Corn Endosperm Fermentation Using Endogenous Amino Nitrogen Generated by a Fungal Protease

Fractionating the corn kernel to separate endosperm from germ and pericarp improves corn ethanol processing by increasing fermentation throughput and generating salable coproducts. One fractionation technology, dry fractionation (DF), suffers from loss of germ‐derived nutrients and amino acids, resulting in poor fermentation performance. Such deficiencies may be addressed by increasing nitrogen and other nutritional supplementation. As an alternative to exogenous nitrogen source, we investigated the use of a fungal protease to generate free amino nitrogen (FAN) from corn endosperm. Incubation of endosperm with protease did not affect subsequent liquefaction and saccharification. FAN supplementation through proteolysis resulted in fermentation being 99% complete in 48 hr, compared to 93% maximum with urea supplementation. Viable cell growth rates were similar in FAN and urea‐supplemented fermentations. Urea and FAN addition resulted in similar fermentation characteristics and similar FAN consumption rates as with FAN alone, which was indicative that FAN was assimilated preferentially. Increased amounts of maltose remaining after fermentation were correlated with initial FAN concentrations in mash. This observed trend was implicated in ethanol yield reduction of 2 g/L at high protease loading (generating 1.6 mg of FAN/g of glucose substrate) compared to a urea control. Using a glucose and maltose solution, we confirmed higher residual maltose in fermentations supplemented with high FAN concentrations. Use of protease to generate optimal FAN concentration in mash (1.2 mg of FAN/g of glucose substrate) could improve economics of dry fractionated corn ethanol production by increasing fermentation rates and, consequently, reducing fermentation time.     

Physicochemical properties of native and recombinant mungbean (Vigna radiata L. Wilczek) 8S globulins and the effects of the N-linked glycans

We have previously cloned and characterized the cDNAs of three isoforms of the 8S globulin of mungbean, expressed the major 8Salpha isoform in Escherichia coli, and purified and successfully crystallized it (Bernardo, A. E. N.; Garcia, R. N.; Adachi, M.; Angeles, J. G. C.; Kaga, A; Ishimoto, M.; Utsumi, S.; Tecson-Mendoza, E. M. J. Agric. Food Chem. 2004, 52, 2552-2560). Herein, we report the physicochemical and emulsifying properties of the native 8S and recombinant 8Salpha globulin or vicilin. The circular dichroism spectra analysis of the native 8S and recombinant 8Salpha globulins revealed that the recombinant 8Salpha formed a secondary structure close to that of the native 8S. Further, gel filtration analysis showed that 8Salpha was able to assemble into trimers. The native 8S and recombinant 8Salpha globulins were soluble at pH 3.4 and at pH 7.4-9.0 at low ionic strength, mu = 0.08. Interestingly, the native 8S was more soluble at pH 7.0 and pH 7.4 than the recombinant 8Salpha at mu = 0.08. Both forms were very soluble at pH 3.4-9.0 at high ionic strength, mu = 0.50. The native form exhibited a higher T(m) (69.2, 79.5, and 83.8 degrees C) than the recombinant form (65.6, 71.6, 77.5 degrees C) at mu = 0.1, 0.2, and 0.5, respectively. The recombinant form was found to have greater surface hydrophobicity than the native form. There was little difference in the emulsifying ability between the native 8S and 8Salpha at pH 3.4 and pH 7.6. The results indicate that the presence of N-linked glycans is not essential in the assembly and stable conformation of the mungbean vicilin. However, the N-linked glycans might have contributed to the higher solubility at low ionic strength, greater thermal stability, and decreased surface hydrophobicity of the native vicilin as compared to the recombinant 8Salpha. On the other hand, the N-linked glycans showed little effect on the emulsifying ability of the protein.     

Topography and crop management are key factors for the development of american leaf spot epidemics on coffee in costa rica

ABSTRACT We monitored the development of American leaf spot of coffee, a disease caused by the gemmiferous fungus Mycena citricolor, in 57 plots in Costa Rica for 1 or 2 years in order to gain a clearer understanding of conditions conducive to the disease and improve its control. During the investigation, characteristics of the coffee trees, crop management, and the environment were recorded. For the analyses, we used partial least-squares regression via the spline functions (PLSS), which is a nonlinear extension to partial least-squares regression (PLS). The fungus developed well in areas located between approximately 1,100 and 1,550 m above sea level. Slopes were conducive to its development, but eastern-facing slopes were less affected than the others, probably because they were more exposed to sunlight, especially in the rainy season. The distance between planting rows, the shade percentage, coffee tree height, the type of shade, and the pruning system explained disease intensity due to their effects on coffee tree shading and, possibly, on the humidity conditions in the plot. Forest trees and fruit trees intercropped with coffee provided particularly propitious conditions. Apparently, fertilization was unfavorable for the disease, probably due to dilution phenomena associated with faster coffee tree growth. Finally, series of wet spells interspersed with dry spells, which were frequent in the middle of the rainy season, were critical for the disease, probably because they affected the production and release of gemmae and their viability. These results could be used to draw up a map of epidemic risks taking topographical factors into account. To reduce those risks and improve chemical control, our results suggested that farmers should space planting rows further apart, maintain light shading in the plantation, and prune their coffee trees.     

Effect of NaCl salinity in the genotypic variation of cowpea (Vigna unguiculata) during early vegetative growth

Twenty-five genotypes of cowpea (Vigna unguiculata L. Walp.) were tested for salt-tolerance at the vegetative growth stage in pot in the greenhouse experiments at salinity levels of 0, 85, and 170 mM NaCl. Plant survival was the main criterion for classifying genotypes. Other criteria included the ion concentration (Na⁺ and Cl⁻) in root and shoot and biomass accumulation. Four local accessions (‘Paceño’, ‘Tardón’, ‘Sonorense’, and ‘Cuarenteño’), three accessions from California (‘CB46’, ‘CB27’, and ‘CB3’), and one accession from the International Institute of Tropical Agriculture (IITA) (‘IT82D-889’) survived at concentrations of both 85 and 170 mM NaCl and were classified as salt-tolerant, while ‘IT96D-666’, ‘IT89KD-288’, and ‘IT93K-734’ from IITA were classified as salt-sensitive. One local accession (‘Sesenteño’), three accessions from IITA (‘PEPH-V Wes-85’, ‘IT86D-719’, and ‘IT95K-1090-12’), and one accession from California (‘CB5’) were classified as moderately salt-tolerant. Eight accessions from IITA (‘IT96D-733’, ‘IT90K-277-2’, ‘IT91K-93-10’, ‘IT91K-118-20’, ‘IT90K-284-2’, ‘IT95K-1088-4’, ‘IT89KD-391’, and ‘IT94K-437-1’) and one from California (‘CB88’) were classified as moderately salt-sensitive. Biomass was affected by both 85 and 170 mM NaCl in all groups of genotypes, however, salt-tolerant and moderately salt-tolerant genotypes showed higher biomass than genotypes classified as moderately salt-sensitive and salt-sensitive. In all genotypes Cl⁻ concentration was higher in shoots than roots and increased as salinity increased. Similarly Na⁺ concentration increased with increasing salinity. However, in salt-tolerant and moderately salt-tolerant genotypes, Na⁺ concentration was more in roots than shoots, while in moderately salt-sensitive and salt-sensitive genotypes, Na⁺ was higher in shoots than roots.     

Response of high mountain landscape to topographic variables: Central pyrenees

An objective method for inductively modelling the distribution of mountain land units using GIS managed topographic variables is presented. The landscape of a small high mountain catchment in the Spanish Pyrenees, covered with grassland, was classified in ten land units by hierarchical agglomerative clustering, using a sample of 194 random plots, in which classes of vegetation, soils and landforms were defined. Additionally, seven layers of topographic variables (altitude, slope angle, aspect, solar radiation, topographic wetness index, specific catchment area, and regolith thickness) were created from a Digital Elevation Model. The affinity of each land unit to the topographic variables was calculated using Binary Discriminant Analysis (BDA), after dichotomising the latter around their mean values. Then, the distribution of each land unit was predicted by boolean operations combining step by step distributions for the seven topographic variables ordered, for each unit, after the absolute values of the Haberman’s residuals in BDA. The predicted distributions were tested (χ²) against that of the observed sampling plots. From the original ten land units, the distributions of eight of them were successfully predicted (four are related to the slope sequence, two reflect the water accumulation in the soil, and two respond to geomorphic processes) while the remaining two had to be rejected. Part of the catchment (39%) was not assigned to any land unit, probably because more distributed variables accounting for snow distribution are necessary.     

Growth and nutritional disorders of coffee cultivated in nutrient solutions with suppressed macronutrients

The objective was to evaluate the effect of omitting macronutrients in the nutrients solution on growth characteristics and nutritional status of coffee. The treatments were complete nutrients solutions and solutions with nutrient omission: N (nitrogen), P (phosphorus), K (potassium), Ca (calcium), Mg (magnesium) and S (sulfur). The experiment was carried out under greenhouse conditions with 3 replicates in a completely random design. Plant height, number of leaves per plant, stem diameter, relative chlorophyll index, photosynthesis rate, stomatal conductance, transpiration, carbon dioxide (CO₂) concentration, dry matter, content levels of macronutrients in plant aerial part and root system, and nutritional disorders were evaluated. Macronutrients suppression affected nutrients concentration in many plant parts, inducing the appearance of symptoms characteristic of each nutrient. The most limiting nutrients for coffee plants development were nitrogen and calcium, reflected in the lower dry matter accumulation and nitrogen the most required.     

Macronutrient omission and the development and nutritional status of basil in nutritive solution

The correct nutrition of basil (Ocinum basilicum L.) is important to increase its production and quality; however, few papers have deal with this subject. The aim of this work was to evaluate the effects of omission of individual macronutrients on the growth and nutritional status of basil cultivated in nutritive solution. The treatments consisted of nutrient solutions with nitrogen, phosphorus, potassium, calcium, magnesium or sulfur (N, P, K, Ca, Mg, or S) omissions and a complete solution treatment. The plants were cultivated in 8 L plant pots. Plant height, number of leaves per plant, leaf area, relative chlorophyll index, net photosynthesis rate, stomatal conductance, plant dry matter, concentration levels of macronutrients in the aerial part and root system, and nutritional disorders were all evaluated. Nutrient omission was a limiting factor for plant development, substantially reducing its growth. There was also a considerable decrease in nutrient accumulation when compared to the control treatment.     

Using detection dogs and genetic analyses of scat to expand knowledge and assist felid conservation in Misiones,Argentina

Many carnivores require large ranges to meet their ecological and energetic needs; however, anthropogenic changes threaten species and their habitats. Camera traps have been used to effectively collect data on carnivores in a variety of habitat types; however, a single survey effort is typically limited to species that have similar body size, habitat use and movement patterns, and individual identification of animals is not always possible. We evaluated whether scat detection dogs could effectively survey for 4 wide‐ranging felids that vary in these characteristics: jaguars (Panthera onca), pumas (Puma concolor), ocelots (Leopardus pardalis) and oncillas (Leopardus tigrinus). From June to October 2009 and May to August 2011, a detection dog‐handler team detected 588 scats, from which 176 unique genotypes were detected. We assigned sex to 84.7% of the genotyped scats and identified 55 individuals multiple times. The effectiveness of these noninvasive techniques (detection dogs and genetic analyses of scat) not only opens the door for additional studies in areas that were previously difficult or impossible with standard survey techniques, but also provides conservationists with a set of tools that overcome some of the limitations associated with the use of camera traps alone.     

Effects of simple and double grafting melon plants on mineral absorption,photosynthesis, biomass and yield

The Spanish type cultivar ‘Piel de Sapo’ (Cucumis melo L. var. saccharinus), has a limited compatibility with the Cucurbita maxima × Cucurbita moschata hybrids currently used as rootstocks. Double grafting can be used to improve compatibility between rootstock and scion by means of an intermediate rootstock compatible with both. Non-grafted, single, and double grafted melon plants of the cultivar Piel de Sapo were evaluated for water, nutrient absorption, photosynthesis activity, biomass production in early phases, as well as for yields and fruit quality in a long term trial. The hybrid ‘Shintoza’ (C. maxima × C. moschata) was used as rootstock, and the cantaloupe type melon cultivar ‘Sienne’ as an intermediate scion. Grafting did not affect net photosynthetic values, yet increased water use efficiency (35%). Double grafted plants increased aerial dry weights (66% and 31% with respect to non-grafted and simple graft plants, respectively), and also increased capacity for uptaking beneficial minerals (between 61% and 13% and particularly for NO₃⁻, P, K, Ca, Mn, and Zn) with respect to non-grafted and single grafted plants. The quantum efficiency PSII photochemistry values increased in double grafted plants (12%) with respect to the control plants. Consequently, double grafting on a vigorous rootstock such as ‘Shintoza’, with an intermediate scion, results in improved mineral and water absorption and achieves an increase in ion influx to the scion – so enabling an increase in light photosynthetic reaction and biomass. Double grafted plants increased fruit yield when compared to simple grafted and non-grafted plants (12% and 56%, respectively) and did not affect fruit quality in terms of °Brix and colour. In conclusion, double grafting presents several beneficial aspects that are counter-balanced by the extra cost of the technique. The difference in yields reflects compatibility problems.