Evaluation of Chilo partellus and Busseola fusca susceptibility to δ-endotoxins in Bt maize

Susceptibility of Chilo partellus (Lepidoptera, Crambidae) and Busseola fusca (Lepidoptera, Noctuidae) populations to Cry proteins from the bacterium, Bacillus thuringiensis (Bt), the δ-endotoxins Cry1Ab and Cry1Ba in Bt-maize, were evaluated under biosafety greenhouse conditions. Larval feeding on Bt-maize was adjusted to deliver sub-lethal doses of δ-endotoxins from the two events; survivors were reared on artificial diet to obtain successive generations. Eight generations of three C. partellus populations and five generations of a B. fusca population were screened for susceptibility on each event. Mean proportion of surviving larvae from Bt-maize plants, and the corresponding pupal weights of survivors for each population, were lower for individuals exposed to δ-endotoxins. Both Bt Cry proteins expressed in maize leaves controlled C. partellus and showed stability in control, with no indication of a change in susceptibility among generations. Neither toxin, however, provided complete control of B. fusca, but no changes in susceptibility were observed after five generations of selection. Implications for development of future transgenic Bt maize events, and research for East Africa are discussed.     

Characterisation of a ω-gliadin gene in Triticum tauschii

A ω-gliadin gene at the Gli-D^t1 locus of Triticum tauschii accession AUS18913 was isolated using PCR primers, designed from published sequences of ω-gliadin genes of bread wheat cv Cheyenne, and deduced sequences of the N-terminal amino acids of ω-gliadin proteins. Further, the derived protein was isolated from A-PAGE and was sequenced. The protein sequence contained a signal peptide of 19 amino acids followed by a short N-terminal sequence of 11 amino acids, a central repetitive domain that covers approximately 90% of the sequence and a short C-terminal domain of 12 amino acids. The sequence comparison with other ω-gliadins showed a high level of similarities between them. Further analysis of the ω-gliadins using A-PAGE revealed that there are three ω-gliadin proteins in AUS18913 accession. Comparison of N-terminal sequences of these proteins revealed that two of these proteins have very high homologies with ω-gliadins of Cheyenne while the third one was significantly different.     

Identification and characterization by PCR–RFLP analysis of the genetic variation for the Glu-A1x and Glu-B1x genes in rivet wheat (Triticum turgidum L. ssp. turgidum)

Many studies have suggested that the true variation of high-molecular-weight (HMW) glutenin subunits in wheat could be under-estimated due to the anomalous mobility of these proteins in one-dimensional electrophoresis (SDS-PAGE). An alternative technique used is the specific PCR amplification of these genes, however the scarce variation in size among the different alleles could also under-estimate the true variability. In this study, the variability of HMW glutenin subunits found in one Spanish collection of rivet wheat (Triticum turgidum L. ssp. turgidum) was evaluated by a combination of two techniques (PCR amplification and digestion with endonucleases). The data allowed detection of allelic variations that were not clearly detected by SDS-PAGE or PCR analysis alone. The approach used in the current study could allow identification of alleles contributing to achieve desirable quality in modern wheat.     

Are insensitivities of Venturia inaequalis to myclobutanil and fenbuconazole correlated?

In the UK, two demethylation inhibitor (DMI) fungicides, myclobutanil and fenbuconazole, are recommended for controlling apple scab. Experiments were conducted to determine whether the sensitivity of V. inaequalis to myclobutanil was correlated with that to fenbuconazole. There was a marked reduction in the sensitivity of V. inaequalis to myclobutanil; averaged ED₅₀ values increased from 0.338 mg L⁻¹ in the baseline population to 2.945 mg L⁻¹ for isolates from a commercial orchard. Overall, the average ED₅₀ value for fenbuconazole was only about 20% of that for myclobutanil for the baseline population. There was an overall significant positive correlation in the fungal sensitivities to myclobutanil and fenbuconazole, but such correlation only accounted for 40% of the total observed variability. In addition, the magnitude of this cross-resistance varied among orchards. The finding is discussed in the context of fungicide deposition in relation to disease control.     

Pasting characteristics and in vitro digestibility of γ-irradiated RS4 waxy maize starches

The effect of γ-irradiation on the physicochemical properties of cross-linked waxy maize resistant starches was examined. The cross-linked waxy maize starches contained resistant starch (RS) of 56.1 and 63.5%, respectively for 5 and 10% sodium trimetaphosphate (STMP)/sodium tripolyphosphate (STPP) cross-linking, and the RS contents slightly decreased as the irradiation dose increased whereas the RS content in unmodified waxy maize starch increased with an increase in irradiation dose. For both native and cross-linked starches, the rapidly digestible starch (RDS) content increased and the slowly digestible starch (SDS) content decreased by the irradiation. The solubility of the native and cross-linked starches increased as the irradiation dose increased. The cross-linked starches did not swell in boiling water without showing pasting viscosity. However, the starches became swellable, forming pastes by irradiation, and the pasting viscosity gradually increased with an increase in irradiation dose. The crystallinity as determined by an X-ray diffraction analysis remained unchanged upon cross-linking and γ-irradiation. However, the gelatinization enthalpy of the cross-linked starches decreased in proportion with irradiation dose. The melting temperatures of cross-linked starches gradually decreased and the temperature range for melting increased with an increase in irradiation dose.     

A mutation of the cellulose-synthase-like (CslF6) gene in barley (Hordeum vulgare L.) partially affects the β-glucan content in grains

The chemical induced barley mutant m351 was first selected for its low level of mixed-linkage (1–3,1–4) beta-D-glucan (MLG) in an experimental effort to search for barley lines with varied grain MLG contents. The MLG decrease in m351 was associated with increased levels of fructans and crude fiber, but maintained the same plant characteristics under field conditions. The mutation was mapped to a genetic locus flanked by two SSR markers, Bmag369 and Bmag564, on chromosome 7H. Molecular cloning of the CslF6 gene from the m351 line revealed the presence of a point mutation, causing a substitution of an alanine for threonine at position 849 in the amino acid sequence of the corresponding protein. The resultant protein retains some functionality and affects other components in the m351 grain. Those metabolic changes associated with MLG reduction in m351 is the first case reported of a partially functional CslF6 gene in cereal grains. The results contribute to better understanding of the functional effects of the CslF6 gene and the mutant has potential implications in grain end-use quality improvement.     

Studies on bioactivities of tea (Camellia sinensis L.) fruit peel extracts: Antioxidant activity and inhibitory potential against α-glucosidase and α-amylase in vitro

Tea fruit peel is the main byproduct during manufacture of tea seed oil. The great increase in tea seed oil production in recent years brings the challenge of finding application for tea fruit peel. The aims of this study were to obtain tea fruit peel extracts enriched with bioactive compounds by several solvent extraction methods and to evaluate their antioxidant activity and inhibitory potential against α-glucosidase and α-amylase in vitro. Flavonoids and phenolics were accumulated in ethyl acetate, butanol, and chloroform fractions, and these fractions possessed much better antioxidant activities, including scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) cation radicals (ABTS⁺), and reducing activity, compared with those of the 75% ethanol extract and water fraction. Moreover, the ethyl acetate, butanol, and chloroform fractions exhibited excellent inhibitory activity against α-glucosidase, much stronger than that of the positive control (acarbose). These fractions also showed mild inhibition on α-amylase activity.     

Are plant cell wall hydrolysing enzymes of saprobe fungi implicated in the biological control of the Verticillium dahliae pathogenesis?

We studied the possibility that the antagonistic saprobe fungi Coriolopsis rigida, Trametes versicolor, Fusarium lateritium, Penicillium chrysogenum and Verticillium dahliae-2379 may control V. dahliae disease through the inhibition of the activity of hydrolytic enzymes produced by the pathogen. These saprobe fungi were able to decrease the growth in vitro of V. dahliae. The exudates produced by these fungi seemed to be the main factor responsible for their antagonistic effect. The exudates of all the fungi tested had endopolymethylgalacturonase, endoglucanase and endoxyloglucanase activities. Exudates of P. chrysogenum and V. dahliae-2379 had higher hydrolytic activities than those of V. dahliae. The saprobe V. dahliae-2379 did not decrease the growth of the pathogen V. dahliae but decreased its harmful effect on the growth of tomato. However, no inhibition of the activities of the hydrolytic enzyme of the pathogenic V. dahliae by the no pathogenic V. dahliae-2379 or by the other saprobe fungi was observed. Nevertheless, the protection of tomato by the five antagonistic saprobe fungi against V. dahliae disease could be carried out by the induction of plant defense by the hydrolytic enzymes produced by these fungi.     

How many Orius laevigatus are needed for effective western flower thrips, Frankliniella occidentalis, management in sweet pepper?

Frankliniella occidentalis (Pergande) is a primary pest of greenhouse crops worldwide, in organic and integrated pest management control practices, Orius spp. are frequently released for thrips control. However, Orius spp. are relatively expensive to produce. More cost-efficient rearing systems and reduced release rate might reduce the expense. In these trials, we released Orius laevigatus (Fieber) at different rates with or without simultaneous release the predatory mite Amblyseius swirskii Athias-Henriot, another known thrips predator, which is less expensive to rear. There was no significant difference in the number of O. laevigatus recovered in which either 2 or 6 individuals were released per square meter, and there was no difference in thrips control among any of the release strategies using O. laevigatus, suggesting that a reduced release rate can maintain effective thrips control. There was no significant difference in the quality or quantity of the pepper yield between treatments in which either 2 or 6 Orius/m² or Orius plus A. swirskii were released.     

Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Towards an optimal process for gelatinisation and hydrolysis of highly concentrated starch–water mixtures with alpha-amylase from B. licheniformis

The enzymatic hydrolysis of starch is usually carried out with 30–35 w/w% starch in water. Higher substrate concentrations (50–70 w/w%) were reached by using a twin-screw extruder for gelatinisation and for mixing enzyme with gelatinised starch prior to enzymatic hydrolysis in a batch reactor. The aim of this study was to determine which parameters are important for gelatinisation of wheat starch and to investigate the effects of different extrusion conditions on the enzymatic hydrolysis. After extrusion, the degree of gelatinisation was measured. During hydrolysis, the carbohydrate composition, the dextrose equivalent (DE) and the alpha-amylase activity were measured. Gelatinisation measurements showed that mechanical forces lowered the temperature required for complete gelatinisation. During hydrolysis experiments, high DEs were observed even if starch was not completely gelatinised during extrusion. Due to high substrate concentrations, the residual alpha-amylase activity remained high throughout enzymatic hydrolysis, although high temperatures were used. Increased substrate concentrations did not affect the carbohydrate composition of the product. Furthermore, the time required for the batch hydrolysis step could be varied by choosing a different enzyme-to-substrate ratio. This article provides a basis for detailed optimisation of this process to develop an industrial-scale process at high substrate concentrations.     

Field assessment of coffee (Coffea arabica L.) cultivars in Meloidogyne exigua-infested or –free fields in Rio de Janeiro State,Brazil

A five-year field assessment of Meloidogyne exigua-susceptible and –resistant coffee cultivars was conducted in two fields, nematode-infested or –free, in Rio de Janeiro State, Brazil. The nematode-susceptible cultivars ‘IAC Catuaí Vermelho 144’, ‘Obatã’ and ‘Tupi’, own-rooted or grafted onto the nematode-resistant ‘IAC Apoatã 2258’, were compared in their vegetative development (plant height, number of plagiotropic branches and collar diameter) and productivity (during four harvests) with the nematode-resistant ‘Acauã’, ‘Catucaí 785/15’ and ‘Iapar 59’. In the infested field, the high nematode population drastically reduced the vegetative development and productivity of all cultivars, regardless of their nematode-resistance. This indicates that measures aimed to reduce the M. exigua population must be taken prior to planting any of the currently available resistant genotypes to avoid enormous yield losses. In the nematode-free field, the susceptible cultivars (own-rooted) achieved higher productivity, followed close by ‘Catucaí 785/15’ and ‘Iapar 59’. Therefore, these M. exigua-resistant cultivars should be recommended for nematode-free areas to prevent severe yield loss in the event of field infestation during the plantation's life span.     

Quantitative analyses of individual γ-Oryzanol (Steryl Ferulates) in conventional and organic brown rice (Oryza sativa L.)

γ-Oryzanol (steryl ferulates; SF) has been shown to be a major bioactive compound in rice. To determine the content of individual γ-oryzanols in brown rice by high performance liquid chromatography (HPLC), purification of individual SF for use as an external standard is required. Four main SF were isolated from a commercial γ-oryzanol mixture and identified as cycloartenyl ferulate (1), 24-methylenecycloartanyl ferulate (2), campesteryl ferulate (3), and sitosteryl ferulate (4) based on mass spectrometry and nuclear magnetic resonance spectroscopic data. The SF contents between conventional and organic brown rice were qualitatively determined by HPLC using SF isolated from a commercial γ-oryzanol mixture as the external standard. The total γ-oryzanol content (mg/100 g) in organic brown rice (65.6 ± 2.7) was slightly higher (P < 0.05) than that found in conventional brown rice (60.2 ± 1.8). The content (mg/100 g) of 1 (21.2 ± 0.9) and 4 (9.8 ± 0.4) in organic brown rice was higher (P < 0.05) than that observed in conventional brown rice (1, 18.2 ± 1.1; 4, 8.5 ± 0.3). However, the content of 2 and 3 in the conventional and organic brown rice samples did not differ significantly. These results indicate that the cultivation methods do significantly alter the γ-oryzanol content for conventional and organic brown rice.     

Effect of elevated carbon dioxide (CO2) on phenolic acids,flavonoids, tocopherols,tocotrienols, γ-oryzanol and antioxidant capacities of rice (Oryza sativa L.)

There have been no studies conducted with the objective of investigating the effect of elevated CO₂ concentrations ([CO₂]) on antioxidants in grains. Therefore, a two-year field experiment was conducted using open-top chambers with two levels of atmospheric CO₂ (375 and 550 μmol/mol) to evaluate their effects on rice grain antioxidants. Following exposure to high [CO₂], the total phenolic content of all rice milling fractions decreased (3%–18%), with the highest reduction in the brown rice for sinapic acid (167%), and in the white rice for p-hydroxybenzoic acid (100%). The total flavonoid content also decreased under elevated [CO₂] in all rice milling fractions (8%–14%), with apigenin (25%) being highly affected in the white rice, and tricin (12%) in the bran. The same trend was found for γ-oryzanol, with decreases of 35%, 32%, 25%, and 2% in the white rice, brown rice, husk, and bran, respectively. In the white and brown rices, tocopherols and tocotrienols were all lower under elevated [CO₂], with reductions larger for α-tocotrienol (69%), γ-tocotrienol (46%), and α-tocopherol (38%). Good correlations between antioxidant contents and DPPH radical scavenging capacities indicated that these decreases may be meaningful in the preventive ability of rice against free radical-mediated degenerative diseases.     

Does intercropping winter wheat (Triticum aestivum) with red fescue (Festuca rubra) as a cover crop improve agronomic and environmental performance? A modeling approach

The introduction of a living cover crop during a cash crop growth cycle (relay intercropping) and its maintenance after the cash crop harvest may help to preserve biodiversity, increase soil organic matter content and carbon sequestration and provide other ecosystem services, such as natural pest regulation or nutrient recycling, by increasing useful biotic interactions within the agroecosystem. We studied the impact of various approaches to manage a red fescue cover crop in a winter wheat crop in terms of light, water and nitrogen competition, using the STICS crop model adapted for intercropping. The STICS model for wheat/fescue intercropping was first evaluated on two years of experimental data obtained in the field. It gave satisfactory statistical results for the prediction of dry matter, leaf area index (LAI) and nitrogen accumulation in the two species, and for nitrogen and water dynamics in the soil. By simulating unmeasured variables, such as transpiration, the model improves our understanding of the performance of the intercrop in the field. For example, we showed that the intercropping system was more efficient that wheat grown as a sole crop, in terms of nitrogen accumulation and decreasing soil nitrogen levels before the leaching period. However, it also resulted in lower wheat yields. We then used the STICS model to compare four intercropping management scenarios differing in terms of the date of red fescue emergence, over 35 climatic years. We found that, in most climatic scenarios, the emergence of the fescue crop during the late tillering phase of the wheat crop gave the best compromise between wheat yield overall nitrogen accumulation and radiation interception.     

Modelling the β-amylase activity during red sorghum malting when Bacillus subtilis is used to control mould growth

Steeping in dilute alkaline (0.2% NaOH) followed by resteeping in biocontrol (starters of Bacillus subtilis S499) has been used during red sorghum malting. The effect of steeping and germination conditions has been described using 2 functions: a Weibull 4-parameter model combined with a General Linear Model with Logarithm Link with significant goodness. Steeping conditions (combined use of NaOH and B. subtilis S499) affects the synthesis capacity of grain: when B. subtilis culture used in the steeping step is diluted, ln α increases, suggesting a loss of treatment efficacy. The germination temperature affects the β-amylase synthesis rate during the induction phase: the germination temperature increase is accompanied by a decrease of the β-amylase synthesis rate. During the repression phase of β-amylase synthesis, the effect of malting conditions was found to taper.     

Indirect changes associated with a selection program for increased seed-yield in wild species of Lesquerella (Brassicaceae): Are we developing a phenotype opposite to the expected ideotype?

Seed-yield stability, frequently associated with drought-tolerance strategies, is one of the main breeding objectives for the development of crops for semi-arid mediterranean-type environments. Since breeding of new industrial crops targeted for arid lands is not appreciably different from that of traditional crops, higher yield is achieved by increased harvest-index, at the cost of losing traits associated with drought-tolerance and reduced seed-yield stability. Using Lesquerella as a model we compared selected and unselected accessions of annual (L. gracilis and L. angustifolia) and perennial (L. pinetorum and L. mendocina) genotypes grown in field experiments in Patagonia, Argentina. Our objective was to assess the effects of breeding for increased seed-yield on traits related to the main characteristics that define the most common ideotype for mediterranean-type environments: early vigor, conservative growth strategy post-anthesis and reserves storage. Our specific question was: Have any of the attributes associated with seed-yield stability been indirectly selected during the domestication process? Our results show that these characteristics were reduced or lost, in selected lines compared to their wild relatives. Early vigor was lower in selected accessions and was associated mainly with reduced relative growth rate and CO₂ assimilation. During the reproductive period the growth strategy was changed by selection towards a non-conservative and more acquisitive resource use strategy. Traits associated with this strategy were linked to higher water use efficiency and growth capacity (higher CO₂ assimilation rate, specific leaf area, and leaf allocation), but also with loss of structural adaptation to low resource environments (i.e. low specific leaf area), an increase in nutrient and water demands, and reduced nutrient use efficiency. Carbohydrates accumulation pre-anthesis was lower in selected accessions of all four species, and also, just in perennials we found lower reserves storage post-anthesis. These changes in the pattern of carbohydrates accumulation could be associated to lower seed-yield stability due to the loss of buffer capacity linked with the use of pre-anthesis reserves for seed filling. On the other hand, in perennial species lower reserves storage after seed harvest could reduce plant longevity and survival. We conclude that indirect changes occurred during the preliminary domestication of both annual and perennial species of Lesquerella used in our experiment. These changes were against those required if these species were to be developed as crops for semi-arid, mediterranean environments and should result in low seed-yield stability.