive mixture contained -pinene, -pinene, limonene, camphene and bornyl acetate in a ratio resembling that of Flath et al. (1985) rather than that described by Etievant et al. (1984). One or two volatiles were deleted from the optimal blend but only mixtures of five volatiles showed the highest attraction. Substitution of sabinene, another volatile prominent in sunflower, for one of the five in the optimal blend also decreased attraction of seed weevils. When the monoterpenoid components and green leaf volatiles in the traps resembled the ratios of most of the prominent volatiles of sunflower, attraction was significantly greater than controls.     

Proteomic analysis of oil body membrane proteins accompanying the onset of desiccation phase during sunflower seed development

A noteworthy metabolic signature accompanying oil body (OB) biogenesis during oilseed development is associated with the modulation of the oil body membranes proteins. Present work focuses on 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-based analysis of the temporal changes in the OB membrane proteins analyzed by LC-MS/MS accompanying the onset of desiccation (20–30 d after anthesis; DAA) in the developing seeds of sunflower (Helianthus annuus L.). Protein spots unique to 20–30 DAA stages were picked up from 2-D gels for identification and the identified proteins were categorized into 7 functional classes. These include proteins involved in energy metabolism, reactive oxygen scavenging, proteolysis and protein turnover, signaling, oleosin and oil body biogenesis-associated proteins, desiccation and cytoskeleton. At 30 DAA stage, exclusive expressions of enzymes belonging to energy metabolism, desiccation and cytoskeleton were evident which indicated an increase in the metabolic and enzymatic activity in the cells at this stage of seed development (seed filling). Increased expression of cruciferina-like protein and dehydrin at 30 DAA stage marks the onset of desiccation. The data has been analyzed and discussed to highlight desiccation stage-associated metabolic events during oilseed development.     

Purification and characterisation of an inhibitor of a cathepsin B-like proteinase from sunflower seed

Cathepsin B is a vitally important enzyme in various physiological processes and in tumor invasion and metastasis. A cathepsin B inhibitor, HCB-SunI, was identified and purified from sunflower seeds, Helianthus annuus, using ammonium sulfate precipitation and two steps of conventional chromatography. The molecular mass of HCB-SunI was estimated to be 12 kDa by SDS-PAGE and 12.32 kDa by MALDI TOF MS. Its N-terminal amino acid sequence was determined to be: PYGGGGTESG. HCB-SunI not only inhibited Helicoverpa cathepsin B (HCB) but also decreased the growth of HeLa and glioma cells by 7 ～ 27% and 6 ～ 22%, respectively, when the cells were grown in a final concentration of 0.002 ～ 0.008 μM inhibitor.     

Changes in the amounts of chlorogenic and caffeic acids during tobacco seed maturation

During maturation of tobacco seed, the amount of chlorogenic acid rapidly increased between days 7 and 11 after anthesis and then decreased until day 15. Chlorogenic acid was not detected in fully mature seed. On the other hand, caffeic acid appeared from day 15, reached a maximum at day 21, and rapidly disappeared by day 25 although the amount was always low in comparison with that of chlorogenic acid. Changes in dry wt, water content, testa color and germination of tobacco seeds during maturation were also investigated. The results suggested that there were rapid biochemical changes in seed between days 11 and 15 after anthesis.     

Phospholipids are present in extracellular fluids of imbibing sunflower seeds and are modulated by hormonal treatments

Phospholipids are well known messengers involved in developmental and stress responses mediating intracellular signalling. It has been hypothesized that phospholipids exist which could participate in intercellular communication events through the apoplast of sunflower (Helianthus annuus) seeds. Here it is shown that extracellular washing fluids (EWFs) obtained from seeds imbibed for 2 h contain diverse phospholipids. Lipid profiling by electrospray ionization tandem mass spectrometry revealed that the EWFs have a particular composition, with phosphatidic acid (PA) and phosphatidylinositol (PI) being the major phospholipids. These profiles are clearly distinct from those of seed extract (SE), and comparative SDS-PAGE of EWF and SE, followed by intracellular and plasma membrane marker analyses, allowed a significant contamination of the EWF to be discarded. Treatment of the seeds with 100 microM jasmonic acid (JA) induces changes in the profile of EWF phospholipids, leading to a decrease in PI content, while the accumulation of phosphatidylinositol 4-phosphate (PI4P) and specific PA species is observed. On the other hand, the EWF from seeds subjected to 50 microM abscisic acid (ABA) treatment exhibit an increase in PA and phosphatidylglycerol levels. To our knowledge, this is the first report on the existence of phospholipids as extracellular components of seeds. Moreover, the modulation of PA, PI, and PI4P levels by hormonal treatments further suggests their contribution to intercellular communication in planta.     

Ageing-induced changes in glutathione system of sunflower seeds

The glutathione system is thought to be involved in defence mechanisms present in plant tissues. The efficacy of this system was evaluated in large seeds of sunflower ( Helianthus annuus L. cv. Peredovik) in response to accelerated ageing (43°C/75% relative humidity from 1 to 11 days). Differences between the embryo axis and cotyledons in relation to the glutathione system were also investigated. Additionally, lipid peroxidation was determined by measuring the malondialdehyde (MDA) content. All assays were performed using dry seeds and seeds subsequently hydrated by imbibition in distilled water for 12 h at 25°C. Accelerated ageing caused a marked decrease in seed viability, accompanied by an increase in mean germination time. There were no changes in total glutathione in dry seeds. However, the distribution in its reduced (GSH) and oxidized (GSSG) forms revealed that ageing produced a slow conversion from GSH to GSSG. As the ageing period increased, this effect was accompanied by a decrease in glutathione reductase (GR, EC 1.6.4.2) activity. The results also indicated that the GSH system exerts a different response in the embryo axis as compared with the cotyledon: (1) the GSH levels decreased less in the cotyledons than in axes of aged seeds, and (2) the GSSG level in cotyledons was independent of ageing, while its amount increased in aged embryo axes. These different responses, in conjunction with the lower MDA levels in large as compared with small seeds, indicate a possible protective role of the reserve lipids. The efficacy of the GSH system in aged seeds was associated with seed viability, as revealed by multiple regression analysis. Upon imbibition, aged seeds were able to restore their GSH levels, reaching values approximating those of unaged seeds.     

Influence of feeding whole sunflower seed and extruded linseed on production of dairy cows,rumen and plasma constituents,and fatty acid composition of milk

Holstein cows were fed total mixed rations (TMR) supplemented with protected palm fat (PPF), whole sunflower seed (WSS) or extruded linseed (ELS) for 100 days. Percentage of dietary crude fat was 5.3, 5.1 and 5.1, respectively. Diet had no (p > 0.05) effect on feed intake, milk yield or milk protein content. Percentage of milk fat and yield of fat – corrected milk were significantly increased when diets were supplemented with WSS and ELS. Feeding PPF resulted in the lowest (p < 0.05) ruminal concentration of volatile fatty acids. No significant dietary effect on plasma characteristics was observed. Concentration of polyunsaturated fatty acids (PUFA) was higher (p < 0.05), and PUFA n-6/n-3 ratio lower (p < 0.05), in the milk fat from cows fed ELS compared to WSS. Supplementation of TMR with oilseeds compared to PPF increased the content of CLA in milk fat (p < 0.005) and decreased its atherogenicity, primarily due to a significant reduction of palmitic acid concentration. Both oilseeds significantly improved the spreadability index of manufactured butter. ELS, but not WSS, increased the susceptibility of milk fat to oxidation (p < 0.05). It can be concluded that feeding of oilseeds to dairy cows improved nutritional quality of milk fat, with supplementation with ELS producing an even more desirable milk fatty acid profile than WSS supplementation.     

Function of the ascorbate-glutathione cycle in aged sunflower seeds

The function of the ascorbate-glutathione (AsA/GSH) cycle was analyzed in seeds of sunflower ( Helianthus annuus L. cv. Peredovik) subjected to accelerated ageing at 43°C and 75% relative humidity for 1 to 11 days. The study was performed using dry seeds and seeds hydrated by imbibition in distilled water for 4 h at 25 °C. Lipid peroxidation was also determined by measuring the malondialdehyde (MDA) level. As the ageing period increased, a progressive loss of seed viability became increasingly evident. Even though high levels of MDA were delected, the MDA level did not change during accelerated ageing, suggesting that lipid peroxidation might occur to some extent. The study of the ascorbate/glutathione (AsA/GSH) cycle revealed that the GSH system is the major detoxifying mechanism in both dry and imbibed sunflower seeds. The GSH system is mainly located in the embryo, and its protective role is mediated by reactions that consume the GSH pool and, thereby, minimize the increase of the oxidized form (GSSG). Seed imbibition activates cellular metabolism and allows some antioxidant enzymes like glutathione reductase (EC 1,6,4,2) to act upon toxic agents. These reactions provide a reducing status, so that repair of damage becomes possible. However, prolonged ageing conditions (11 days) result in an irreversible damage, as evidenced by the appearance of dead seeds when the germination period ended. Multiple regression analysis revealed the effectiveness of the GSH system in aged seeds, especially upon imbibition and until the AsA/GSH cycle became completely functional.     

The Effects of Drying Temperature on the Extractability of Metals from Dredged Sediments

Dredged sediments may contain considerable amounts of metallic and/or organic pollutants. The risk of metal transfer from sediment deposits to soils can be evaluated using chemical extraction procedures; however, the temperatures at which samples are dried before metal extraction vary widely from one study to another. This led us to investigate the impact of drying temperature on the extractability of metals from dredged sediments. First, water-, CaCl ₂ - and DTPA-extractions were performed on 12 dredged sediments collected in France, with analyses being carried out on both raw (i.e. not dried) samples and on samples dried at 105°C. Higher extractable Cd and Zn contents were recorded for the samples dried at 105°C than for the raw samples. In order to assess the effect of drying temperature on metal extractability, we selected one sediment and carried out CaCl ₂ -, HNO ₃ - and DTPA-extractions on the raw sample and on samples dried at three different temperatures. In general, increasing the drying temperature led to an increase in the extractability of Cr and Cu for all three extraction methods. The CaCl ₂ -extractability of Ni and Zn decreased as drying temperature increased, except for the sample dried at 105°C. The HNO ₃ - and DTPA-extractabilities of these two metals were highest when the sediments were air-dried and then decreased with increasing drying temperature. Similar behavior was observed for Cd, except in the case of HNO ₃ -extractability. As the aim of chemical analysis using selective extraction is to measure the bio-available fractions of metals in soils and sediments, we suggest that extraction should be carried out on raw samples in order to simulate plant growth conditions.     

Synthesis and antioxidant properties of dendritic polyphenols

Three dendritic polyphenols (generation 1) were synthesized: a syringaldehyde-based dendrimer (1), a vanillin-based dendrimer (2), and an iodinated vanillin-based dendrimer (3). They all showed strong antioxidant activity according to the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical assay. The syringaldehyde dendrimer was twice and 10 times stronger than quercetin and Trolox, respectively. The vanillin-based dendrimer and its more hydrophobic iodinated derivative were also more potent antioxidants than quercetin and Trolox. The DPPH order of potency was 1 > 2, 3 > quercetin > Trolox. All three dendrimers also protected human LDL from free radical attack in a dose-dependent manner. Their order of free radical scavenging was 1 > 3 > 2 > quercetin > Trolox. The increased hydrophobic nature of the iodinated derivative may have contributed to its better LDL protection than 2. Protection of linoleic acid oxidation was studied by the β-carotene–linoleate assay. Dendrimer 1 was clearly superior to the other antioxidants in protecting the fatty acid. In case of DNA protection against free radical damage, the order of activity was 1 > quercetin > 2 > 3, Trolox. Pro-oxidant effect on copper-induced DNA oxidation showed the following order: quercetin, Trolox > 1 > 2 > 3. Results of the study show that dendritic antioxidants, even at the generation 1 level, provide promising antioxidant properties for their potential use as drug candidates for diseases associated with oxidative stress.     

Preparation of capacitor’s electrode from sunflower seed shell

Series of nanoporous carbons are prepared from sunflower seed shell (SSS) by two different strategies and used as electrode material for electrochemical double-layer capacitor (EDLC). The surface area and pore-structure of the nanoporous carbons are characterized intensively using N₂ adsorption technique. The results show that the pore-structure of the carbons is closely related to activation temperature and dosage of KOH. Electrochemical measurements show that the carbons made by impregnation-activation process have better capacitive behavior and higher capacitance retention ratio at high drain current than the carbons made by carbonization-activation process, which is due to that there are abundant macroscopic pores and less interior micropore surface in the texture of the former. More importantly, the capacitive performances of these carbons are much better than ordered mesoporous carbons and commercial wood-based active carbon, thus highlighting the success of preparing high performance electrode material for EDLC from SSS.     

Transgenic Trifolium repens with foliage accumulating the high sulphur protein,sunflower seed albumin

With the aim of increasing the rumen-protected level of the sulphur amino acids cysteine and methionine in Trifolium repens, we introduced the coding sequence of the sunflower seed albumin (SSA) into T. repens by Agrobacterium tumefaciens-mediated transformation. The SSA gene was modified such that the protein would be localised to the endoplasmic reticulum (ER). Four different T-DNA constructions all containing the SSA gene driven by either the promoter of a gene encoding the small subunit of ribulose bisphosphate carboxylase (Rubisco) from Arabidopsis thaliana (A _ssu), the promoter of the gene encoding the small subunit of Rubisco of Medicago sativa (L _ssu), or the Cauliflower Mosaic Virus 35S promoter (CaMV35S), were transferred to T. repens cv. Haifa. Transgenic T ₀-plants and inter-transgenic hybrids were analysed for the level of SSA accumulation in the leaves by western blotting. The highest observed level of SSA accumulation was 0.1% of total extractable leaf protein. We observed that the promoter had a substantive effect on the level of SSA accumulation with A _ssu>CaMV35S>L _ssu. Results from the inter-transgenic hybrids showed that the capacity to synthesise SSA was inherited. However the level of SSA accumulation in the leaves generally appears not to be additive with extra transgenic loci. During this work, we attempted to improve the efficiency of A. tumefaciens-mediated transformation of T. repens using the SAAT-method (Sonication Assisted Agrobacterium-mediated Transformation) on cotyledons of T. repens. T-DNA transfer was in general not enhanced by sonication compared to traditional A. tumefaciens-mediated transformation. Furthermore, Southern blot analyses of plants regenerated from the same cotyledon after A. tumefaciens treatment and under selection, indicated that multiple shoots were usually derived from the same transformation event. We concluded from these results that only one plant from each A. tumefaciens-treated cotyledon should be taken to avoid transgenic clones.     

Identification of free radical species derived from caffeic acid and related polyphenols

Polyphenols are widely distributed in various fruits, vegetables and seasonings. It is well known that they have several physiological effects due to their antioxidative activities. Their activities depend on structural characteristics that favour the formation of their corresponding stable radicals. During the examination at which pH values, the polyphenol radicals are stabilized, we confirmed that polyphenol radicals were stabilized in NaHCO₃/Na₂CO₃ buffer (pH 10) rather than in physiological pH region. Then, we measured electron spin resonance (ESR) spectra at pH 10 to examine the characteristics of free radical species derived from caffeic acid (CA) with an unsaturated side chain, dihydrocaffeic acid (DCA) with a saturated side chain, chlorogenic acid (ChA) and rosmarinic acid (RA). In analyzing the radical structures, ESR simulation, determinations of macroscopic and microscopic acid dissociation constants and molecular orbital (MO) calculation were performed. In CA, the monophenolate forms were assumed to participate in the formation of free radical species, while in DCA, the diphenol form and the monophenolate forms were presumed to contribute to the formation of free radical species. On the basis of the results, we propose the possible structures of the free radical species formed from polyphenols under alkaline conditions.     

Identification of free radical species derived from caffeic acid and related polyphenols

Polyphenols are widely distributed in various fruits, vegetables and seasonings. It is well known that they have several physiological effects due to their antioxidative activities. Their activities depend on structural characteristics that favour the formation of their corresponding stable radicals. During the examination at which pH values, the polyphenol radicals are stabilized, we confirmed that polyphenol radicals were stabilized in NaHCO₃/Na₂CO₃ buffer (pH 10) rather than in physiological pH region. Then, we measured electron spin resonance (ESR) spectra at pH 10 to examine the characteristics of free radical species derived from caffeic acid (CA) with an unsaturated side chain, dihydrocaffeic acid (DCA) with a saturated side chain, chlorogenic acid (ChA) and rosmarinic acid (RA). In analyzing the radical structures, ESR simulation, determinations of macroscopic and microscopic acid dissociation constants and molecular orbital (MO) calculation were performed. In CA, the monophenolate forms were assumed to participate in the formation of free radical species, while in DCA, the diphenol form and the monophenolate forms were presumed to contribute to the formation of free radical species. On the basis of the results, we propose the possible structures of the free radical species formed from polyphenols under alkaline conditions.     

不同种源青檀种子的营养成分及种子活力的差异

以采自安徽（ＡＨ）、山东（ＳＤ）及江苏（ＪＳ）的６个青檀（ＰｔｅｒｏｃｅｌｔｉｓｔａｔａｒｉｎｏｗｉＭａｘｉｍ．）种源种子为材料,测定各种源种子营养成分含量及种子活力。６个种源种子的蛋白质、可溶性糖、淀粉及粗脂肪含量存在较大差异。在种子品质上,６个种源的种子千粒重、浸泡液电导率及简化种子活力指数（ＳＶＩＳ）也存在显著差异。种子中各营养成分含量的高低与ＳＶＩＳ的大小密切相关,复相关系数达０８２２３,以蛋白质含量对ＳＶＩＳ的影响最大,淀粉和粗脂肪含量影响次之,可溶性糖含量影响最小。采用模糊聚类分析法,以种子千粒重、电导率及ＳＶＩＳ为评价指标,可将６个种源的种子品质划分为三大类：第一类包括ＳＤ１、ＡＨ１和ＪＳ１３个种源,种子品质较好;第二类仅为ＡＨ３种源,品质中等;第三类包括ＡＨ２和ＳＤ２２个种源,品质较差。     

Immunological estimation of growth regulator distribution in phototropically reacting sunflower seedlings

The distribution of immunoassayable xanthoxin (XA), abscisic acid (ABA) and indole-3-acetic acid (IAA) in all parts of sunflower ( Helianthus annuus L.) seedlings was determined. During the course of phototropic curvature, including the lag phase (5 min), the distribution of these growth regulators was analyzed in the illuminated and shaded side of the hypocotyl, as well as in the peripheral and central tissues. All three growth regulators showed no detectable asymmetries between the illuminated and shaded hypocotyl halves during the lag phase and early phototropic curvature. Also, no indication for an exchange of XA, ABA or IAA between the peripheral and central tissues was observed. Partial removal of the peripheral cell layers revealed that changes in the growth properties of this tissue, preferentially at the illuminated side of the hypocotyl, are required for the phototropic reaction. Complete removal of the peripheral cell layers abolishes the phototropic response. In dark-incubated, green sunflower seedlings, the loss of sensitivity to phototropic stimulation is correlated with decreasing levels of IAA immunoreactivity, whereas no changes in the levels of ABA- and XA immunoreactivity were recorded. The findings are discussed with respect to the involvement of ABA, XA and IAA in phototropic reactions of green dicotyledonous shoots.     

Enhanced sunflower oil utilization and gamma-linolenic acid production by Mucor circinelloides f.circinelloides CBS 108.16 in the presence of acetate

World Journal of Microbiology and Biotechnology -     

Effect of soil chloride level on cadmium concentration in sunflower kernels

Understanding soil factors related to cadmium (Cd) uptake and accumulation in plants is important for development of agronomic technologies, and breeding strategy to produce low Cd crops. The objective of the study was to examine the effect of soluble salts (chloride and sulfate) and other soil factors on the Cd concentration in sunflower (Helianthus annuus L.) kernels. Commercial nonoilseed hybrid kernels and soils were sampled from 22 farmer's production fields in North Dakota and Minnesota. The sites sampled included saline and nonsaline variants from 7 soil series. Soils were sampled at four depths. Relationships between kernel Cd level and soil physical and chemical characteristics were examined. The soil pH covered a narrow range (7.3–8.1) at these sampled sites. Regression analysis showed that there was no correlation between kernel Cd and soil pH at any depth. The kernel Cd level was highly correlated with DTPA-extractable Cd in all 4 depths, and with clay content in sub-soils. Soil chloride and sulfate concentrations varied among soil series and within soil series. The absence of a statistically significant effect of soil sulfate level on kernel Cd concentration, indicated that soil sulfate levels did not affect Cd uptake by sunflower plants. However, soil chloride levels in sub-soil were correlated with kernel Cd. The most important soil factor was DTPA-extractable Cd. When chloride was included in the multiple regression equations, R square (R²) values improved significantly. These results demonstrate that soil chloride concentration is another important factor related to Cd uptake in sunflower plants.     

Continuous lipase-catalyzed production of esters from crude high-oleic sunflower oil

The objective of this work was to develop an economically relevant enzymatic process of butyl ester production using crude high-oleic sunflower oil. Novozym 435, a non-regiospecific biocatalyst, provided the best compromise between activity and butyl-ester yield. The inhibition caused by the presence of phopholipids in crude oil was eliminated by using tert-butanol. It demonstrates the key role of the medium polarity in order to insure the stability of a process. Initial substrate concentrations and their molar ratio were optimized in a continuous packed-bed reactor to maximize product yield and productivity. The best compromise was obtained for an initial oil concentration of 500 mM and a molar ratio of 5. It enabled a high productivity of 13.8 tons year⁻¹ kg Novozym 435⁻¹ with a butyl-ester purity of 96.5% to be obtained. Experiments with the continuous reactor were performed over 50 days without any loss of enzyme activity.