Functional characterization of FaCCD1: a carotenoid cleavage dioxygenase from strawberry involved in lutein degradation during fruit ripening

A gene encoding a carotenoid cleavage dioxygenase class 1 enzyme (FaCCD1) was identified among a strawberry fruit expressed sequence tag collection. The full-length cDNA was isolated, and the expression profiles along fruit receptacle development and ripening, determined by quantitative real time polymerase chain reaction, showed that FaCCD1 is a ripening-related gene that reaches its maximal level of expression in the red fully ripe stage. FaCCD1 was expressed in Escherichia coli, and the products formed by the recombinant protein through oxidative cleavage of carotenoids were identified by liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry analyses. The FaCCD1 protein cleaves zeaxanthin, lutein, and beta-apo-8'-carotenal in vitro. Although beta-carotene is not a good substrate for FaCCD1 in vitro, the expression of FaCCD1 in an engineered carotenoid-producing E. coli strain caused the degradation of beta-carotene in vivo. Additionally, the carotenoid profile in strawberry was analyzed by high-performance liquid chromatography-photodiode detection, and a correlation between the increase of the expression level of FaCCD1 during ripening and the decrease of the lutein content suggests that lutein could constitute the main natural substrate of FaCCD1 activity in vivo.     

Effects of humus content,farmyard manuring,and mineral‐N fertilization on yields and soil properties in a long‐term trial

Investigations carried out at Field F3 of the Halle long‐term fertilization trials using data from 1974 to 1983 showed that with adequate supply of mineral N‐fertilizer soil organic matter (SOM) had no significant effects of yield. Similarly enhanced SOM did not justify a reduction of mineral N (Stumpe et al., 2000). The studies presented here examine the effects of the SOM differences existing after the termination of those trials in 1986 up until 1997 (then mainly differences of hardly decomposable SOM) in comparison to farmyard manuring with enhanced mineral N application (3‐factor‐experiment). As with total SOM, hardly decomposable SOM did not directly affect yields. The effects of FYM treatment observed at lower mineral‐N levels were compensated for by enhanced mineral‐N supply. The direct effect of FYM (40 t ha^—1) corresponded to a mineral‐N supply of about 60 kg ha^—1 and the residual effect to about 20 kg ha^—1. The differences of the C‐content in the soil at the beginning of the present studies continued throughout the experimental period of 12 years. In addition, significant differentiation has been caused by FYM and N fertilization in comparison to unfertilized treatments. The major finding is that differences in SOM content do not lead to yield differences on physically good soils (chernozem‐like soils) if appropriate compensation by mineral‐N fertilization takes place.     

Changes in the subsoil of long‐term trials in Halle (Saale), Germany,caused by mineral fertilization

Four long‐term mineral fertilization trials (50 years) on a Haplic Phaeozem derived from sandy loess were conducted. Fertilization caused changes to the plough layer (0—25 cm) and to the undisturbed subsoil. Without lime, most remarkable acidification occurred in the plough layer, but only slight effects were evident in the subsoil. Where lime was applied, a considerable amount of Ca‐sulphate was formed. This leached out of the plough layer and accumulated throughout the profile, especially in dry years. Where fertilization was not adequate, there was a substantial contribution of K and P from the subsoil (mainly of its upper parts). Where the supply of these two elements continously exceeded their removal, a distinct increase of available K and P at depths of about 50 cm indicated transfer of both elements to the subsoil. Due to the high fixing capacity of both P and K in layers beneath 50 cm, leaching out of the rooting zone can be ignored. Mg and especially Na, applied with some of the fertilizers, were more mobile than K.     

Verbleib des 15N nach einmaliger Verabreichung von markiertem Düngerstickstoff im Dauerversuch Ewiger Roggenbau Halle

Fate of ¹⁵N after one single application of labelled N fertilizer in the long-term experiment Eternal Rye Cropping Halle To elucidate the fate of N in the long-term field experiment with continuous rye cropping in Halle, Germany (Haplic Phaeozem derived from sandy loess; mean annual precipitation: 466 mm) micro plots (9 m²) were established within the NPK main plot in 1993 and 1994 and the routine N dose (60 kg N ha^—1 a^—1) was once applied as ¹⁵N labelled fertilizer. In the subsequent four years each micro plot was analyzed for the ¹⁵N withdrawn with grain and straw and that remaining in the soil after harvest. Parallel to this, micro plots on the unfertilized main plot received a single dose of 5 kg ha^—1 of high-label ¹⁵N in order to achieve labelling of the annual N input from the air (˜40 kg ha^—1 a^—1) and to follow its fate. — In the NPK treatment about one third of the labelled fertilizer N applied (= *N) was taken up by the rye in the first year, while nearly half of it remained in the soil (0—100 cm), and 10—15% was not recovered (most likely lost via leaching). During the following three years the amount of *N remaining in the soil decreased from 32.7 kg ha^—1 to 21.5 kg ha^—1. However, less than half of the remobilized *N had been taken up by the crop. On the unfertilized plot the uptake of *N was lower at the end of the experiment compared to the NPK plot, while the loss exceeded the comparative figure correspondingly.     

Untersuchungen zur Charakterisierung des pflanzenverfügbaren Phosphats in Thüringer Carbonatböden

Investigations on the characterization of plant available phosphate in Thuringian calcareous soils Phosphate availability in Thuringian calcareous soils and its characterization by CAL‒, NaHCO₃-, H₂O- and CaCl₂-soil tests was investigated in laboratory and pot experiments. Soil CaCO₃ contents > 10% increase the pH value of CAL solution and thus decrease phosphate extractability. The increase of pH also causes an inadequate assessment of plant available phosphate by H₂O- and CaCl₂-soil tests. The CAL soluble P content of the soil corrected by the pH value of the extraction solution was most suitable for the forecast of P uptake of corn in a pot trial. From both parameters P availability indices for calcareous soils can be calculated which are comparable with those in soils containing < 5% CaCO₃. The equation, which still has to be verified in field experiments, reads as CAL-P_corrected = CAL-P_measured · (1 + 0.83·〈pH-value_{CAL solution} — 4.1〉).     

Anticancer Effects of Different Seaweeds on Human Colon and Breast Cancers

Seafoods and seaweeds represent some of the most important reservoirs of new therapeutic compounds for humans. Seaweed has been shown to have several biological activities, including anticancer activity. This review focuses on colorectal and breast cancers, which are major causes of cancer-related mortality in men and women. It also describes various compounds extracted from a range of seaweeds that have been shown to eradicate or slow the progression of cancer. Fucoidan extracted from the brown algae Fucus spp. has shown activity against both colorectal and breast cancers. Furthermore, we review the mechanisms through which these compounds can induce apoptosis in vitro and in vivo. By considering the ability of compounds present in seaweeds to act against colorectal and breast cancers, this review highlights the potential use of seaweeds as anticancer agents.     

Influence of a xylanase feed additive on Lactobacillus species in the jejunum of piglets

rRNA extracts from the jejunum of rearing piglets fed a wheat/rye based diet with or without a xylanase feed additive were hybridized against eight Lactobacillus spp. specific digoxigenin labelled oligonucleotide probes. Known concentrations of sample- or culture RNA extracts were used to calibrate and quantify hybridization signals from different sample extracts and hybridizations. The xylanase feed additive tended to increase total eubacterial rRNA in the terminal jejunum, probably a result of increased Lactobacillus spp. rRNA in the central and terminal jejunum. However, individual animal variations were high. L. reuteri was the major Lactobacillus species followed by L. acidophilus and L. mucosae. L. amylophilus and L. crispatus were not detected in all RNA extracts.The results of this study show that the xylanase feed additive modifies Lactobacillus spp. populations on the species level in piglets in addition to their well known effects on digesta viscosity and nutrient digestibility.The implications of these effects for the intestinal microbiota require further analysis.     

Influence of agronomic factors and extraction rate on the acrylamide contents in yeast-leavened breads

Because the impact of agronomical factors on bakery products quality is still an insufficiently studied field, acrylamide contents of breads produced from flours of nine wheat, two rye, and two spelt varieties harvested in 2003 and 2004 were investigated. It could be demonstrated that acrylamide content in bread strongly depends on the cultivar, with extremes differing by a factor of 5.4 due to marked differences in free asparagine and crude protein contents. Nitrogen fertilization also resulted in elevated amino acid and protein contents, thus increasing acrylamide levels from 10.6 to 55.6 mug/kg. Independent of fertilization, harvest year turned out to be another factor influencing acrylamide formation. Breads produced from 2003 flours showed significantly higher acrylamide contents than those of 2004, which was ascribed to favorable light and temperature conditions during the cultivation period, thus enhancing amino acid and protein contents. Sprouting of the grain also resulted in significantly higher acrylamide levels, which was attributed to elevated enzyme activities and the formation of precursors from protein and starch. Furthermore, bakery products made from flours with higher extraction rates were shown to contain higher acrylamide levels resulting from extracted free asparagine and protein from the aleuron layers of the cereal grain.     

Quantitation of the main constituents of some authentic sesame seed oils of different origin

This paper describes the composition of sesame seed oils obtained from seeds collected from five countries that are major suppliers of traded sesame seed oil. Oils were extracted from the seeds using small-scale industry pressing equipment and analyzed using standard methods for fatty acids, fatty acids in the triglyceride 2-position, tocopherols and tocotrienols, triglycerides, sterols, steradienes, and iodine value. Values for the composition of the sterols, triglycerides, fatty acids, iodine value, and tocopherol composition were generally in good agreement with the results published elsewhere. All of the oils from roasted seeds contained low levels of the sterol degradation products stigmasta-3,5-diene and campesta-3,5-diene, which were probably formed by dehydration of the parent sterols during roasting.     

Functional characterization of enone oxidoreductases from strawberry and tomato fruit

Fragaria x ananassa enone oxidoreductase (FaEO), earlier putatively assigned as quinone oxidoreductase, is a ripening-induced, negatively auxin-regulated enzyme that catalyzes the formation of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF), the key flavor compound in strawberry fruit by the reduction of the alpha,beta-unsaturated bond of the highly reactive precursor 4-hydroxy-5-methyl-2-methylene-3(2H)-furanone (HMMF). Here we show that recombinant FaEO does not reduce the double bond of straight-chain 2-alkenals or 2-alkenones but rather hydrogenates previously unknown HMMF derivatives substituted at the methylene functional group. The furanones were prepared from 4-hydroxy-5-methyl-3(2H)-furanone with a number of aldehydes and a ketone. The kinetic data for the newly synthesized aroma-active substrates and products are similar to the values obtained for an enone oxidoreductase from Arabidopsis thaliana catalyzing the alpha,beta-hydrogenation of 2-alkenals. HMMF, the substrate of FaEO that is formed during strawberry fruit ripening, was also detected in tomato and pineapple fruit by HPLC-ESI-MSn and became 13C-labeled when d-[6-13C]-glucose was applied to the fruits, which suggested that a similar HDMF biosynthetic pathway occurs in the different plant species. With a database search (http://ted.bti.cornell.edu/ and http://genet.imb.uq.edu.au/Pineapple/), we identified a tomato and pineapple expressed sequence tag that shows significant homology to FaEO. Solanum lycopersicon EO (SlEO) was cloned from cDNA, and the protein was expressed in Escherichia coli and purified. Biochemical studies confirmed the involvement of SlEO in the biosynthesis of HDMF in tomato fruit.