Characterization of AFN1, a Gene Associated with Cereal Grain Germination

The AFN1 gene is transiently expressed in germinating oat grains. As AFN1 is not expressed in dormant oat grains during imbibition, we hypothesize that AFN1 may be involved in stimulating the germination process. Sequence analysis of an AFN1 cDNA clone indicates that the AFN1 polypeptide is similar to a previously identified abscisic acid (ABA) glucosyl transferase. This suggests that AFN1 may be acting to glucosylate ABA, thereby inactivating it. As the hormone ABA is known to inhibit germination, ABA glucosylation/inactivation could lead to germination in grains expressing AFN1. To test this hypothesis, we have constructed an expression plasmid that encodes an MBP::AFN1 (maltose binding protein) fusion protein. E. coli cells carrying the expression plasmid were found to produce the MBP::AFN1 fusion protein as a substantial fraction of total protein. We are currently in the process of purifying the MBP::AFN1 fusion protein by affinity chromatography, so that it can be assayed for ABA glucosyl transferase activity. We also wish to test the effect of AFN1 gene expression during grain imbibition on the germination behavior of the grains. To this end, we have constructed plasmids for the overexpression and RNAi-based suppression of AFN1 in transgenic plants. These plasmids have been introduced into oat cells by particle bombardment and we are in the process of regenerating transgenic plants for study.

Comparison Of Demand Pattern Calibration In Water Distribution Networks With Geographic And Non-Geographic Parameterization

Demands are one of the most uncertain parameters in a water distribution network model. A good calibration of the model demands leads to better solutions when using the model for any purpose. A demand pattern calibration methodology that uses a priori information has been developed for calibrating the behaviour of demand groups. Generally, the behaviours of demands in cities are mixed all over the network, contrary to smaller villages where demands are clearly sectorised in residential neighbourhoods, commercial zones and industrial sectors. Demand pattern calibration has a final use for leakage detection and isolation. Detecting a leakage in a pattern that covers nodes spread all over the network makes the isolation unfeasible. Besides, demands in the same zone may be more similar due to the common pressure of the area rather than for the type of contract. For this reason, the demand pattern calibration methodology is applied to a real network with synthetic non-geographic demands for calibrating geographic demand patterns. The results are compared with a previous work where the calibrated patterns were also non-geographic.

Competition of sicklepod, cassia obtusifolia l., densities on soybean, glycine max (l.) merr., at variable row distance

Competition studies with soybeans, Glycine max (L.) Merr. "Bragg." and sicklepod, Cassia obtusifolia L., were conducted at the Agricultural Research and Education Center of the University of Florida in Quincy, Florida. Two field experiments were established, one on May 22, 1975. and the other four weeks later, on June 19, 1975, to determine the competitive effects of various sicklepod densities and the influences of soybean row distances on weed dry matter, soybear plant characteristics, yield components and seed yield, and on soil nutrient content. Control, low, medium, and high sicklepod densities in the first experiment were O, 25,000, 53,000, and 77,000 p1ants/ha, respectively; while the second experiment presented control, low, medium, and high sicklepod densities of O, 36,000, 68,000, and 122,000 plants/ha, respectively. Three soybean row distance treatments were tested using a constant pattern of 90-, 60-, and 45-cm widths throughout the growing season. Three other treatments, evaluated in a variable patern, were initially seeded in 30-cm row widths. Five weeks after planting, an appropriate number of soybean rows were harvested from the 30"cm pattern to establish row distances of 90, 60, and 30-60 cm for the remainder of the season. ln the greenhouse a test was conducted to evaluate the effects af those variables on seed germination and seedling vigor for the next soybean generation. As a result of full-season sicklepod competition, soybean plants were less branched, set fewer leaves, and presented thinner stems as compared to the control. However, height of soybean plants was not affected by the presence of sicklepod. ln one of the two experiments, number of nodes decreased for soybeans under weed campetition. The yield components--number of pods; number of seeds, and seed yield per soybean plant--were all similarly reduced due to weed competition. Seeds per pod were decreased to a lesser extent. Soybean seed yields per unit area were significantly diminished by increasing levels af sicklepod ínfestation. While the control produced 3120 kg/ha, the sicklepod densities of 25,000, 53,000, and 77,000 plants/ha reduced seed yíelds 47, 65, and 73%, respectively. As soybean row distances decreased, number of branches, number of leaves, and stem diameter of soybeans decreased. However, the height of soybean plants increased with narrwing of row width. The components of seed yield--number of pods, number of seeds, and seed yield per soybean plant--diminished as row spacing was reduced. Maximum difference between row distances for these attributes was attained for soybean plants under weed-free conditions. Generally, as row width decreased, soybean seed yield per unit area increased. Specifically, soybear.s in 90-cm rows, either in constant or variable row pattern, yielded less than soybeans in 60- and 30-60-cm rows in the variable pattern. Soil contents of phosphorus, potassium, calcium, and magnesium were not affected by the various levels of sicklepod and soybean populalions. Neither the sicklepod densities nor the soybean row distances influenced seed germination and seedling vigor in the next soybean generation. Sicklepod was a strong competitor with soybeans at all density ranges investigated. Because sicklepod grows taller than soybeans during the reproductive stages of the crop, limited success can be reached by varying row spacing alone. However, this practice is considered an integral measure to complement other methods of sicklepod control. Compared to constant rows, the soybean cropping system using variable row spacings presents the choice of planting soybeans at close row spacings to provide early competition with weeds and the possibility of obtaining a forage crop after the first month of growth, without any decreases on the final seed yields.

Headspace solid-phase microextraction combined with mass spectrometry as a powerful analytical tool for profiling the terpenoid metabolomic pattern of hop-essential oil derived from Saaz variety

Hop(HumuluslupulusL.,Cannabaceaefamily)isprizedforitsessentialoilcontents,usedin beer production and, more recently, in biological and pharmacological applications. In this work,a methodinvolvingheadspace solid-phase microextractionand gas chromatography– mass spectrometry was developed and optimized to establish the terpenoid (monoterpenes and sesquiterpenes) metabolomic pattern of hop-essential oil derived from Saaz variety as a mean to explore this matrix as a powerful biological source for newer, more selective, biodegradable and naturally produced antimicrobial and antioxidant compounds. Different parameters affecting terpenoid metabolites extraction by headspace solid-phase microextraction were considered and optimized: type of fiber coatings, extraction temperature, extraction time, ionic strength, and sample agitation. In the optimized method, analytes were extracted for 30 min at 40 C in the sample headspace with a 50/30 m divinylbenzene/carboxen/polydimethylsiloxane coating fiber. The methodology allowed the identification of a total of 27 terpenoid metabolites, representing 92.5% of the total Saaz hop-essential oil volatile terpenoid composition. The headspace composition was dominated by monoterpenes (56.1%, 13 compounds), sesquiterpenes (34.9%, 10), oxygenated monoterpenes (1.41%, 3), and hemiterpenes (0.04%, 1) some of which can probably contribute to the hop of Saaz variety aroma. Mass spectrometry analysis revealed that the main metabolites are the monoterpene -myrcene (53.0±1.1% of the total volatile fraction), and the cyclic sesquiterpenes, -humulene (16.6 ± 0.8%), and -caryophyllene (14.7 ± 0.4%), which together represent about 80% of the total volatile fraction from the hop-essential oil. Thesefindingssuggestthatthismatrixcanbeexploredasapowerfulbiosourceofterpenoid metabolites.

Solid phase microextraction as a reliable alternative to conventional extraction techniques to evaluate the pattern of hydrolytically released components in Vitis vinifera L. grapes

In present research, headspace solid-phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC–qMS), was evaluated as a reliable and improved alternative to the commonly used liquid–liquid extraction (LLE) technique for the establishment of the pattern of hydrolytically released components of 7 Vitis vinifera L. grape varieties, commonly used to produce the world-famous Madeira wine. Since there is no data available on their glycosidic fractions, at a first step, two hydrolyse procedures, acid and enzymatic, were carried out using Boal grapes as matrix. Several parameters susceptible of influencing the hydrolytic process were studied. The best results, expressed as GC peak area, number of identified components and reproducibility, were obtained using ProZym M with b-glucosidase activity at 35 °C for 42 h. For the extraction of hydrolytically released components, HS-SPME technique was evaluated as a reliable and improved alternative to the conventional extraction technique, LLE (ethyl acetate). HS-SPME using DVB/CAR/PDMS as coating fiber displayed an extraction capacity two fold higher than LLE (ethyl acetate). The hydrolyzed fraction was mainly characterized by the occurrence of aliphatic and aromatic alcohols, followed by acids, esters, carbonyl compounds, terpenoids, and volatile phenols. Concerning to terpenoids its contribution to the total hydrolyzed fraction is highest for Malvasia Cândida (23%) and Malvasia Roxa (13%), and their presence according previous studies, even at low concentration, is important from a sensorial point of view (can impart floral notes to the wines), due to their low odor threshold (μg/L). According to the obtained data by principal component analysis (PCA), the sensorial properties of Madeira wines produced by Malvasia Cândida and Malvasia Roxa could be improved by hydrolysis procedure, since their hydrolyzed fraction is mainly characterized by terpenoids (e.g. linalool, geraniol) which are responsible for floral notes. Bual and Sercial grapes are characterized by aromatic alcohols (e.g. benzyl alcohol, 2-phenylethyl alcohol), so an improvement in sensorial characteristics (citrus, sweet and floral odors) of the corresponding wines, as result of hydrolytic process, is expected.

Influência do sistema anteoxidante da catalase no ciclo do glioxilato durante o estabelecimento pós-germinativo de cártamo (Carthamus tinctorius L.) e de girassol (Helianthus annuus L.)

Oilseeds are a high-value natural resource, due to its use as a substitute for petroleum. However, the storage time can reduce seed viability and oil quality. Therefore, scientific efforts have been made to provide a increment of storage time, germination rates and plant establishment of high-value oilseeds. The seedling establishment depends of the plant pass over the functional transition stage, characterized by a metabolic change from heterotrophic condition to autotrophic one. The storage oil mobilization is performed by β-oxidation process and the glyoxylate cycle. Also, the functional transition involves acclimation to photosynthetic condition, which generally includes the participation of antioxidant system and the reactive oxygen species, the latter are produced in various reactions of primary and secondary metabolism. In the present study, Catalase was inhibited during the functional transition of sunflower and safflower, after were performed many analyzes to elucidate the effects caused on the SOD and APX antioxidant systems. Also, were checked the changes in expression pattern of the glyoxylate cycle enzymes markers, ICL and MLS. It was observed that after CAT inhibition, the SOD and APX antioxidant systems allow the seedling establishment. Besides, was verified that both oilseeds can be accelerate the reverse mobilization and the photosynthetic establishment when Catalase activity has dramatically decreased

Longitudinal distribution and lateral pattern of megalopal settlement and juvenile recruitment of Carcinus maenas (L.) (Brachyura, Portunidae) in the Mira River Estuary, Portugal

Settlement is a critical process in the life history of crabs, and thus affecting the abundance, distribution and structure of estuarine communities. The spatial pattern of settlement of megalopae of the shore crab Carcinus maenas along a longitudinal estuarine gradient (Mira River Estuary, Portugal) was examined, as well as its effects on the juvenile population. To measure megalopal settlement, four replicate collectors were deployed in six equally spaced stations along the estuarine axis. Juveniles were collected on the same locations with a quadrat randomly deployed on the substrate. To assess fine-scale megalopal settlement within a curved region of the estuary, replicate collectors were deployed on both margins along Moinho da Asneira curve. Megalopae settled differently along the six longitudinal points, with a tendency to attenuate their settlement upstream. Within the curved region, megalopae preferentially settled on the left margin collectors, probably due to the weaker velocity speeds felt on this margin. Concerning the overall juvenile density, there were significant differences among the stations distributed along the estuary, but they did no reflect a longitudinal dispersion attenuation pattern. Size-frequency distribution of the juvenile population showed that the average size is higher on the left margin. Recruits (carapace length between 1.0 mm and 3.4 mm) were more abundant on the upstream stations. Density of early juveniles (3.4 mm-6.5 mm) and juveniles (6.5 mm-10 mm) was more stable throughout the estuary axis than that of recruits. This distribution pattern may result from tidal excursion processes or mechanisms to avoid biotic interactions, such as predation and competition. (c) 2006 Elsevier Ltd. All rights reserved.