Analyses of albumins, globulins and amphiphilic proteins by proteomic approach give new insights on waxy wheat starch metabolism

Starch is composed of two types of glucose polymers: amylose and amylopectin. The Waxy (Wx) locus controls amylose synthesis in the wheat kernel. Hexaploid wheat has three Wx loci located on chromosomes 7A (Wx-A1), 4A (Wx-B1), and 7D (Wx-D1). Eight near isogenic lines (NILs) of Triticum aestivum cv. Tremie with one, two or three Wx null alleles were used. The albumin–globulin fraction, and amphiphilic proteins were separated using 2-dimensional electrophoresis (2DE) allowing the changes in the waxy kernel to be identified. Albumin–globulin fraction showed overexpression of sucrose synthases in the waxy NILs compared to the normal form of Tremie and a decrease in many proteins related to stress and defence metabolism such as serpins. A subunit of ADP-glucose pyrophosphorylase (AGPase), which is known to play a major role in starch synthesis, was also shown to be down regulated in the waxy NILs. Amphiphilic proteins confirmed the observations made on the albumin–globulin fraction with a decrease in a stress-related protein. These different regulations linked to observations made on wheat kernel (thousand kernel weight (TKW), protein amount per grain, size and distribution of the starch granules) led to formulation of the hypothesis that waxy endosperm does not reach maturity of the wild-type endosperm.     

Impact of Distillery Effluent on Soil Carbon and Nitrogen Dynamics in a Vertisol of Central India

Distilleries produce a huge quantity of effluents, popularly known as spent wash (SW), which when bio-methanated produce post-methanation effluents (PME). A field experiment on soybean–wheat system was conducted for five consecutive years in a Vertisol of central India to evaluate the effect of distillery effluent (DE) on soil carbon and nitrogen dynamics. Ten treatment combinations consisting of control, 100% NPK + Farmyard Manure (FYM), and graded level of SW and PME were applied. Total carbon content of soil increased significantly with applications of FYM and DE. SW was found superior in enhancing carbon content of soil in comparison to PME. Farmyard Manure contributed more carbon toward the recalcitrant pool, whereas DE contributed more carbon toward the active and slow pool. Nitrogen (N) availability was significantly improved with the application of DE. Balanced application of DE may act as amendment for increasing C and N stocks in Vertisol.     

Organogenesis and foetal haemodynamics during the normal gestation of healthy black‐rumped agoutis (Dasyprocta prymnolopha,Wagler, 1831) bred in captivity

The objective of this study was to define the patterns of organogenesis and foetal haemodynamics during the normal gestation of healthy agoutis (Dasyprocta prymnolopha) kept in captivity. Thirty pregnant agoutis that ranged in size from small to medium and weighed between 2.5 and 3 kg underwent B‐mode and Doppler ultrasonography for the biometric evaluation of the foetal organs. The foetal aortic blood flow proved to be predominantly systolic, and the measured flow velocity was 78.89 ± 2.95 cm/s, with a maximum pressure gradient of 2.12 ± 0.27 mmHg. The liver was characterized by its large volume, occupying the entire cranial aspect of the abdominal cavity, and it was associated cranially with the diaphragm and caudally with the stomach. The flow velocity in the portal vein was estimated to equal 12.17 ± 2.37 cm/s, with a resistivity index of 0.82 ± 0.05. The gallbladder was centrally located and protruded cranially towards the diaphragm. The spleen was visualized as an elongated structure with tapered cranial and caudal extremities, and the foetal kidneys were visualized bilaterally in the retroperitoneal region, with the right kidney positioned slightly more cranially than the left. The morphological characterization and hemodynamic analysis of the foetal organs of black‐rumped agoutis via B‐mode and Doppler ultrasonography allow determination of the vascular network and of reference values for the blood flow required for perfusing the anatomical elements essential for maintaining the viability of foetuses at different gestational ages.     

Determinants of fenhexamid effectiveness against grey mould on grapevine: Respective role of spray timing,fungicide resistance and plant defences

Botrytis bunch rot of grapes is mainly controlled by applying fungicides at three crop growth stages: the end of flowering (BBCH 68), bunch closure (BBCH 77) and the beginning of veraison (BBCH 81). The hydroxyanilide derivative fenhexamid is among the most effective fungicides registered to control Botrytis cinerea. Its effectiveness was examined in relation to spray timing, fungicide resistance and defense responses of grapevine. Overall, the earlier fenhexamid was applied, the more effective it was at controlling B. cinerea. Frequencies of B. cinerea strains which were resistant to fungicides were evaluated at harvest. The frequencies of resistant phenotypes were similar among treatments and years with the exception of a class of multidrug resistant strain (MDR 2) whose frequency appeared to increase after fenhexamid applications. If current spray programs including fenhexamid appear to control bunch rot at the current MDR frequency, a propagation of MDR 2 strains might lead to a decline in disease control. Finally, defense responses were studied in grapevine flowers/berries following fenhexamid application. None of the defense processes tested was induced in flowers/berries at stages 68 and 77. Only an increase in chitinase activity was observed in treated-berries at stage 81, suggesting that fenhexamid effectiveness was not related to a stimulation of defense responses.     

Root phenotyping: important and minimum information required for root modeling in crop plants

As plants cannot relocate, they require effective root systems for water and nutrient uptake. Root development plasticity enables plants to adapt to different environmental conditions. Research on improvements in crop root systems is limited in comparison with that in shoots as the former are difficult to image. Breeding more effective root systems is proposed as the “second green revolution”. There are several recent publications on root system architecture (RSA), but the methods used to analyze the RSA have not been standardized. Here, we introduce traditional and current root-imaging methods and discuss root structure phenotyping. Some important root structures have not been standardized as roots are easily affected by rhizosphere conditions and exhibit greater plasticity than shoots; moreover, root morphology significantly varies even in the same genotype. For these reasons, it is difficult to define the ideal root systems for breeding. In this review, we introduce several types of software to analyze roots and identify important root parameters by modeling to simplify the root system characterization. These parameters can be extracted from photographs captured in the field. This modeling approach is applicable to various legacy root data stored in old or unpublished formats. Standardization of RSA data could help estimate root ideotypes.     

Motorways and bird traffic casualties: Carcasses surveys and scavenging bias

Most survey methods developed to estimate abundance of killed animals on motorways may be biased due to the unequal detectability of carcasses, their persistence time on the lanes, and scavengers activities. Unbiased surveys are needed to evaluate the relationships between bird casualties (mortality), motorways characteristics, and the neighbouring avifauna. The present study conducted on four motorways in France, aimed to evaluate factors affecting persistence and encounter probabilities and variations in scavenging activity to obtain unbiased estimates of bird traffic casualties. Each motorway was surveyed once per season during multiple years and we used capture–recapture methods to estimate detection and carcass persistence rates. Results showed that surveys by car were as efficient as surveys by foot in detecting carcasses on the pavement, but less efficient for carcasses on verges. Passeriformes represented the most numerous casualties, and the Barn Owl (Tyto alba) was the most frequently killed species. Encounter probabilities were constant and high (0.957 ± 0.007). Average daily persistence probability was 0.976 ± 0.003. Persistence probabilities were higher for large and old carcasses, during summer, and differed between seasons, but were relatively similar between years. Scavenging activities, estimated using experimental carcasses disposed on the safe lanes of motorways, varied between years, seasons, and differed between diurnal and nocturnal periods. A peak in scavenging activity occurred during diurnal periods in spring. Results suggest that surveys must take into account carcass characteristics and seasonal variability to obtain unbiased estimates of road killed birds on motorways, as well as variation in scavenging rates.     

Influence of EDTA and citric acid on lead-induced oxidative stress to Vicia faba roots

Purpose

In spite of substantial advancement in recent past, the role of metal speciation in assessing biogeochemical behaviour of Pb is still topical. Organic ligands are capable to modify Pb speciation in nutrient/soil solution and in turn its soil–plant transfer and toxicity. In this sense, the main objective of this study was to evaluate the effect of organic ligands on Pb-induced oxidative stress to Vicia faba roots.

Materials and methods

V. faba seedlings grown to controlled hydroponic system were treated with 5 μM Pb as lead nitrate in the presence and absence of organic ligands viz ethylenediaminetetraacetic acid (EDTA) and citric acid (CA) for 1, 4, 8, 12, and 24 h. The chemical speciation of Pb (percent free and chelated Pb) in nutrient solution in the presence and absence of organic ligands was calculated using Visual Minteq speciation model. The effect of chemical speciation on Pb-induced oxidative stress to V. faba roots was investigated using plant enzymatic antioxidative system [superoxide dismutases (SOD), guaiacol peroxidise (GPX), ascorbate peroxidase (APX), glutathione reductase (GR), and catalase (CAT)]. The antioxidant enzymes activities were determined using ultraviolet spectrophotometer.

Results and discussion

The activities of SOD, GPX, APX, and GR significantly increased whereas that of CAT decreased in V. faba roots under Pb alone treatment. Lead-induced increase/decrease in antioxidant enzymes activities was not linear but varies with treatment exposure time. EDTA dose dependently inhibited Pb-induced changes in antioxidant enzymes activities. However, CA did not cause any significant change in Pb-induced variation in antioxidant enzymes activities, but delayed or slightly reduced the Pb effect.

Conclusions

The present study suggested that physiological responses of V. faba roots to Pb toxicity vary with applied Pb form and duration of exposure. EDTA can inhibit Pb-induced toxicity to V. faba seedlings by forming stable Pb-EDTA complexes due to its high binding strength for Pb. However, CA had no effect on Pb-induced toxicity to V. faba roots due to weak complexation with Pb.     

Differences between the ovine tonsils based on an immunohistochemical quantification of the lymphocyte subpopulations

In sheep, the pharyngeal first defence line against oral and inhaled antigens is organized in six tonsils. Since tonsils are regarded as secondary lymphoid tissue and part of the acquired immune system which is subjected to induction through contact with antigens, an evaluation of the different lymphocyte populations in tonsils is useful to determine a tendency of the specific tonsils to more inductive or more effective immunity. By means of immunohistochemistry, different lymphocyte populations were quantified and localized using a panel of eight antibodies, i.e. anti-CD45, anti-CD21, anti-CD2, anti-CD3, anti-CD4, anti-CD8, anti-WC1 and anti-Ki67. The CD21+ B lymphocytes were localized within the tonsillar lymphoid follicles. The CD2+/CD3+ T lymphocytes were numerous in the interfollicular regions and were aligned underneath and within the epithelium but were also observed at the CD21+ pole of the lymphoid follicles. Near the lingual and tubal tonsils, and the tonsil of the soft palate, the CD45+ cells around the seromucous glands and in the lamina propria were mainly CD3+ T cells. In all tonsils, the WC1+ gamma delta T cells formed a small lymphocyte population which harboured the lamina propria and the interfollicular region. The relative percentages of the different lymphocyte populations of the large palatine and pharyngeal tonsils, which are macroscopically the most developed, were comparable. In contrast, the lingual tonsil was significantly different from the other tonsils not only by its small size and lack of lymphoid follicles, but also by the lymphocyte populations. Based on the lymphocyte populations, the ovine tonsils can be divided in three groups with the tonsil of the soft palate, the tubal and paraepiglottic tonsil forming an intermediate between the palatine and pharyngeal tonsils as true tonsils on the one side, and the lingual tonsil as a scattered lymphocyte aggregation on the other side.