Molecular Basis for Resistance to Fluazifop-P-Butyl in Itchgrass (Rottboellia cochinchinensis) from Costa Rica

Rottboellia cochinchinensis is an annual grass weed species known as itchgrass, or "caminadora" in America´s Spanish speaking countries, and has become a major and troublesome weed in several crops. The application of fluazifop-P-butyl at recommended rates (125 g a.i. ha-1) was observed to be failing to control itchgrass in a field in San José, Upala county, Alajuela province, Costa Rica. Plants from the putative resistant R. cochinchinensis population survived fluazifop-P-butyl when treated with 250 g a.i. ha-1 (2X label rate) at the three- to four-leaf stage under greenhouse conditions. PCR amplification and sequencing of partial carboxyl transferase domain (CT) of the acetyl-CoA carboxylase (ACCase) gene were used to determine the molecular mechanism of resistance. A single non-synonymous point mutation from TGG (susceptible plants) to TGC (putative resistant plants) that leads to a Trp-2027-Cys substitution was found. This Trp-2027-Cys mutation is known to confer resistance to all aryloxyphenoxyproprionate (APP) herbicides to which fluazifop-P-butyl belongs. To the best of our knowledge, this is the first report of fluazifop-P-butyl resistance and a mutation at position 2027 for a Costa Rican R. cochinchinensis population.

Natural abundance of 13C in tropical grasses from the INPA, Instituto Nacional de Pesquisas da Amazônia, herbarium

The stable carbon isotopic composition of 165 grass species was determined with the objective of verifying their photosynthetic pathway (C3 and C4). The samples, taken from the INPA herbarium, were mainly collected in the North of Brazil. Approximately 60% of the species proved to be of the C4 type, with d 13C values ranging from -13.6 to -9.5‰, while the remainder 40% belonged to the C3 type, with values ranging from -34.7 to -23.4‰. This relatively high proportion of C3 species is probably due to the high relative humidity of the sites where the species were collected.

Diurnal variations of non-structural carbohydrates in vegetative tissues of Melinis minutiflora, Echinolaena inflexa and Lolium multiflorum (Poaceae)

Echinolaena inflexa (Poir.) Chase is a native C3 grass species with high biomass production and abundance in the cerrado comparable to Melinis minutiflora Beauv., a C4 forage grass of African origin, widespread in the cerrado, displacing some native herbaceous species. These species accumulate mainly starch, differently of Lolium multiflorum Lam. ssp. italicum Beck cv. Lema (ryegrass), a C3 temperate forage grass that accumulates fructans as the major storage carbohydrate in the vegetative tissues. In the present study, we analyzed diurnal variations in the content and composition of non-structural carbohydrates (NSC) in the aerial vegetative parts of the three species. Results showed similarity in water-soluble carbohydrates (WSC) and starch contents between Melinis minutiflora and Echinolaena inflexa, with the exotic grass accumulating higher amounts of NSC. However, the levels of sucrose, glucose and fructose were 10 fold higher in Lolium multiflorum, that accumulates fructan as the main storage carbohydrate. Although the contents of WSC and starch were similar in the tropical species E. inflexa and M. minutiflora, their diurnal variations were different and could be related to starch metabolism, especially in M. minutiflora.

Hypoglycemic activity of polysaccharide fractions containing ß-glucans from extracts of Rhynchelytrum repens (Willd.) C.E. Hubb., Poaceae

ß-Glucans are soluble fibers with physiological functions, such as interference with absorption of sugars and reduction of serum lipid levels. The objective of the present study was to analyze the distribution of ß-glucans in different tissues of the African grass species Rhynchelytrum repens and also to evaluate their hypoglycemic activity. Leaf blades, sheaths, stems, and young leaves of R. repens were submitted to extraction with 4 M KOH. Analysis of the fractions revealed the presence of arabinose, glucose, xylose, and traces of rhamnose and galactose. The presence of ß-glucan in these fractions was confirmed by hydrolyzing the polymers with endo-ß-glucanase from Bacillus subtilis, followed by HPLC analysis of the characteristic oligosaccharides produced. The 4 M KOH fractions from different tissues were subjected to gel permeation chromatography on Sepharose 4B, with separation of polysaccharides with different degrees of polymerization, the highest molecular mass (above 2000 kDa) being found in young leaves. The molecular mass of the leaf blade polymers was similar (250 kDa) to that of maize coleoptile ß-glucan used for comparison. The 4 M KOH fraction injected into rats with streptozotocin-induced diabetes showed hypoglycemic activity, reducing blood sugar to normal levels for approximately 24 h. This performance was better than that obtained with pure ß-glucan from barley, which decreased blood sugar levels for about 4 h. These results suggest that the activity of ß-glucans from R. repens is responsible for the use of this plant extract as a hypoglycemic drug in folk medicine.

Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae)

Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5% propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1% of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme.

Obtention and characterization of gluten-free baked products

This work had as objective the development of gluten-free breads and muffins using rice flour and maize and cassava starches. From seven samples resulting from a Simplex-Centroid design, the sensory and instrumental analyses of specific volume, elasticity, and firmness were performed. For the sensory analysis, the optimized formulation contained 50% of rice flour and 50% of cassava starch, and for the instrumental evaluation, the optimal simultaneous point for the three conducted analyses were 20% of rice flour, 30% of cassava starch, and 50% of maize starch. A comparative analysis of specific volume, elasticity, firmness, and triangular test was performed with pre-baked, baked, and frozen bread. Physicochemical, nutritional, and microbiological analyses were performed for both bread and muffin according to the Brazilian legislation.

Evaluation of the in vitro glycemic index of a fiber-rich extruded breakfast cereal produced with organic passion fruit fiber and corn flour

The aim of this study was to determine the influence of process parameters and Passion Fruit Fiber (PFF) addition on the Glycemic Index (GI) of an extruded breakfast cereal. A 2³ Central Composite Rotational Design (CCRD) was used, with the following independent variables: raw material moisture content (18-28%), 2nd and 3rd barrel zone temperatures (120-160 ºC), and PFF (0-30%). Raw materials (organic corn flour and organic PFF) were characterized as to their proximate composition, particle size, and in vitro GI. The extrudates were characterized as to their in vitro GI. The Response Surface Methodology (RSM) and Principal Component Analysis (PCA) were used to analyze the results. Corn flour and PFF presented 8.55 and 7.63% protein, 2.61 and 0.60% fat, 0.52 and 6.17% ash, 78.77 and 78.86% carbohydrates (3 and 64% total dietary fiber), respectively. The corn flour particle size distribution was homogeneous, while PFF presented a heterogeneous particle size distribution. Corn flour and PFF presented values of GI of 48 and 45, respectively. When using RSM, no effect of the variables was observed in the GI of the extrudates (average value of 48.41), but PCA showed that the GI tended to be lower when processing at lower temperatures (<128 ºC) and at higher temperatures (>158 ºC). When compared to white bread, the extrudates showed a reduction of the GI of up to 50%, and could be considered an interesting alternative in weight and glycemia control diets.

Characterization of Brazilian wheat cultivars for specific technological applications

Functional and technological properties of wheat depend on its chemical composition, which together with structural and microscopic characteristics, define flour quality. The aim of the present study was to characterize four Brazilian wheat cultivars (BRS Louro, BRS Timbauva, BRS Guamirim and BRS Pardela) and their respective flours in order to indicate specific technological applications. Kernels were analyzed for test weight, thousand kernel weight, hardness, moisture, and water activity. Flours were analyzed for water activity, color, centesimal composition, total dietary fiber, amylose content and identification of high molecular weight glutenins. The rheological properties of the flours were estimated by farinography, extensography, falling number, rapid visco amylography, and glutomatic and glutork equipment. Baking tests and scanning electron microscopy were also performed. The data were subjected to analysis of variance and principal component analysis. BRS Timbauva and BRS Guamirim presented results that did not allow for specific technological application. On the other hand, BRS Louro presented suitable characteristics for the elaboration of products with low dough strength such as cakes, pies and biscuits, while BRS Pardela seemed suitable for bread and pasta products.

Iron enriched Saccharomyces cerevisiae maintains its fermenting power and bakery properties

Iron is an essential micronutrient in the metabolism of almost all living organisms; however, its deficiency is well documented especially in pregnant women and in children. Iron salts as a dietary supplement have low bioavailability and can cause gastrointestinal discomforts. Iron enriched yeasts can provide a supplementation of this micronutrient to the diet because this mineral has a better bioavailability when bonded to yeast cell macromolecules. These yeasts can be used as feed supplement for human and animals and also as baker's yeast. Baker's yeast Saccharomyces cerevisiae was cultivated in a reactor employing yeast media supplemented with 497 mg ferrous sulfate.L-1, and the resultant biomass incorporated 8 mg Fe.g-1 dry matter. This biomass maintained its fermenting power regarding both water displace measurement through carbonic dioxide production and bakery characteristics. The bread produced using the yeast obtained by cultivation in yeast media supplemented with iron presented six times more iron than the bread produced using the yeast obtained by cultivation without iron supplementation.

Effect of storage and processing of Brazilian flaxseed on lipid and lignan contents

Flaxseed has been widely studied around the world; its incorporation into products habitually consumed by human populations has been stimulated due to its unique nutritional value. The objective of this study was to evaluate the chemical composition of Brazilian flaxseed, to analyze the stability of lipids present in whole flaxseed flour (WFF) or partially defatted flaxseed flour (DFF) stored under several temperatures, and to investigate the effect of bread making on a product containing flaxseed. Whole flaxseed flour presented (g.100 g-1) 25.7 of insoluble fiber, 10.7 of soluble fiber, 38.9 of lipids, and 2.65 of lignan. Defatted flaxseed flour presented 65% less lipids, 36% more fiber and 56% more lignan than whole flaxseed flour. The fatty acid profile was maintained in the defatted flaxseed flour, and it presented a stable composition during storage under ambient temperature, refrigeration, and freezing. The fatty acid profile was similar in the bread containing defatted flaxseed flour after dough development, baking, and storage at room temperature or refrigerated. After baking, 89% of the lignan content was kept in bread. Results show that Brazilian flaxseed has an interesting chemical composition, and that defatted flaxseed, by-product of lipid extraction, presents a good stability to grind and storage under several temperatures. Thus, defatted flaxseed flour can be incorporated in bread, increasing its nutritional and functional value.

Characterization and effect of clarified araçá (Psidium guineenses Sw.) juice on postprandial glycemia in healthy subjects

Brazilian native fruits are excellent sources of bioactive compounds of phenolic nature. Some of these compounds are able to inhibit carbohydrate- metabolizing enzymes (in vitro), α-amylase and α-glucosidase, delaying carbohydrate digestion. This study aimed to evaluate the effect of clarified araçá (Psidium guineenses Sw.) juice on postprandial glycemia in humans after consumption of 25 g of available carbohydrates (approximately 50 g of white bread) and characterize the phenolic compounds and in vitro antioxidant capacity of araçá juice and pulp. The results showed that the clarified juice had a positive effect on postprandial glycemia reducing the total amount of glucose absorbed, lengthening the time to reach maximum blood glucose concentration, reducing glucose incremental velocity, and decreasing glucose incremental percentage. Both frozen pulp and clarified juice had high amounts of phenolic compounds, antioxidant capacity, and proanthocyanidins, among which oligomers (monomers to tetramers), pentamers, hexamers, heptamers, octamers, nonamers, decamers, and polymers were detected, and they are probably associated with in vivo effects.

Effect of high and low molecular weight glutenin subunits, and subunits of gliadin on physicochemical parameters of different wheat genotypes

Identification of functional properties of wheat flour by specific tests allows genotypes with appropriate characteristics to be selected for specific industrial uses. The objective of wheat breeding programs is to improve the quality of germplasm bank in order to be able to develop wheat with suitable gluten strength and extensibility for bread making. The aim of this study was to evaluate 16 wheat genotypes by correlating both glutenin subunits of high and low molecular weight and gliadin subunits with the physicochemical characteristics of the grain. Protein content, sedimentation volume, sedimentation index, and falling number values were analyzed after the grains were milled. Hectoliter weight and mass of 1000 seeds were also determined. The glutenin and gliadin subunits were separated using polyacrylamide gel in the presence of sodium dodecyl sulfate. The data were evaluated using variance analysis, Pearson's correlation, principal component analysis, and cluster analysis. The IPR 85, IPR Catuara TM, T 091015, and T 091069 genotypes stood out from the others, which indicate their possibly superior grain quality with higher sedimentation volume, higher sedimentation index, and higher mass of 1000 seeds; these genotypes possessed the subunits 1 (Glu-A1), 5 + 10 (Glu-D1), c (Glu-A3), and b (Glu-B3), with exception of T 091069 genotype that possessed the g allele instead of b in the Glu-B3.

Non-parametric stability analyses of dough properties in wheat

AbstractLiterature has unveiled that a paper has not been published yet on using non-parametric stability statistics (NPSSs) for evaluating genotypic stability in dough properties of wheat. Accordingly, the effects of genotype (G), environment (E) and GE interaction (GEI) on alveograph parameters, i.e. dough baking strength (W) and its tenacity (P)/extensibility (L), of 18 wheat (T. aestivum L.) genotypes were studied under irrigated field conditions in an 8-year trial (2006-2014) in central Turkey. Furthermore, genotypic stability for W and P/L was determined using 8 NPSSs viz. RM-Rank mean, RSD-Rank’s standard deviation, RS-Rank Sum, TOP-Ranking, Si(1), Si(2), Si(3) and Si(6) rank statistics. The ANOVA revealed that W and P/L were primarily controlled by E, although G and GEI also had significant effects. Among the 8 NPSSs, only RM, RS and TOP statistics were suitable for detecting the genotypes with high stable and bread making quality (e.g. G1 and G17). In conclusion, using RM, RS and TOP statistics is advisable to select for dough quality in wheat under multi-environment trials (METs).

Sulphuric acid scarification effects on Brachiaria brizantha, B. humidicola and Panicum maximum seed dormancy release

Most tropical forage grass species have dormant seeds, which reduce percentages in germination tests. The objective of this study was to evaluate H2SO4 scarification effects on seed dormancy releasing, through germination time (T50) and variability among germination test replicates, in 630, 94 and 82 seed samples of B. brizantha, B. humidicola and P. maximum, respectively, tested at the Central Seed Testing Laboratory, Campinas, Brazil, from 1991 to 1999. Germination tests used two 4 x 100 replicates of intact and scarified seeds (15-, 10-, 5-minute treatments, respectively). Mean germination time (T50) and variability among germination replicates were also analysed. Statistical analysis was performed by t-test paired samples for means. Scarification promoted general decreases in T50, while variability among germination test replicates was reduced in B. brizantha. Scarification increase germination in B. brizantha and P. maximum, but is deleterious in B. humidicola.