Deleterious Effects of Progestagen Treatment in VEGF Expression in Corpora Lutea of Pregnant Ewes

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri‐implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen‐treated group showed smaller corpora lutea than cloprostenol‐treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen‐treated group, which even showed a decrease between days 11 and 13. In progestagen‐treated sheep, VEGF expression in granulosa‐derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.     

Simulating in situ ammonia volatilization losses in the North China Plain using a dynamic soil‐crop model

Ammonia (NH₃) volatilization is an important N loss pathway in intensive agriculture of the North China Plain (NCP). Simulation models can help to assess complex N and water processes of agricultural soil–crop systems. Four variations (Var) of a sub‐module for the deterministic, process‐based HERMES model were implemented ranging from simple empirical functions (Var 3 and 4) to process‐oriented approaches (Var 1 and 2) including the main processes of NH₃ volatilization, urea hydrolysis, nitrification from ammonium‐based N fertilizer, and changes in soil solution pH. Ammonia volatilization, plant growth, and changes in ammonium and nitrate pools in the soil over several winter wheat–summer maize double‐crop rotations at three locations in the NCP were simulated. Results were calibrated with two data sets (Dongbeiwang 1, Shunyi) and validated using two data sets (Dongbeiwang 2, Quzhou). They showed that the ammonia volatilization sub‐module of the HERMES model worked well under the climatic and soil conditions of N China. Although the simpler equations, Var 3 and 4, showed lower deviations to observed volatilization across all sites and treatments with a mean absolute error (MAE) of 1.8 and 1.4 in % of applied N, respectively, compared to process‐oriented approaches, Var 1 and 2, with a MAE of 2.2 and 1.9 in % of applied N, respectively. Environmental conditions were reflected better by the process‐oriented approaches. Generally, simulation results were satisfying but simulated changes in topsoil pH need further verification with measurements.     

Old and New Cultivars of Soya Bean (Glycine max L.) Subjected to Soil Drying Differ in Abscisic Acid Accumulation,Water Relations Characteristics and Yield

Two old (Huangsedadou and Longxixiaohuangpi (LX)) and two new (Jindou 19 (JD) and Zhonghuang 30 (ZH)) soya bean (Glycine max (L.) Merr.) cultivars were used to investigate the influence of soil drying on the abscisic acid (ABA) accumulation in leaves, stomatal conductance (g_s), leaf water relations, osmotic adjustment (OA), leaf desiccation tolerance, yield and yield components. The greater ABA accumulation was induced by soil drying, which also inducing g_s decreased at higher soil water contents (SWC) and leaf relative water content (RWC) significantly decreased at lower SWC in the new soya bean cultivars than in the old soya bean cultivars. The soil water threshold between the value at which stomata began to close and the RWC began to decrease was significantly broader in the new cultivars than in the old cultivars. The new cultivars had significantly higher OA and lower lethal leaf water potential than old cultivars when the soil dried. The old cultivars had greater biomass, but lower grain yield than the new cultivars in well‐watered, moderate stress and severe stress conditions. Thus with soil drying, the new soya bean cultivars demonstrated greater adaptation to drought by inducing greater ABA accumulation, stomatal closure at higher SWC, enhanced OA and better water relations, associated with increased leaf desiccation tolerance, greater water use efficiency and higher yield.     

Effect of several insecticides on pomegranate aphid,Aphis punicae (Passerini) (Homoptera: Aphididae) and its predators under field conditions

The pomegranate aphid, Aphis punicae (Passerini) (Homoptera: Aphididae) is one of the most important pests in Egypt on pomegranate trees. The study presented in this paper was conducted from 2012 to 2013 to compare the efficiency and selectivity of neonicotinoids (acetamiprid, imidacloprid, thiamethoxam and dinotefuran), organophosphate (malathion) and carbamate (pirimicarb) insecticides against pomegranate aphid and the associated predators, Coccinella undecimpunctata Linnaeus, Chrysoperla carnea (Stephens) and Syrphus corolla Fabricius. These insecticides significantly reduced the pomegranate aphid population by an average of between 70.6 and 90.7%. The effects persisted for up to 15 days after the end of treatment. The greatest control of A. punicae was achieved by thiamethoxam > acetamiprid > imidacloprid > pirimicarb > dinotefuran > malathion. Acetamiprid, imidacloprid, thiamethoxam, malathion and pirimicarb were harmful to predators and the observed reduction ranged from 75.4 to 100% and dinotefuran was moderately harmful to lacewings with an average reduction about 64.4%. These results suggested that the neonicotinoid insecticides can be considered as promising candidates for controlling the pomegranate aphid as, although they do have harmful to moderately harmful effects on predators, they are significantly less toxic to these beneficial organisms than malathion and pirimicarb.     

Long‐Term Field Drought Affects Leaf Protein Pattern and Chloroplast Ultrastructure of Winter Wheat in a Cultivar‐Specific Manner

Recurrent drought periods of varying duration often cause extensive crop damage and affect wheat production in Southern Europe. This study compares biochemical and ultrastructural responses of four wheat (Triticum aestivum L.) cultivars to long‐term field drought, and their contribution to final grain yield. Gel electrophoresis and immunoblotting analyses combined with transmission electron microscopy and grain yield evaluation were employed to assess drought susceptibility of the wheat cultivars. Two of them behaved as drought‐tolerant, the other two presented as drought sensitive. Enhanced degradation of Rubisco large subunit (RLS), Rubisco small subunit (RSS) and Rubisco activase (RA) accompanied by an increased protease activity and reduced levels of heat shock proteins (HSP70) and dehydrins (DHNs) were associated with drought sensitivity. Drought tolerance coincided with relatively stable or increased HSP70 and DHN contents, and unchanged/higher levels of RLS, RSS and RA. Sensitive cultivars were more vulnerable to ultrastructural damages, showing obvious degradation of chloroplast membrane systems and depletion of leaf starch reserves. These drought responses affected yield potential, as tolerant cultivars gave higher yield under intense drought. Thus, our results provide additional insights into the complexity of plant drought responses, identifying multiple interacting traits that may serve as indirect selection criteria for wheat drought tolerance.     

Genetic relationships and hybrid vigour in olive (Olea europaea L.) by microsatellites

The olive (Olea europaea) is one of the most important oleaginous crops of the Mediterranean basin. Increased demand for olive oil creates a need for new olive varieties to help meet the requirements of the global market. However, olive breeding has been handicapped by such varied challenges as a prolonged juvenile period, agrotechnical problems and insufficient genetic knowledge. The use of DNA markers has the potential to overcome these problems and increase the effectiveness of classical breeding programmes. In this study, co‐dominant polymorphic simple sequence repeats (SSRs) were used as markers to analyse the genetic relationships between several local and other ‘non‐native’ olive cultivars. Cluster analysis revealed four major groups among the 15 cultivars examined in this study. Table and oil cultivars were clustered in different groups. However, the clusters did not differentiate between cultivars of different geographical origins. In addition, we used the data gathered to analyse genetic relationships to evaluate the effects of heterosis in agricultural traits. Genetic distances between cultivars were determined based on the SSR genotype data and were used for evaluating the possible effects of heterosis in various F₁ populations. Interestingly, phenotypic data of F₁ progenies from crosses between different cultivars indicated the potential effects of heterosis as expressed in several traits. Genetic distance between parents was significantly correlated to F₁ performance for three traits: percentage of dry fruit weight, oil content and commercial oil production. Thus, crosses between olive cultivars exhibiting relatively extensive genetic distances one from the other are expected to result in better progeny performance in future Olea breeding programmes. Our study linked assessment of biodiversity of commercial olive cultivars with the application of this information in olive breeding programmes for selection of specific parents to generate superior new cultivars.     

Expression of recombinant phytase of Bacillus subtilis E20 in Escherichia coli HMS 174 and improving the growth performance of white shrimp,Litopenaeus vannamei,juveniles by using phytase‐pretreated soybean meal‐containing diet

To produce a thermostable and neutral phytase (phy) of Bacillus subtilis E20 in Escherichia coli HMS174 and evaluate its efficiency in improving growth performance. The phy C of B. subtilis E20 was expressed in E. coli HMS 174, and then the 42‐kDa recombinant phy C was purified by Ni‐NAT and analysed by SDS–PAGE. The recombinant phy C had optimal ranges of pH of 6 ~ 7 and temperature of 50 ~ 60 °C. A thermostability analysis showed that the enzyme is a thermostable phytase, and around 33% of residual activity was detected after being incubated at 90 ~ 100 °C for 10 min. The recombinant phy C‐pretreated soybean meal for feed preparation improved white shrimp, Litopenaeus vannamei, growth and feed efficiency. Overall, the neutral and thermostable phy C is suitable for aquafeed, and it is able to improve the nutritional utilization, resulting in enhanced shrimp growth and reduced feed costs.     

Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.