Pectate lyase-like lubricates the male gametophyte's path toward its mating partner.

The pollen tube is an extension of the male gametophyte in plants and mediates sexual reproduction by delivering the sperm cells to the female gametophyte. To accomplish this task, the elongating pollen tube must break through the thick wall of the pollen grain and penetrate multiple pistillar tissues. Both processes require the loosening of cell wall material-that of the pollen intine and that of the apoplast of the transmitting tract. The enzymatic toolbox for these cell wall modifying processes employed by the invading male gametophyte is elusive. We investigated the role of the pectin-digesting pectate lyase-like (PLL) by combining mutant analysis with microscopy observations, fluorescence recovery after photo-bleaching experiments, and immuno-detection. We show that in Arabidopsis (Arabidopsis thaliana), PLLs are required for intine loosening during the first steps of pollen tube germination. We provide evidence that during pollen tube elongation, PLLs are released by the pollen tube into the extracellular space, suggesting that they may be employed to soften the apoplast of the transmitting tissue. The synergistic enzymatic action of PLLs in the pollen grain, the pollen tube, and the transmitting track contribute to an effective fertilization process.

Effects of Origanum majorana essential oil and antibiotics on the quality of frozen thawed Beni Arouss buck semen.

This study aims to investigate the effects of Origanum majorana (OM) essential oil (EO) at different concentrations and antibiotics on post-thawed Beni Arouss buck semen quality. Semen collection was performed using artificial vagina from eight Beni Arouss bucks. Ejaculates were pooled, divided into 12 equal aliquots, washed and diluted to 400 × 106 sperm/ml (with 7% of glycerol). Skim milk-based extender was supplemented with different concentrations of OM EO (0%; 0.01%; 0.02%; 0.03%; 0.04% and 0.05%) without antibiotics, marked as (CTR-), (M1-), (M2-), (M3-), (M4-) and (M5-), and with 50 mg of streptomycin and 50,000 IU of penicillin per 100 ml, marked as (CTR+), (M1+), (M2+), (M3+), (M4+) and (M5+), respectively. Aliquots were cooled to 4°C, then frozen in 0.25 ml straws with a programmable freezer and finally stored in liquid nitrogen for 48 h. Thawing was performed at 37°C for 30 s. Motility, live sperm, sperm abnormalities, membrane integrity, lipid peroxidation and bacterial growth were evaluated after thawing. Among the tested extenders, M2+ improved all semen quality parameters. Sperm motility, live sperm and membrane integrity increased significantly, while the number of abnormal sperm and bacterial growth decreased significantly. The toxic effect of OM EO, with and without antibiotics, appeared beyond 0.03%. In conclusion, M2+ is recommended to improve the cryopreservation of Beni Arouss buck semen.

Mechanosensitive ion channels contribute to mechanically evoked rapid leaflet movement in Mimosa pudica.

Mechanoperception, the ability to perceive and respond to mechanical stimuli, is a common and fundamental property of all forms of life. Vascular plants such as Mimosa pudica use this function to protect themselves against herbivory. The mechanical stimulus caused by a landing insect triggers a rapid closing of the leaflets that drives the potential pest away. While this thigmonastic movement is caused by ion fluxes accompanied by a rapid change of volume in the pulvini, the mechanism responsible for the detection of the mechanical stimulus remains poorly understood. Here, we examined the role of mechanosensitive ion channels in the first step of this evolutionarily conserved defense mechanism: the mechanically evoked closing of the leaflet. Our results demonstrate that the key site of mechanosensation in the Mimosa leaflets is the pulvinule, which expresses a stretch-activated chloride-permeable mechanosensitive ion channel. Blocking these channels partially prevents the closure of the leaflets following mechanical stimulation. These results demonstrate a direct relation between the activity of mechanosensitive ion channels and a central defense mechanism of M. pudica.

Estimation of Grazing Activity of Dairy Goats Using Accelerometers and Global Positioning System.

The recent development of advanced electronic sensors to monitor and record animal grazing activity provides a real opportunity to facilitate understanding of their behavioral responses. This study aimed to characterize the grazing activity and protein−energy requirements of grazing dairy goats in a Mediterranean woodland in northern Morocco by combining two sensors, the Global Positioning System (GPS) and three axis accelerometers. An experiment was conducted in a representative woodland with eight dairy goats. Measurements were undertaken during the three main grazing seasons (spring, summer, and autumn) for two consecutive and contrasting years. Grazing activity parameters were assessed using GPS collars and leg position sensors. The results showed that grazing time was higher in spring (57% and 59%) than in summer (39% and 36%) and autumn (41% and 45%), respectively, for the dry and wet years (p < 0.001). During the two studied years, the daily horizontal distance traveled by the goats increased from spring (about 4.5 km) to summer (about 6.5 km) and autumn (about 7.4 km), while greater daily vertical distances were recorded over similar distances during summer−autumn. Several protein−energy intakes that were insufficient to cover the requirements of the grazing goats were recorded, especially in summer. The combination of GPS collars and accelerometers contributed to a better understanding of the grazing activities of dairy goats in the studied Mediterranean woodland. These findings provide useful data on the protein−energy balances of dairy goats and offer additional information that could be useful for herders and managers to enhance goat-feeding strategies and guarantee high-performance in the semi-extensive traditional goat farming system.

Using GPS Collars and Sensors to Investigate the Grazing Behavior and Energy Balance of Goats Browsing in a Mediterranean Forest Rangeland.

The Global Positioning System (GPS) and sensors technologies are increasingly used to study the grazing behavior of animals. This work was conducted to understand the grazing behavior and energy balance of goats browsing in forest rangeland using GPS and sensors technologies. Forage availability was estimated using the quadrat method during three grazing seasons. Simultaneously, eight indigenous goats were selected to explore their feeding behavior, grazing activities, and energy requirements. The experimental goats were fitted with GPS collars and leg sensors to monitor their grazing activities. At the same time, direct observation was used as a method to study their feeding behavior. Forage availability was higher during spring compared to the summer and autumn seasons. Goats recorded the highest biting rate during summer and autumn (about 22 bites/min). The highest intake rate was recorded during spring (5.6 g DM/min). During spring, goats spent most of their time on grazing (48%) in contrast to the summer and autumn (<31%; p < 0.001). They prolonged their lying down time in summer at the expense of standing duration. The time devoted exclusively to grazing (eating) was longer in spring. Walking time in summer and autumn was longer than in spring (p < 0.001). During summer and autumn, the energy balance of goats under grazing conditions was in deficit. Using GPS collars and leg sensors appears to be a useful and easily replicable method to explore and understand the seasonal changes in the grazing areas and activities of goats in a mountainous region. The results could help goat herders and managers to develop feeding and grazing systems while increasing the performance of goats in the Mediterranean forest rangeland.

Pectin Chemistry and Cellulose Crystallinity Govern Pavement Cell Morphogenesis in a Multi-Step Mechanism.

Simple plant cell morphologies, such as cylindrical shoot cells, are determined by the extensibility pattern of the primary cell wall, which is thought to be largely dominated by cellulose microfibrils, but the mechanism leading to more complex shapes, such as the interdigitated patterns in the epidermis of many eudicotyledon leaves, is much less well understood. Details about the manner in which cell wall polymers at the periclinal wall regulate the morphogenetic process in epidermal pavement cells and mechanistic information about the initial steps leading to the characteristic undulations in the cell borders are elusive. Here, we used genetics and recently developed cell mechanical and imaging methods to study the impact of the spatio-temporal dynamics of cellulose and homogalacturonan pectin distribution during lobe formation in the epidermal pavement cells of Arabidopsis (Arabidopsis thaliana) cotyledons. We show that nonuniform distribution of cellulose microfibrils and demethylated pectin coincides with spatial differences in cell wall stiffness but may intervene at different developmental stages. We also show that lobe period can be reduced when demethyl-esterification of pectins increases under conditions of reduced cellulose crystallinity. Our data suggest that lobe initiation involves a modulation of cell wall stiffness through local enrichment in demethylated pectin, whereas subsequent increase in lobe amplitude is mediated by the stress-induced deposition of aligned cellulose microfibrils. Our results reveal a key role of noncellulosic polymers in the biomechanical regulation of cell morphogenesis.

Plant AP180 N-Terminal Homolog Proteins Are Involved in Clathrin-Dependent Endocytosis during Pollen Tube Growth in Arabidopsis thaliana.

Polarized cell growth in plants is maintained under the strict control and exquisitely choreographed balance of exocytic and endocytic membrane trafficking. The pollen tube has become a model system for rapid polar growth in which delivery of cell wall material and membrane recycling are controlled by membrane trafficking. Endocytosis plays an important role that is poorly understood. The plant AP180 N-Terminal Homolog (ANTH) proteins are putative homologs of Epsin 1 that recruits clathrin to phosphatidylinositol 4, 5-bisphosphate (PIP2) containing membranes to facilitate vesicle budding during endocytosis. Two Arabidopsis ANTH encoded by the genes AtAP180 and AtECA2 are highly expressed in pollen tubes. Pollen tubes from T-DNA inserted knockout mutant lines display significant morphological defects and unique pectin deposition. Fluorescent tagging reveals organization into dynamic foci located at the lateral flanks of the pollen tube. This precisely defined subapical domain coincides which clathrin-mediated endocytosis (CME) and PIP2 localization. Using a liposome-protein binding test, we showed that AtECA2 protein and ANTH domain recombinant proteins have strong affinity to PIP2 and phosphatidic acid containing liposomes in vitro. Taken together these data suggest that Arabidopsis ANTH proteins may play an important role in CME, proper cell wall assembly and morphogenesis.

Application of Precision Technologies to Characterize Animal Behavior: A Review.

This study aims to evaluate the state of precision livestock farming (PLF)'s spread, utilization, effectiveness, and evolution over the years. PLF includes a plethora of tools, which can aid in a number of laborious and complex tasks. These tools are often used in the monitoring of different animals, with the objective to increase production and improve animal welfare. The most frequently monitored attributes tend to be behavior, welfare, and social interaction. This study focused on the application of three types of technology: wearable sensors, video observation, and smartphones. For the wearable devices, the focus was on accelerometers and global positioning systems. For the video observation, the study addressed drones and cameras. The animals monitored by these tools were the most common ruminants, which are cattle, sheep, and goats. This review involved 108 articles that were believed to be pertinent. Most of the studied papers were very accurate, for most tools, when utilized appropriate; some showed great benefits and potential.

Carvacrol and Thymol Enhance the Quality of Beni Arouss Buck Semen Stored at 4 °C Thanks to Their Antimicrobial Properties.

This study aims to investigate the impact of carvacrol and thymol on the quality of Beni Arouss buck semen stored in skim milk at 4 °C. Ejaculates were collected from eight Beni Arouss bucks weekly for 11 weeks, pooled, and then divided into three equal parts. Samples were diluted to 400 × 106 sperm/mL in skim milk (control) and skim milk supplemented with a single dose of 200 µM carvacrol and thymol each. Evaluations of sperm motility, viability, abnormalities, membrane integrity, lipid peroxidation, and bacterial growth were conducted at 0, 6, 24, and 48 h of liquid storage at 4 °C. After 48 h of storage, the results indicate that the addition of carvacrol positively influences total and progressive motility and viability. However, it also leads to a decrease in lipid peroxidation and bacterial growth compared to the control group (p < 0.05). Thymol showed similar results to carvacrol, except for progressive motility (p > 0.05). Bacterial growth was negatively correlated with total and progressive motility and viability (p < 0.05), while no correlation between lipid peroxidation and these parameters was observed (p > 0.05). In conclusion, the addition of carvacrol and thymol to skim milk extender moderately improves the quality of Beni Arouss buck semen after 48 h storage at 4 °C due to its antimicrobial activity.

The cell wall of the Arabidopsis pollen tube--spatial distribution, recycling, and network formation of polysaccharides.

The pollen tube is a cellular protuberance formed by the pollen grain, or male gametophyte, in flowering plants. Its principal metabolic activity is the synthesis and assembly of cell wall material, which must be precisely coordinated to sustain the characteristic rapid growth rate and to ensure geometrically correct and efficient cellular morphogenesis. Unlike other model species, the cell wall of the Arabidopsis (Arabidopsis thaliana) pollen tube has not been described in detail. We used immunohistochemistry and quantitative image analysis to provide a detailed profile of the spatial distribution of the major cell wall polymers composing the Arabidopsis pollen tube cell wall. Comparison with predictions made by a mechanical model for pollen tube growth revealed the importance of pectin deesterification in determining the cell diameter. Scanning electron microscopy demonstrated that cellulose microfibrils are oriented in near longitudinal orientation in the Arabidopsis pollen tube cell wall, consistent with a linear arrangement of cellulose synthase CESA6 in the plasma membrane. The cellulose label was also found inside cytoplasmic vesicles and might originate from an early activation of cellulose synthases prior to their insertion into the plasma membrane or from recycling of short cellulose polymers by endocytosis. A series of strategic enzymatic treatments also suggests that pectins, cellulose, and callose are highly cross linked to each other.

Finite element model of polar growth in pollen tubes.

Cellular protuberance formation in walled cells requires the local deformation of the wall and its polar expansion. In many cells, protuberance elongation proceeds by tip growth, a growth mechanism shared by pollen tubes, root hairs, and fungal hyphae. We established a biomechanical model of tip growth in walled cells using the finite element technique. We aimed to identify the requirements for spatial distribution of mechanical properties in the cell wall that would allow the generation of cellular shapes that agree with experimental observations. We based our structural model on the parameterized description of a tip-growing cell that allows the manipulation of cell size, shape, cell wall thickness, and local mechanical properties. The mechanical load was applied in the form of hydrostatic pressure. We used two validation methods to compare different simulations based on cellular shape and the displacement of surface markers. We compared the resulting optimal distribution of cell mechanical properties with the spatial distribution of biochemical cell wall components in pollen tubes and found remarkable agreement between the gradient in mechanical properties and the distribution of deesterified pectin. Use of the finite element method for the modeling of nonuniform growth events in walled cells opens future perspectives for its application to complex cellular morphogenesis in plants.

Live Fluorescence Visualization of Cellulose and Pectin in Plant Cell Walls.

The plant cell wall comprises various types of macromolecules whose abundance and spatial distribution change dynamically and are crucial for plant architecture. High-resolution live cell imaging of plant cell wall components is, therefore, a powerful tool for plant cell biology and plant developmental biology. To acquire suitable data, the experimental setup for staining and imaging of non-fixed samples must be straightforward and avoid creating stress-induced artifacts. We present a detailed sample preparation and live image acquisition protocol for fluorescence visualization of cell wall components using commercially available probes and stains.

Cytoskeletal regulation of primary plant cell wall assembly.

The plant cell wall is an extracellular matrix that envelopes cells, gives them structure and shape, constitutes the interface with symbionts, and defends plants against external biotic and abiotic stress factors. The assembly of this matrix is regulated and mediated by the cytoskeleton. Cytoskeletal elements define where new cell wall material is added and how fibrillar macromolecules are oriented in the wall. Inversely, the cytoskeleton is also key in the perception of mechanical cues generated by structural changes in the cell wall as well as the mediation of intracellular responses. We review the delivery processes of the cell wall precursors that are required for the cell wall assembly process and the structural continuity between the inside and the outside of the cell. We provide an overview of the different morphogenetic processes for which cell wall assembly is a crucial element and elaborate on relevant feedback mechanisms.

Effect of Olive Cake and Cactus Cladodes Incorporation in Goat Kids' Diet on the Rumen Microbial Community Profile and Meat Fatty Acid Composition.

The olive cake (OC) and the cactus cladodes (CC) are two alternative feed resources widely available in the southern Mediterranean region that could be used in ruminants' diet. Their impact on the rumen bacterial ecosystem is unknown. This work aims to evaluate their effects on the microbial community and meat fatty acids of goat's kids. Forty-four goat kids were divided into four groups receiving diets with conventional concentrate, or 35% OC, or 30% CC, or 15% OC, and 15% CC. After 3 months, these animals were slaughtered, and the rumen liquor and longissimus dorsi and semimembranosus muscles samples were collected. Animals receiving a control diet had rumen liquor with high acidity than test groups (p < 0.001). Test rumen liquor was more adapted to digest efficiently their matching diet than control liquor (p < 0.05). These feedstuffs did not affect rumen bacteria abundance and alpha diversity (richness, evenness, and reciprocal Simpson indexes), and these results were confirmed by beta-diversity tests (NMDS plot, HOMOVA, PERMANOVA). The test diets slightly affected the individual fatty acids of meat (p < 0.05) without effect on fatty acids summaries, indexes, and ratios. Thus, these alternative feed resources could take place in goat kids' diet to diversify their feed and to reduce feed costs.

Temporal Variations in Chemical Composition, In Vitro Digestibility, and Metabolizable Energy of Plant Species Browsed by Goats in Southern Mediterranean Forest Rangeland.

Forest rangelands contribute largely to goat diets in the Mediterranean area. Information about browsed plant quality is essential for adequate feeding management. The purpose of this study was to evaluate the temporal changes in chemical composition and in vitro digestibility of the main plant species selected by goats in the Southern Mediterranean forest rangeland during two consecutive years; these were very contrasted (dry and wet). The browsed species were composed of herbaceous, eleven shrubs, and four tree species. Overall, large variability in chemical composition, in vitro organic matter digestibility (IVOMD), and metabolizable energy (ME) was observed among species, grazing season (spring, summer, and autumn), and years within each species. Crude protein (CP) content varied from 60 to 240 g/kg dry matter (DM). The fiber fractions, except for Quercus suber, increased significantly by advancing maturity. Due to the water stress, the lignin level presented a higher value during the spring of the dry year. Condensed tannin (CT) content varied from 2 to 184 g/kg DM. CP, IVOMD, and ME showed a negative correlation with lignin and CT. Based on the results presented herein, it is concluded that the nutritive value of the browsed plant species was highest in the spring and lowest during the summer and autumn of both studied years. With a good grazing management strategy, the selected plant species by goats could guarantee high-quality feeding resources throughout the year.

Effect of Dried Mealworms (Tenebrio molitor), Larvae and Olive Leaves (Olea europaea L.) on Growth Performance, Carcass Yield and Some Blood Parameters of Japanese Quail (Coturnix coturnix japonica).

The aim of this study was to investigate the effect of Tenebrio molitor meal (TM) and/or olive leaf powder (OL) supplementation to quail diet on animal performance, carcass yield and some blood parameters. One hundred and forty-four 1-day-old Japanese quails (body weight: 29.9 ± 0.46 g) were divided into four groups of 36 chicks, receiving four different diets, i.e., G1: Standard commercial (SCD); G2: SCD + 3% TM; G3: SCD + 3% OL; and G4: SCD + 3% TM + 2% OL. Feed and water were provided ad-libitum. The results showed that TM and OL inclusion promoted quail body weight at 5 weeks of age; p = 0.001. Feed conversion ratio (FCR) of G3 was significantly (p < 0.01) reduced compared to the other groups. Overall mortality was not different, at around 6.25%, between groups. Carcass [(71.5-74.3%)], gizzard [(2.5-2.7%)], heart [(1.1-1.2%)] and giblet [(6.8-6.9%)] yields were not influenced by the diet. No significant effect of the diet was observed for serum proteins, creatinine, urea and lipids. To conclude, the mixture of an incorporation of TM and OL in quail diet showed no adverse effects on growth performance, carcass constituent yields and serum constituents.

Morphogenesis of complex plant cell shapes: the mechanical role of crystalline cellulose in growing pollen tubes.

Cellulose is the principal component of the load-bearing system in primary plant cell walls. The great resistance to tensile forces of this polysaccharide and its embedding in matrix components make the cell wall a material similar to a fiber composite. In the rapidly growing pollen tube, the amount of cellulose in the cell wall is untypically low. Therefore, we want to investigate whether the load-bearing function of cellulose is nevertheless important for the architecture of this cell. Enzymatic digestion with cellulase and inhibition of cellulose crystal formation with CGA (1-cyclohexyl-5-(2,3,4,5,6-pentafluorophenoxy)-1lambda4,2,4,6-thiatriazin-3-amine) resulted in the formation of tubes with increased diameter in Solanum chacoense and Lilium orientalis when present during germination. In pre-germinated tubes, application of both agents resulted in the transient arrest of growth accompanied by the formation of an apical swelling indicating a role in the mechanical stabilization of this cellular region. Once growth resumed in the presence of cellulase, however, the cell wall in the newly formed tube showed increased amounts of pectins, possibly to compensate for the reduced amount of cellulose. Scanning electron microscopy of pollen tubes subjected to digestion of matrix polysaccharides revealed the mechanical anisotropy of the cell wall. In both Lilium and Solanum, the angle of highest stability revealed by crack formation was significantly below 45 degrees , an indication that in the mature part of the cell cellulose may not the main stress-bearing component against turgor pressure induced tensile stress in circumferential direction.

Silicone Chambers for Pollen Tube Imaging in Microstructured In Vitro Environments.

Live cell imaging at high resolution of pollen tubes growing in vitro requires an experimental setup that maintains the elongated cells in a single optical plane and allows for controlled exchange of growth medium. As a low-cost alternative to lithography-based microfluidics, we developed a silicone-based spacer system that allows introducing spatial features and flexible design. These growth chambers can be cleaned and reused repeatedly.

Foraging Behavior of Goats Browsing in Southern Mediterranean Forest Rangeland.

Mediterranean forest rangelands offer an important feed source for goats. Concerns about grazing strategies and management schemes in order to ensure the rangeland sustainability of Southern Mediterranean forest have revived interest in the foraging behavior of goats. This study was conducted to investigate the seasonal changes of feeding behavior of grazing goats in the Southern Mediterranean forest rangeland of Northern Morocco during two consecutive years beginning in 2016. The direct observation method was used to compare diet composition, intake rate, and diet selectivity of goats during three seasons (spring, summer, and fall). Bite mass of each plant species selected by goats was estimated using hand-plucked simulation. The optimal foraging theory was used as a tool to explain the goats foraging decisions. Bite mass range was extremely wide and varied seasonally. The goats' diet was largely composed of Cistus spp., Lavandula stoechas, Quercus spp., and Myrtus communis. The result shows that the smaller the bite mass, the higher the biting rate, leading to increased short term intake rates. The selection of various plant species during fall and summer enlarged the diet diversity of goats. As expected, goats preferred trees and some shrubs despite their low availability. Consequently, the most available species is not necessarily the most positively selected. Particular high and positive selection of Quercus suber was observed over seasons. The outcomes confirm the high adaptability and ability of goats to select a woody species across seasons. Knowledge about forage availability and the feeding behavior of goats could be used as the first guide for rangeland managers to ensure herd and forest sustainability.

Optimization of conditions for germination of cold-stored Arabidopsis thaliana pollen.

One of the rare weak points of the model plant Arabidopsis is the technical problem associated with the germination of its male gametophyte and the generation of the pollen tube in vitro. Arabidopsis pollen being tricellular has a notoriously low in vitro germination compared to species with bicellular pollen. This drawback strongly affects the reproducibility of experiments based on this cellular system. Together with the fact that pollen collection from this species is tedious, these are obstacles for the standard use of Arabidopsis pollen for experiments that require high numbers of pollen tubes and for which the percentage of germination needs to be highly reproducible. The possibility of freeze-storing pollen after bulk collection is a potential way to solve these problems, but necessitates methods that ensure continued viability and reproducible capacity to germinate. Our objective was the optimization of germination conditions for Arabidopsis pollen that had been freeze-stored. We optimized the concentrations of various media components conventionally used for in vitro pollen germination. We found that in general 4 mM calcium, 1.62 mM boric acid, 1 mM potassium, 1 mM magnesium, 18% sucrose at pH 7 and a temperature of 22.5 degrees C are required for optimal pollen germination. However, different experimental setups may deviate in their requirements from this general protocol. We suggest how to optimally use these optimized methods for different practical experiments ranging from morphological observations of pollen tubes in optical and electron microscopy to their bulk use for molecular and biochemical analyses or for experimental setups for which a specific medium stiffness is critical.