Scielo RSS <![CDATA[Electronic Journal of Biotechnology]]> vol. 17 num. 5 lang. pt <![CDATA[SciELO Logo]]> <![CDATA[<strong>Physicochemical and nutritional alterations induced by two-spotted spider mite infestation on strawberry plants</strong>]]> Background Strawberry is a pseudofruit mainly cultivated in temperate climate regions. Considering its high levels of ascorbic acid and phenolic compounds, the consumption of strawberry fruit can be beneficial to health. The Brazilian strawberry production revolves around 3000 tons per year, significantly influencing the food market and generating income to farmers. However, this production can be partially impaired by two-spotted spider mite (TSSM) Tetranychus urticae Koch infestations, due to decreases in the quality and quantity of fruit. Since there are no data in the literature about alterations caused by TSSM infestation in strawberry plants, our work aimed towards evaluating nutritional and physicochemical parameters of TSSM-infested strawberry plants, along with the related chemical treatment (CT) (acaricide) or biological treatment (predatory mite Phytoseiulus macropilis Banks). Results Strawberry fruit from TSSM-infested plants present the highest levels of acidity and exhibit low levels of anthocyanin and phenolic compounds, while fruit from TSSM-infested plants + biological control using predatory mite shows high levels of soluble solids, phenolic compounds and ascorbic acid, along with a high soluble solid content/titratable (SSC/TA) acidity ratio, which indicates high quality fruit. Conclusions Our results suggest that TSSM infestation decreases fruit quality and that the biological control of TSSM using a predatory mite is a suitable alternative to organic production, since the presence of predatory mite does not affect fruit quality and development. <![CDATA[<strong>Impact of simulated acid rain on soil microbial community function in Masson pine seedlings</strong>]]> Background Accompanying its rapid economic development and population growth, China is the world's third largest acid rain region, following Europe and North America. The effects of acid rain on forest ecosystem were widely researched, including the growth, the nutrient of the leaf and soil, and so on. However, there are few reports about the effects of acid rain on the soil microbial diversity. This study investigated the effects of acid rain on soil microbial community function under potted Masson pine seedlings (Pinus massoniana Lamb). Results After 7 months of treatment with simulated acid rain, the low acid load treatment (pH 5.5) stimulated soil microbial activity, and increased soil microbial diversity and richness, while the higher levels of acid application (pH 4.5, pH 3.5) resulted in lower soil microbial activity and had no significant effects on soil microbial diversity and richness. Principal component analysis showed that there was clear discrimination in the metabolic capability of the soil microbial community among the simulated acid rain and control treatments. Conclusion The results obtained indicated that the higher acid load decreased the soil microbial activity and no effects on soil microbial diversity assessed by Biolog of potted Masson pine seedlings. Simulated acid rain also changed the metabolic capability of the soil microbial community. <![CDATA[<strong>Prolyl endopeptidase - Optimization of medium and culture conditions for enhanced production by </strong><em><b>Lactobacillus acidophilus</b></em>]]> Background Lactic acid bacteria are able to reduce the immunoreactivity of proteins of cereal grains during wheat dough fermentation or may be a source of proteolytic preparations added during bread making. The key enzyme in prolamin degradation is prolyl endopeptidase. This study was aimed at optimizing the composition of a culture medium and culture conditions that would enhance the synthesis of intracellular prolyl endopeptidase (PEP) by Lactobacillus acidophilus 5e2. Results The application of Plackett-Burman screening plans enabled demonstrating that the concentration of a nitrogen source in the culture and the initial pH value of the culture medium were significant for PEP synthesis. Further optimization conducted with the method of central composite designs (CCD) confirmed both the linear and square impact of nitrogen concentration and initial pH value of the culture medium on PEP production. In turn, the response surface method (RSM) allowed determining the optimal nitrogen concentration and pH value at 26.88 g/l and pH 4.85, respectively. Conclusions Validation of the resultant model enabled over 3-fold increase in the quantity of the synthesized enzyme. <![CDATA[<strong>Establishment of a pheasant (</strong><em><b>Phasianus colchicus</b></em><strong>) spermatogonial stem cell line for the production of interspecies germ line chimeras</strong>]]> Background Spermatogonial stem cells (SSCs) are important for the production of interspecies germ line chimeras. The interspecies germ cell transfer technique has been suggested as a way to conserve endangered birds. Our objective was to develop a technique for restoring endangered birds by developing interspecies germ line chimeras between pheasant (Phasianus colchicus) and chicken (Gallus gallus) with SSCs. Results SSCs were isolated from the surgically removed testis of a pheasant. Growth conditions for pheasant SSCs were established by co-culturing STO (SIM mouse embryo-derived thioguanine and ouabain resistant) cells and pheasant SSCs. The colony-forming cells divided and proliferated stably to yield an established SSC line. Pheasant SSCs showed strong reactivity for GDNF family receptor alpha1 (GFRa1) marker. Finally, production of germ line chimeras was attempted by transferring pheasant SSCs into recipient embryos. Although final embryo survival was 5.6% (20/354), the initial survival rate was 88% (312/354). To measure the percent transfer of donor SSC to gonads, the pheasant SSCs were labeled with PKH 26 fluorescent dye. We observed 30% donor cells and 9.48% c-kit/CD117-positive cells in the gonads of recipient chickens. Donor SSCs were thus stably engrafted in the recipient gonads. Conclusions This study showed that SSCs can be used as a tool for the conservation of endangered birds and the production of germ line chimeras. Our findings yield insights into how we may use the pheasant spermatogonial stem cell line for efficient production of interspecies germ line chimeras and ultimately, to the restoration of endangered birds. <![CDATA[<strong>Pleiotropic effects of the </strong><em><b>sdw1</b></em><strong> locus in barley populations representing different rounds of recombination</strong>]]> Background In the present study populations, representing different rounds of recombination were used for the analysis of phenotypic effects associated with the sdw1/denso locus. Other studies have mostly focused only on one type of population. Many different QTLs mapped at the same position as the sdw1/denso locus may indicate a pleiotropy of this gene or a tight linkage between genes conditioning quantitative traits. To date, results of studies have not unequivocally proven either of these two phenomena. Results Both breeding and molecular mapping experiments were undertaken to examine 200 single seed descent (SSD) and 60 doubled haploid (DH) lines obtained from the Maresi (with a semi-dwarfing gene) and Pomo cross combination. They were evaluated for the type of juvenile growth habit and certain agronomic traits were measured after harvesting. The estimates of mean values, standard errors and significance of effects were analyzed. In terms of the analyzed characteristics, the greatest variability was obtained for genotypes with the prostrate growth habit. Microsatellite markers (SSR) were also used to identify co-segregation with the sdw1/denso locus and Bmag0013, Bmag0877, Bmag0306b markers were linked the closest. A partial linkage map of chromosome 3H with the sdw1/denso semi-dwarfing gene was constructed and QTLs were identified. Conclusions Our experiments confirmed the impact of the semi-dwarfing gene on plant height, heading and flowering date both in SSD and DH populations, which may indicate pleiotropy. Moreover, a partial linkage between sdw1/denso locus and grain weight per spike and 1000-grain weight was found in the SSD population. <![CDATA[<strong>Construction of a eukaryotic expression vector for pEGFP-FST and its biological activity in duck myoblasts</strong>]]> Background Follistatin (FST), a secreted glycoprotein, is intrinsically linked to muscle hypertrophy. To explore the function of duck FST in myoblast proliferation and differentiation, the pEGFP-FST eukaryotic expression vector was constructed and identified. The biological activities of this vector were analyzed by transfecting pEGFP-FST into cultured duck myoblasts using Lipofectamine™ 2000 and subsequently determining the mRNA expression profiles of FST and myostatin (MSTN). Results The duck pEGFP-FST vector was successfully constructed and was confirmed to have high liposome-mediated transfection efficiency in duck myoblasts. Additionally, myoblasts transfected with pEGFP-FST had a higher biological activity. Significantly, the overexpression of FST in these cells significantly inhibited the mRNA expression of MSTN (a target gene that is negatively regulated by FST). Conclusions The duck pEGFP-FST vector has been constructed successfully and exhibits biological activity by promoting myoblast proliferation and differentiation in vitro. <![CDATA[Analysis of gene expression profiles in response to <em><b>Sporisorium reilianum</b></em><strong> f. sp. zeae in maize (</strong><em><b>Zea may</b></em><strong>s L.)</strong>]]> Background Head smut of maize, which is caused by Sporisorium reilianum f. sp. zeae (Kühn), is a serious disease in maize. In order to reveal the molecular mechanism of the resistance to head smut in maize, a microarray containing ~ 14,850 probes was used to monitor the gene expression profiles between a disease resistant near isogenic line (NIL) and a highly susceptible inbred line after S. reilianum was injected with an artificial inoculation method. Results Levels of expression for 3,532 genes accounting for 23.8% of the total probes changed after inoculation. Gene Ontology analysis revealed that the differentially expressed genes participated in physiological and biochemical pathways. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that plant-pathogen interaction, natural killer cell mediated cytotoxicity and benzoxazinoid biosynthesis pathways play important roles in resistance to head smut. Three head smut resistance-related candidate genes, CLAVATA1, bassinosteroid insensitive 1-associated receptor kinase 1 and LOC100217307 with leucine-rich repeat (LRR) conserved domains were identified, each of which is in maize mapping bin 2.09, a region previously shown to include a major QTL for head smut resistance. Furthermore, LOC100217307 was validated by quantitative real-time (qRT)-PCR inferring that this gene may be involved in the resistance to head smut of maize. Conclusions This study provided valuable information for cloning, functional analysis and marker assisted breeding of head smut resistance genes. <![CDATA[<strong>Plant prebiotics and human health</strong>: <strong>Biotechnology to breed prebiotic-rich nutritious food crops</strong>]]> Microbiota in the gut play essential roles in human health. Prebiotics are non-digestible complex carbohydrates that are fermented in the colon, yielding energy and short chain fatty acids, and selectively promote the growth of Bifidobacteria and Lactobacillae in the gastro-intestinal tract. Fructans and inulin are the best-characterized plant prebiotics. Many vegetable, root and tuber crops as well as some fruit crops are the best-known sources of prebiotic carbohydrates, while the prebiotic-rich grain crops include barley, chickpea, lentil, lupin, and wheat. Some prebiotic-rich crop germplasm have been reported in barley, chickpea, lentil, wheat, yacon, and Jerusalem artichoke. A few major quantitative trait loci and gene-based markers associated with high fructan are known in wheat. More targeted search in genebanks using reduced subsets (representing diversity in germplasm) is needed to identify accessions with prebiotic carbohydrates. Transgenic maize, potato and sugarcane with high fructan, with no adverse effects on plant development, have been bred, which suggests that it is feasible to introduce fructan biosynthesis pathways in crops to produce health-imparting prebiotics. Developing prebiotic-rich and super nutritious crops will alleviate the widespread malnutrition and promote human health. A paradigm shift in breeding program is needed to achieve this goal and to ensure that newly-bred crop cultivars are nutritious, safe and health promoting. <![CDATA[<strong>On the fermentative behavior of auxotrophic strains of </strong><em><b>Saccharomyces cerevisiae</b></em>]]> Background The selection of new yeast strains could lead to improvements in bioethanol production. Here, we have studied the fermentative capacity of different auxotrophic mutants of Saccharomyces cerevisiae, which are routinely used as hosts for the production of heterologous proteins. It has recently been found that these strains exhibit physiological alterations and peculiar sensitivities with respect to the parental prototrophic strains from which they derive. In this work the performance of auxotrophic S. cerevisiae CEN.PK strains was compared to the corresponding prototrophic strain, to S. cerevisiae T5bV, a strain isolated from grape must and to another auxotrophic strain, S. cerevisiae BY4741. Results The results indicate that the fermentative capacity of strains grown in 2% glucose was similar in all the strains tested. However, in 15% initial glucose, the auxotrophic strains exhibited a more than doubled ethanol yield on biomass (10 g g- 1dw) compared to the prototrophic strains (less than 5 g g- 1dw). Other tests have also evidenced that in medium depletion conditions, ethanol production continues after growth arrest. Conclusions The results highlight the capacity of auxotrophic yeast strains to produce ethanol per mass unit, in a higher amount with respect to the prototrophic ones. This leads to potential applications for auxotrophic strains of S. cerevisiae in the production of ethanol in both homogeneous and heterogeneous phases (immobilized systems). The higher ethanol yield on biomass would be advantageous in immobilized cell systems, as a reduced yeast biomass could greatly reduce the mass transfer limitations through the immobilization matrix.