In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus–oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/μl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/μl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/μl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.     

Near-Infrared Spectroscopy Coupled with Chemometrics Tools: A Rapid and Non-Destructive Alternative on Soil Evaluation

Near-infrared (NIR) spectroscopy is a rapid, non-destructive and accurate technique for analyzing a wide variety of samples, thus, the growing interest of using this technique in soil science. The objective of this study was to evaluate the potential of NIR spectroscopy to predict organic carbon (OC), total nitrogen (TN), available phosphorus (P) and available potassium (K) in the soil. NIR spectra from 20 cm³ of soil samples were acquired on the range of 750 to 2500 nm in diffuse reflectance mode, resolution of 16 cm^?1 and 64 scans. Eight models of calibration/validation were constructed. Calibration and validation models showed that the predictive potential of NIR varied with the specific soil property (OC, TN, P and K) under evaluation and according to the methodology employed in the model construction (cross-validation or test set). Good prediction models were obtained for OC and TN content based on the statistical parameters. Test set methodology was able to predict soil OC, TN, P, and K better than cross-validation methodology.     

Biofortification of soybean grains with foliar application of Li sources

Abstract

Biofortification of soybean grains with lithium (Li) is a strategy to improve a food with high social acceptance, in order to promote health benefits. The aim of this work was to evaluate the development of plants and the production of biofortified soybeans with two Li sources. The experiment was conducted in a completely randomized design with five replicates. Ten treatments were obtained in a 2x5 factorial scheme, comprising two sources of Li (LiOH - Li hydroxide and Li₂SO₄ - Li sulfate) and five doses (0, 30, 60, 90 and 120?mg kg^?1) in stages V4 and R1. The supply of Li promoted significant effects (p?≤?0.05) on the morphology, yield components and nutritional status of soybean plants. The highest grain yield was obtained with the use of Li₂SO₄ at the estimated dose of 45.7?mg kg^?1. The plants accumulated Li between 8.00 and 11.20?mg kg ^?1, respectively, with the use of Li₂SO₄ and LiOH. In the grains, the highest concentrations of Li were obtained with the application of 120?mg kg^?1. Li₂SO₄ and LiOH are good sources for biofortification of soybean grains.     

Optimization of conditions for anthocyanin hydrolysis from red wine using response surface methodology (RSM)

Optimization of conditions for anthocyanin hydrolysis from red wine was investigated using response surface methodology. The aglycon forms of the anthocyanins were quantified by high-performance liquid chromatography with diode array detection. The combined effects of three independent variables, HCl amount, heating temperature, and hydrolysis time, were studied using a 2(3) full-factorial central composite design. Anthocyanin hydrolysis yield depended mainly on the heating temperature and time of hydrolysis. HCl amount was the factor that least influenced the hydrolysis of anthocyanins. From experimental results, the maximum yield of anthocyanidins was reached with 9.8 mL of HCl (32% v/v), a heating temperature of 166.2 °C, and a hydrolysis time of 46.6 min. Five anthocyanidins, namely, delphinidin, cyanidin, petunidin, peonidin, and malvidin, were quantified in red wine. The reliability of the method was confirmed by recovery experiments, performed under optimal conditions. Recoveries indicated that anthocyanidins resisted the hydrolysis conditions.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Response of stevia to nitrogen fertilization and harvesting regime in northeastern Portugal

The adaptation of stevia to the growing conditions of NE Portugal is assessed, including the tolerance of this species to cold temperatures, and the potential to produce biomass when grown as an annual crop and when subjected to various nitrogen (N) rates and two harvesting regimes. Almost all the plants died during the winter of 2014 (minimum temperatures peaked at ?8.0°C), making it necessary to replant the crop the following spring. With the best cutting regime (double cut) and N rate (150 kg N ha^?1), 1514.4 and 2390.0 kg ha^?1 of dry leaves were produced, respectively, in 2014 and 2015. Leaf chlorophyll concentrations estimated by the SPAD (Soil and Plant Analysis Development)-502 chlorophyll meter and a NDVI (Normalized Difference Vegetation Index) carried out by the Field Scout CM 1000 spectroradiometer showed significant differences among N rates, proving to be good indicators of plant N nutritional status. Based on the leaf analysis, provisional sufficiency ranges for N are proposed, namely 25–35 g kg^?1 for mid-summer and 15–25 g kg^?1 for early autumn. The fluorescence of chlorophyll a and the transient fluorescence intensity performed by the OS-30p+ fluorometer failed to show any stress induced by no-N control treatments in comparison to N-treated plants.     

Bulk density by Proctor test as a function of texture for agricultural soils in Maputo province of Mozambique

Soil degradation processes may be of various kinds, including soil compaction. The present study was carried out with the objective of assessing the sensitivity of agricultural or recently abandoned soils in Maputo province of Mozambique to compaction. The assessment is based on the maximum of bulk density attained using the Proctor test (MBD).

In this study the soil texture is expressed by silt plus clay (S + C) or clay (C). The relations between the soil texture and MBD, and between soil texture and critical water content (CWC—soil water at which MBD is attained) were determined. Selected soils range from 10 to 74% of S + C and 9 to 60% of C.

The results suggest there is a relationship between the considered parameters, being that between S + C and MBD or CWC, the best. For MBD the relationship is represented by two quadratic equations with the boundary in between these being a S + C value of 25% and C value of 20%.

Based on the obtained results, one can conclude that the selected parameters may be a useful basis for estimation of the sensitivity to compaction of the Maputo province's soils. It is recommended that similar studies be carried out for soils under forest land and for soil of other provinces to establish the national physical degradation hazard as a function of soil parameters determined routinely and at low cost. The suggested parameters are texture and soil organic matter (SOM).