Vitamin A deficiency is a public health problem in a large number of countries. Biofortification of major staple crops (wheat [Triticum aestivum], rice [Oryza sativa], maize [Zea mays], and potato [Solanum tuberosum]) with b-carotene has the potential to alleviate this nutritional problem. Previously, we engineered transgenic "Golden" potato tubers overexpressing three bacterial genes for b-carotene synthesis (CrtB, CrtI, and CrtY, encoding phytoene synthase, phytoene desaturase, and lycopene b-cyclase, respectively) and accumulating the highest amount of b-carotene in the four aforementioned crops. Here, we report the systematic quantitation of carotenoid metabolites and transcripts in 24 lines carrying six different transgene combinations under the control of the 35S and Patatin (Pat) promoters. Low levels of B-I expression are sufficient for interfering with leaf carotenogenesis, but not for b-carotene accumulation in tubers and calli, which requires high expression levels of all three genes under the control of the Pat promoter. Tubers expressing the B-I transgenes show large perturbations in the transcription of endogenous carotenoid genes, with only minor changes in carotenoid content, while the opposite phenotype (low levels of transcriptional perturbation and high carotenoid levels) is observed in Golden (Y-B-I) tubers. We used hierarchical clustering and pairwise correlation analysis, together with a new method for network correlation analysis, developed for this purpose, to assess the perturbations in transcript and metabolite levels in transgenic leaves and tubers. Through a "guilt-by-profiling" approach, we identified several endogenous genes for carotenoid biosynthesis likely to play a key regulatory role in Golden tubers, which are candidates for manipulations aimed at the further optimization of tuber carotenoid content.
The absence of toxic and genotoxic effects of the hydrolysates prepared by the three hydrolytic processes suggests that they do not negatively affect eukaryotic cells and soil ecosystems and that they can be used in conventional and organic farming as an important nitrogen source derived from otherwise highly polluting by-products.
Massive sequencing of fungal communities showed that climatic factors, followed by edaphic and spatial variables, are feasible predictors of fungal richness and community composition. This study, based on a long-term field experiment with tillage and no-tillage management since 1995 and with a crop rotation introduced in 2009, confirmed that tillage practices shape soil properties and impact soil fungal communities. Results highlighted higher biodiversity of saprotrophic fungi in soil sites with low disturbance and an inverse correlation between the biodiversity of ectomycorrhizal and saprotrophic fungi. We speculated how their mutual exclusion could be due to a substrate-mediated niche partitioning or by space segregation. Moreover, where the soil was ploughed, the species were evenly distributed. There was higher spatial variability in the absence of ploughing, with fungal taxa distributed according to a small-scale pattern, corresponding to micro-niches that probably remained undisturbed and heterogeneously distributed. Many differentially represented OTUs in all the conditions investigated were unidentified species or OTUs matching at high taxa level (i.e., phylum, class, order). Among the fungi with key roles in all the investigated conditions, there were several yeast species known to have pronounced endemism in soil and are also largely unidentified. In addition to yeasts, other fungal species emerged as either indicator of a kind of management or as strongly associated with a specific condition. Plant residues played a substantial role in defining the assortment of species.
<p>This paper shows the results of the monitoring carried out in three hilly farms of the MONACO project in order to verify the effectiveness of the Standard 1.1 <sub>(commitment a)</sub> (temporary ditches) and Standard 1.2 <sub>(commitment g)</sub> (Vegetation cover throughout the year in set-aside land) in the reduction in soil erosion, contained in Rule 1: ‘minimum land management that meets specific conditions’ of the decree Mipaaf 2009 and following modifications, until the recent decree No. 180 of January 23, 2015. In addition, the assessment of the competitiveness gap was done. That is the evaluation of the additional costs borne by the beneficiary of the single payment determined from agronomic commitments. Monitoring has also compared the erosion actually observed in the field with that predicted by RUSLE model (Revised Universal Soil Loss Equation) (Renard et al., 1997) in the two situations: with and without the presence of temporary ditches, i.e. assuming Factual (compliance rules) and in that Counterfactual (infringement). This comparison was made in view of the fact that the RUSLE model was chosen by the 'European Evaluation Network for Rural Development (EEN, 2013) as a forecasting tool for the quantification of' Common Indicator ‘soil erosion by water’. The results of soil erosion survey carried out by using a new UAV-GIS methodology on two monitoring farms in two years of observations have shown that temporary ditches were effective in decreasing erosion, on average, by 42.5%, from 36. 59 t ha<sup>-1</sup> to 21.05 t ha<sup>-1</sup> during the monitoring period. It was also evaluated the effectiveness of grass strips (at variance with the commitment of temporary ditches). The results showed a strong, highly significant, reduction in erosion by about 35% times respect soil erosion observed in bare soil and also a significant reduction in the volume of runoff water. With regard to Standard 1.2 <sub>(commitment g)</sub> the statistical analysis shows a strong and highly significant decrease in the erosion due to the vegetation cover of the soil compared to bare soil. The economic competitiveness gap of Standard 1.1<sub>(commitment a)</sub> stood at € 4.07±1.42 € ha<sup>-1</sup> year<sup>-1</sup>, while CO<sub>2</sub> emissions due to execution of temporary ditches was 2.58 kg ha<sup>-1</sup>year<sup>-1</sup>. As for the Standard 1.2 <sub>(commitment g) </sub>the average differential competitiveness gap amounted to 50.22±13.7 € ha<sup>-1</sup> year<sup>-1</sup> and an output of CO<sub>2</sub> equal to 31.52 kg ha<sup>-1</sup> year.</p>
The alteration of the organic matter (OM) and the composition of bacterial community in microbial fuel cells (MFCs) supplied with soil (S) and a composted organic fertilizer (A) was examined at the beginning and at the end of 3 weeks of incubation under current-producing as well as no-current-producing conditions. Denaturing gradient gel electrophoresis revealed a significant alteration of the microbial community structure in MFCs generating electricity as compared with no-current-producing MFCs. The genetic diversity of cultivable bacterial communities was assessed by random amplified polymorphic DNA (RAPD) analysis of 106 bacterial isolates obtained by using both generic and elective media. Sequencing of the 16S rRNA genes of the more representative RAPD groups indicated that over 50.4% of the isolates from MFCs fed with S were Proteobacteria, 25.1% Firmicutes, and 24.5% Actinobacteria, whereas in MFCs supplied with A 100% of the dominant species belonged to γ-Proteobacteria. The chemical analysis performed by fractioning the OM and using thermal analysis showed that the amount of total organic carbon contained in the soluble phase of the electrochemically active chambers significantly decreased as compared to the no-current-producing systems, whereas the OM of the solid phase became more humified and aromatic along with electricity generation, suggesting a significant stimulation of a humification process of the OM. These findings demonstrated that electroactive bacteria are commonly present in aerobic organic substrates such as soil or a fertilizer and that MFCs could represent a powerful tool for exploring the mineralization and humification processes of the soil OM.
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