Use of megagametophyte storage proteins as markers of the genetic diversity in stone pine (Pinus pinea L.) in Andalucia, Spain

Stone pine (Pinus pinea L.) is a Mediterranean species of interest for the traditional use of his edible kernel. Its area of distribution includes all the Mediterranean area, including the Iberian Peninsula where this tree presents the most extensive natural stands. Since land reforestation has become a high-priority of European policy, there is an increased interest in the characterisation of the germplasm of this species. In this study, the globulins from the megagametophyte were used for analysis of the variability and genetic diversity in 20 populations from Andalucia. Because of the complex profile, these proteins were analysed as blocks, corresponding each to a different range of molecular weight. Polymorphisms were observed in each of the five blocks. The diversity data showed that these populations present a high degree of variation, which suggest that this technic could be useful to analyse of the genetic diversity in this species.     

Multiple domestications of the Mesoamerican gene pool of lima bean (Phaseolus lunatus L.): evidence from chloroplast DNA sequences

The Mesoamerican (MA) gene pool of lima bean (Phaseolus lunatus L.) is widely distributed from northern Mexico to northern Argentina in its wild forms and from the southern United States to the east coast of Brazil in its domesticated forms. This broad distribution and lack of wild accessions of the MA gene pool in many areas of its natural distribution has impeded determining its center of domestication and establishing whether it has a single or multiple centers. To answer these questions, we evaluated 262 accessions of P. lunatus using two intergenic spacers of chloroplast DNA: atpB-rbcL and trnL-trnF. The data were analyzed using a maximum likelihood tree (ML), a haplotype network and two estimators of genetic differentiation (N _ST and G _ST). Nucleotide diversity (π) and haplotype (Hd) were quantified to estimate the percentage of reduction in genetic diversity (%r) as a founder effect. The ML tree and haplotype network indicated the existence of three groups (AI, MI and MII), which was supported by the high values of N _ST (0.61–0.80). Values for %r were high (58.67–60.83 %). Existence of Mesoamerican and Andean gene pools was confirmed, with two genetically and geographically distinct groups (MI and MII) within the MA gene pool. We present the first evidence for multiple origins of domestication for the MA gene pool. For MI, we propose western central Mexico as the domestication area and between Guatemala and Costa Rica for MII. We observed a founder effect in the MA gene pool as a result of domestication.     

Genetic diversity,population structure and differentiation of rice species from Niger and their potential for rice genetic resources conservation and enhancement

Rice genetic resources conservation and evaluation is crucial to ensure germplasm sources for further crop breeding. We conducted a wide collection of Oryza species in Niger and characterize its diversity with microsatellites (or simple sequence repeats, SSR). The aims of this research were to get a better understanding of the extent of genetic diversity, its structure and partition within rice eco-geographical zones of Niger. There were 264 accessions found in farmers’ and other fields: 173 O. sativa (Asia’s rice), 65 O. glaberrima (Africa’s rice), 25 O. barthii, and 1 O. longistaminata (weedy perennial rice), which were genotyped with 18 SSR. A total of 178 alleles were detected, with a mean of 9.89 alleles per locus. The polymorphism information content was 0.65 and heterozygosity was estimated as 0.14. Two main well-differentiate genotypic groups, which correspond to Asian and African rice species, were identified. The SSR set divided the Asia’s rice group (solely indica) into irrigated and floating rice, with rainfed lowland rice in between. The African rice species group was composed of O. glaberrima, O. longistaminata and O. barthii accessions, but without any clear genetic differentiation among them likely due admixtures within the samples of O. barthii. Five accessions that could be natural interspecific hybrids were too admixed for assigning them to any of the two well-differentiated groups. The partitioning of the overall diversity showed that maximum variation was within genotypic groups and subgroups or cropping ecologies, rather than between eco-geographical zones. The eco-geographical distribution of the diversity suggests germplasm exchange in Niger. Next-steps for conserving rice and crop wild relatives in Niger could be taken using the findings of this research.     

Phylogeographic study based on intraspecific sequence variation of chloroplast DNA for the conservation of genetic diversity in the Japanese endangered species Primula sieboldii

To gather information on intraspecific phylogeography for use in conservation programs for the endangered species Primula sieboldii in Japan, we analyzed sequence variation in five noncoding regions of chloroplast DNA. Twenty-two distinct haplotypes were recognized in total. The distribution of most haplotypes was geographically confined, but one haplotype was widely distributed throughout northern Japan, and several haplotypes were found in geographically distant regions. Three major clades were revealed by phylogenetic analysis of the haplotypes. Clade I was distributed in Kyushu and central Honshu, clade II in western Honshu and Hokkaido, and clade III from central Honshu to Hokkaido. According to analysis of molecular variance, 59.9% of the total cpDNA variation existed among regions, 32.5% among populations within regions, and 7.6% within populations. Therefore, if genetic conservation of the species is valued, transplanting of P. sieboldii among regions should be avoided. Multiple lineages often existed even in geographically narrow areas (e.g., within a 20-km range), so transplantation between adjacent populations in restoration activities should be carefully designed so as not to change the gene pool of local populations significantly. Also, the geographical distribution of cpDNA haplotypes may allow us to confirm the origin of plants collected for commercial purposes.     

Use of Enzymes for the Separation of Protein from Rice Flour

When rice flour was treated with heat stable α-amylases, the effectiveness of protein separation increased with increased temperature. Depending on the enzyme, treatment at 90°C for 45 min resulted in protein contents of 47–65% for the insoluble fraction. Prior gelatinization enhanced the effectiveness of the enzyme reaction but was undesirable because the increased viscosity and gelation could cause difficulties in the processing operation. Follow-up treatment with other carbohydratehydrolyzing enzymes, such as glucoamylase, cellulase, and hemicellulase further increased the protein content up to 76% for the insoluble fraction. The subunit structure of the isolated proteins, based on electrophoretic analysis, remained practically unchanged after the treatment. The limited solubility and emulsion activity of rice protein were also unchanged.     

Use of the 1H nuclear magnetic resonance spectra signals from polyphenols and acids for chemometric characterization of cider apple juices

The low field region (5.8-9.0 ppm) corresponding to aromatic protons and the region 1.8-3.0 ppm of the (1)H NMR spectra were used for characterization and chemometric differentiation of 52 apple juices obtained from six cider apple varieties. The data set consisted of 14 integrated areas corresponding to resonances from acids and phenolic compounds. Multivariate procedures based on hierarchical cluster and discriminant analysis were performed on selected signals of the spectra to determine whether it was possible to distinguish the different juices. Cluster analysis was able to satisfactorily classify the six apple varieties. Discriminant analysis, by means of stepwise procedure for variables selection and leave-one-out for cross-validation, was applied to 40 samples from the year 2001, obtaining recognition and prediction abilities of 100%. The most discriminant variables corresponded to poliphenols, (-)-epicatechin, phloridzin-phloretin, and p-coumaric, chlorogenic, and malic acids. The classification model was applied to 12 samples from apples harvested in the years 2002 and 2003, and the prediction ability was 91.7%.     

Impacts on Streams from the Use of Sulfur-Based Compounds for Dechlorinating Industrial Effluents

We evaluate environmental impacts associated with sulfur-baseddechlorinating agents (sodium bisulfite and sodium thiosulfate)commonly used in industrial wastewater treatment by presentingdata from two examples for Department of Energy facilities inTennessee and Kentucky. One case involved a fish kill (> 24,000 fish) caused directly by sodium bisulfite; the second describes a near-miss situation resulting from over-dosing with sodium thiosulfate. Toxicity tests showed that overfeed situations withsodium thiosulfate or sodium bisulfite can depress pH and dissolved oxygen, causing mortality of fish. Bacteria also can metabolize some sulfur-based dechlorinating agents, thereby increasing the potential for reductions in pH and concentrationsof dissolved oxygen. Although removing toxic levels of chlorine is important when releasing chlorine-containing wastewaters to aquatic systems, waste-treatment plant operators should also beaware of significant impacts that can occur if sulfur-based dechlorinating agents are used to excess.     

Simple and rapid method for the differentiation of Lens culinaris Medik. from false lentil species

The practice of using both common vetch (Vicia sativa L.) and single-flower vetch (Vicia articulata Hornem.) seeds as food or feed is encouraged by the very high resemblance of their seeds with those of small-seeded lentil cultivars. Among the Vicieae, antinutritional and toxic factors are particularly important, because many species, containing high levels of these compounds, are not safe. A simple and fast capillary electrophoresis (CE) method was proposed for the differentiation of lentil cultivars from false lentil species (i.e., single-flour vetch and common vetch). Proteins were extracted from defatted milled seeds with an alcoholic/saline solution. Extracts were separated in an uncoated fused-silica capillary with iminodiacetic acid (IDA) isolectric buffer containing 0.05% hydroxypropylmethylcellulose (HPMC) and 20% acetonitrile. The presented method is useful also for the detection of contamination of whole or split seeds of lentil by vetch species. With respect to alternative techniques, such as DNA-based markers or thin-layer chromatography (TLC), CE has the advantages of being less expensive, faster, and fully automated.     

Use of beer bran as an adsorbent for the removal of organic compounds from wastewater

Beer bran was found to effectively adsorb several organic compounds, such as dichloromethane, chloroform, trichloroethylene, benzene, pretilachlor, and esprocarb. Equilibrium adsorption isotherms conformed to the Freundlich isotherm (log-log linear). Adsorption of these organic compounds by beer bran was observed in the pH range of 1-11. At equilibrium, the adsorption efficiency of beer bran for benzene, chloroform, and dichiloromethane was higher than that of activated carbon. The removal of these organic compounds by beer bran was attributed to the uptake by intracellular particles called spherosomes. The object of this work was to investigate several adsorbents for the effective removal of organic compounds from wastewater.     

Evidence for the Use of Low-Grade Weirs in Drainage Ditches to Improve Nutrient Reductions from Agriculture

Typical controlled drainage structures in drainage ditches provide drainage management strategies for isolated temporal periods. Innovative, low-grade weirs are anticipated to provide hydraulic control on an annual basis, as well as be installed at multiple sites within the drainage ditch for improved spatial biogeochemical transformations. This study provides evidence toward the capacity of low-grade weirs for nutrient reductions, when compared to the typical controlled drainage structure of a slotted riser treatment. Three ditches with weirs were compared against three ditches with slotted risers, and two control ditches for hydraulic residence time (HRT) and nutrient reductions. There were no differences in water volume or HRT between weired and riser systems. Nutrient concentrations significantly decreased from inflow to outflow in both controlled drainage strategies, but there were few statistical differences in N and P concentration reductions between controlled drainage treatments. Similarly, there were significant declines in N and P loads, but no statistical differences in median N and P outflow loads between weir (W) and riser (R) ditches for dissolved inorganic phosphate (W, 92%; R, 94%), total inorganic phosphate (W, 86%; R, 88%), nitrate-N (W, 98%; R, 96%), and ammonium (W, 67%; R, 85%) when nutrients were introduced as runoff events. These results indicate the importance of HRT in improving nutrient reductions. Low-grade weirs should operate as important drainage control structures in reducing nutrient loads to downstream receiving systems if the hydraulic residence time of the system is significantly increased with multiple weirs, as a result of ditch length and slope.     

Synthesis of 1-piperideine-6-carboxylic acid produced by L-lysine-epsilon-aminotransferase from the Streptomyces clavuligerus gene expressed in Escherichia coli

The gene (lat) encoding L-lysine epsilon-aminotransferase (LAT) in Streptomyces clavuligerus was cloned and expressed in Escherichia coli. Nucleotide sequence analysis of lat predicted a single open reading frame (ORF) of 1371 bp, encoding a polypeptide of 457 amino acids with calculated molecular mass of 49.89 kDa. S. clavuligerus LAT was grouped into aminotransferase subfamily II of alpha family on the basis of sequence homology. A model system composed of the recombinant LAT in phosphate buffer was set up to study the biosynthesis of 2-acetyltetrahydropyridine. Lysine was found to be transformed to 1-piperideine-6-carboxylic acid. 2-Acetyltetrahydropyridine was characterized from the mixture of 1-piperideine-6-carboxylic acid and methylglyoxal. For the first time, we demonstrated that the L-lysine epsilon-aminotransferase is responsible for the formation of 1-piperideine-6-carboxylic acid, which may react with methylglyoxal to generate the acylated N-heterocyclic odorant 2-acetyltetrahydropyridine.     

Use of RAPD markers for the study of microbial community similarity from termite mounds and tropical soils

In this study, we test the use of the RAPD (Random Amplified Polymorphic DNA) molecular markers as a way to estimate the similarity of the microbial communities in various termite mounds and soils. In tropical ecosystems, termite activities induce changes in the chemical and physical properties of soil. The question then arises as to whether or not termites affect the presence of natural microbial communities. Successful 16S rDNA amplifications provided evidence of the occurrence of bacterial DNA in termite constructions including both soil feeder and fungus grower materials. A phenetic dendrogram using the similarity distance calculated from pairwise data including 88 polymorphic RAPD markers was reconstructed and bootstrap scores mapped. The microbial communities of the mounds of the four soil-feeding termites were clustered in the same clade, while those of the mounds of the fungus-growing species were distinct like those of control soils. Microbial changes in nests result from termite building behavior, depending on whether they include feces in their constructions for soil-feeders or use saliva as particle cement for fungus-growers. It is argued that RAPDs are useful markers to detect differences in microbial community structure not only between termitaries and control soils but also between mounds of soil-feeders.     

Use of Modified Zeolites for the Remediation of Waters and Cultivated Soils from Cu(II)

The ability of iron oxides (goethite) and natural zeolite (clinoptilolite) to adsorb copper from its aqueous solutions was extensively studied in the past. In this paper, the production of modified zeolites (systems I and II) from raw materials of zeolite and goethite was investigated compared to the initial materials. These two systems presented higher adsorption than goethite or clinoptilolite. Comparing the two systems, system II presented higher adsorption than system I. Isotherm studies showed that the adsorption data from all materials were best described by Freundlich isotherm. According to thermodynamic study, the values of ??G° became more negative with the increase of temperature, indicating that the adsorption process was more favorable to higher temperature. The positive values of ??H° indicated endothermic nature of adsorption. The use of all adsorptive materials as soil improvements was also investigated. Goethite, zeolite, and systems I and II were mixed separately with three different soils (clay loam, sandy loam, and loamy sand). Lettuces were cultivated, and the combination of system II with the loamy sand soil led to the best morphological characteristics of lettuces with the minimum adsorption of copper. Consequently, modified zeolites could be considered as two satisfactory detergents of highly copper concentration in soil and water.     

Use of reverse micelles for the simultaneous extraction of oil, proteins, and glucosinolates from cruciferous oilseeds

Cruciferous oilseeds are important sources of oil, proteins, and glucosinolates (GLs), potentially available when biorefinery processes are used. The proposed extraction technology is based on the use of reverse micelles (RMs) made with cetyltrimethylammonium bromide (CTAB) dispersed in organic solvent. The physicochemical properties of this extraction system and the good water solubility of many high value compounds, such as GLs and some proteins, permit the simultaneous extraction of oil, and these products from cruciferous oilseed meals. This procedure is based on three main steps: (i) seed conditioning; (ii) solid-liquid extraction by RM solution; and (iii) back-transfer of the RM solution for recovery of the extracted compounds. The method makes it possible to simultaneously extract almost the same amount of oil as with pure organic solvents used in the current extraction plants and more than 90% of soluble proteins and GLs. It is a promising biorefinery technology alternative to traditional oil extraction processes.     

Application of multivariate analysis and artificial neural networks for the differentiation of red wines from the Canary Islands according to the island of origin

Eleven metals (K, Na, Ca, Mg, Fe, Cu, Zn, Mn, Sr, Li, and Rb) were determined in 83 red wines from the Canary Islands. The wines presented high concentrations of Na, and the concentrations of Cu and Zn were much lower than the maximum concentrations established by the International Office of Vine and Wine (OIV). Applying principal component analysis, the dimension space was reduced to five principal components that explain 76.4% of the total variance, and the wines tend to separate on the basis of the island of production. Linear discriminant analysis (LDA) allowed a reasonable classification of wines according to the island of production. When artificial neural networks (Kohonen self-organizing maps and back-propagation feed-forward as unsupervised and supervised techniques, respectively) were applied on the matrix of data constituted by the analyzed metals, the results improved in relation to those obtained by other multivariate methods observing a differentiation of wines according to island of production.     

Use of fiber interface direct mass spectrometry for the determination of volatile flavor release from model food systems.

Described in this paper is a fiber interface direct headspace mass spectrometric system for the real-time measurement of flavor release. The system was optimized for the detection of the garlic aroma volatile, diallyl disulfide, from water. Parameters investigated included interface temperature, flow rate through the fiber, flow rate through the sample vessel, and sample stir rate. The delay time for detection of sample after introduction into the sample vessel was determined as 43 s. The system proved to be reliable and robust with no loss in sensitivity or contamination of the mass spectrometer over a 6 month period. The technique was applied to a homologous series of aliphatic alcohols from C(2) to C(7). Results showed that as polarity decreased with increasing chain length the release of volatile into the headspace was faster and gave a higher maximum intensity. Release of the garlic aroma volatile from different commercial mayonnaise products clearly showed a decrease in the release of diallyl disulfide as fat content increased. These results demonstrate the potential of using this technique as a tool for understanding the complex interactions that occur between flavor compounds and the bulk food matrix.