EpiHRMAssay, in tube and in silico combined approach for the scanning and epityping of heterogeneous DNA methylation

Reliable and cost-effective assays with adequate sensitivity are required to detect the DNA methylation profile in plants for scientific and industrial purposes. The proposed novel assay, named EpiHRMAssay, allows to quantify the overall methylation status at target loci and to enable high-throughput analyses. It combines in tube High Resolution Melting Analysis on bisulphite-treated templates with the in silico prediction of the melting profile of virtual epialleles using uMELTSM software. The predicted melting temperatures (Tm-s) of a set of epialleles characterized by different numbers of methylated cytosines (#mC) or different mC configurations were obtained and used to build calibration models, enabling the quantification of methylation in unknown samples using only the in tube observed melting temperature (Tm-o). EpiHRMAssay was validated by analysing the promoter region of CMT3, DDM1, and ROS1 genes involved in the regulation of methylation/demethylation processes and chromatin remodelling within a population of peach plants. Results demonstrate that EpiHRMAssay is a sensitive and reliable tool for locus-specific large-scale research and diagnostic contexts of the regulative regions of genes, in a broad range of organisms, including mammals. EpiHRMAssay also provides complementary information for the assessment of heterogeneous methylation and can address an array of biological questions on epigenetic regulation for diversity studies and for large-scale functional genomics

Vertical Greenery as Natural Tool for Improving Energy Efficiency of Buildings

The European Construction Sector Observatory outlined that green building envelopes as green roofs and walls contribute to the reduction of energy demand and CO2 emissions due to the air conditioning in summer periods, and the mitigation of heat islands in urban areas. For this reason, the understanding about the contribution of urban greening infrastructures on buildings to sustainable energy use for air conditioning is urgent. This paper focuses on the analysis of a vertical surface provided with a Parthenocissus quinquefolia (L.) Planch., a winter deciduous species, as green cover of a building, assessing the reduction of the solar radiation energy absorbed by the facade and, consequently, the heat flux (HF) transmitted into the internal ambient. This research shows that, in July, surface temperatures (STs) on the vegetated facade were up to 13 degrees C lower than on the unvegetated (bare) facade. Under the climate and environmental conditions of the green wall located at ENEA Casaccia Research Center, a saving of 2.22 and 1.94 kWh(e)/m(2) , respectively in 2019 and 2020, for the summer cooling electricity load, was achieved. These energy reductions also allowed the saving of 985 and 862 g CO2/m(2) emissions, respectively, in 2019 and 2020. Ultimately, a green factor named K-v* was also elaborated to evaluate the influence of vegetation on the STs as well as on HFs transmitted into the indoor ambient and adapted to the case of a detached vertical green cover. Measurements of K-v* factor lasting three years showed the suitability of this index for defining the shading capacity of the vegetation on the building facade surfaces, which can be used to predict thermal gains and effects in a building endowed of a vertical green system

Exogenous miRNAs from Moringa oleifera Lam. recover a dysregulated lipid metabolism

A balanced diet is critical for human health, and edible plants play an important role in providing essential micronutrients as well as specific microRNAs (miRNAs) that can regulate human gene expression. Here we present the effects of Moringa oleifera (MO) miRNAs (mol-miRs) on lipid metabolism. Through in silico studies we identified the potential genes involved in lipid metabolism targeted by mol-miRs. To this end, we tested the efficacy of an aqueous extract of MO seeds (MOES), as suggested in traditional African ethnomedicine, or its purified miRNAs. The biological properties of MO preparations were investigated using a human derived hepatoma cell line (HepG2) as a model. MOES treatment decreased intracellular lipid accumulation and induced apoptosis in HepG2. In the same cell line, transfection with mol-miRs showed similar effects to MOES. Moreover, the effect of the mol-miR pool was investigated in a pre-obese mouse model, in which treatment with mol-miRs was able to prevent dysregulation of lipid metabolism

MicroRNA from moringa oleifera : identification by high throughput sequencing and their potential contribution to plant medicinal value

Moringa oleifera is a widespread plant with substantial nutritional and medicinal value. We postulated that microRNAs (miRNAs), which are endogenous, noncoding small RNAs regulating gene expression at the post-Transcriptional level, might contribute to the medicinal properties of plants of this species after ingestion into human body, regulating human gene expression. However, the knowledge is scarce about miRNA in Moringa. Furthermore, in order to test the hypothesis on the pharmacological potential properties of miRNA, we conducted a high-Throughput sequencing analysis using the Illumina platform. A total of 31,290,964 raw reads were produced from a library of small RNA isolated from M. oleifera seeds. We identified 94 conserved and two novel miRNAs that were validated by qRT-PCR assays. Results from qRT-PCR trials conducted on the expression of 20 Moringa miRNA showed that are conserved across multiple plant species as determined by their detection in tissue of other common crop plants. In silico analyses predicted target genes for the conserved miRNA that in turn allowed to relate the miRNAs to the regulation of physiological processes. Some of the predicted plant miRNAs have functional homology to their mammalian counterparts and regulated human genes when they were transfected into cell lines. To our knowledge, this is the first report of discovering M. oleifera miRNAs based on highthroughput sequencing and bioinformatics analysis and we provided new insight into a potential cross-species control of human gene expression. The widespread cultivation and consumption of M. oleifera, for nutritional and medicinal purposes, brings humans into close contact with products and extracts of this plant species. The potential for miRNA transfer should be evaluated as one possible mechanism of action to account for beneficial properties of this valuable species

How soil ecological intensification by means of cover crops affects nitrogen use efficiency in pepper cultivation

Ecological intensification, based on agricultural practices that promote ecosystem services, has been recently proposed to match crop yield and environmental concerns. Two-year experiments were conducted in a Mediterranean environment. The treatments were: (i) four intensification levels (common vetch (CV), ryegrass (RG), bare soil without Nitrogen (N) fertilization (Control-N0) and with 100 kg ha-1 ofNfertilization (Control-N100) applied during pepper cultivation), and(ii) two soil tillage [soil tillage at 15 cm and 30 cm of soil depth (ST-15 and ST-30, respectively)]. The field experiment was disposed in a randomized block design with three replications. Cover crop, soil samples, and pepper samples were collected for analysis. Soil available nitrogen increased after soil tillage, especially in CV, which showed the highest fruit yield. The reduced soil N availability in RG decreased fruit yield and N uptake. The agro-physiological efficiency of pepper was similar in common vetch and Control-N100, while it was low in ryegrass. However, the adoption of RG increased the soil organic matter more than both control treatments, which, in turn, caused a depletion of soil organic matter. Moreover, reduced tillage practices for green manuring that both cover crops arepreferable to reduce external inputs in terms of fuel saving and farming operations

Physiological dissection of blue and red light regulation of apical dominance and branching in M9 apple rootstock growing in vitro

This paper presents the results of the interaction of red light (R) with blue light (B), applied to shoots of M9 apple (Malus pumila paradisiaca Schmid) rootstock, on the regulation of stem growth, apical dominance and branching. These results are compared with the active form of phytochromes (PHYs) under monochromatic and dichromatic light treatments. The inhibition of internode elongation was determined by PHY photoequilibrium, with the additional effect of B, by means of a separate photoreceptor. The development of phytomers appeared to be primarily due to the active form of PHY, with a marginal effect from B. Shoot growth, which combines internode elongation and development of the phytomer, was highest under R and lowest under B and far red light (FR), showing the largely positive role of PHY photoequilibrium. Under FR, reduced stem elongation was due to the very small number of phytomers formed. Apical dominance was inhibited, while branching was increased under R, corresponding to the highest values of PHY photoequilibrium determined. Apical dominance was increased and branching was reduced by B, indicating a complex interaction between PHY and B receptor. In the shoot cluster system, photomorphogenic behavior was dependent on the time of exposure to the different light qualities. The information gained from the study will be helpful in improving the set up of in vitro growth light conditions, and in providing useful insights into research of the development of the woody plant canopy, an important factor in ecological plant communities. © 2008 Elsevier GmbH. All rights reserve