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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.

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.

The aim of this study was to evaluate the feeding behaviour of indigenous goats and the selection drivers of different plant species in a Mediterranean forest rangeland. To achieve this goal, the seasonal variations in terms of forage availability and quality of ingested plant species were studied during three grazing seasons. In the same period, eight indigenous goats of Beni Arouss breed were selected to explore the seasonal changes in their browsing behaviour. Forage quality was determined by the hand-plucking technique. The results showed a wide seasonal variation in forage availability and quality, and feeding behaviour. Woody species were more selected independently of the season (p < 0.001). The crude protein content varied from 53.3 g/kg of dry mater (DM) for Erica arborea in summer to 197 g/kg DM for Calicotome villosa in autumn (p < 0.001). Despite the high condensed tannins content in selected shrubs, they were highly consumed. In vitro organic matter digestibility (IVOMD) and metabolizable energy (ME) had recorded the highest contents in herbaceous during spring. Shrubs and trees contain the lowest levels of IVOMD (

In the Mediterranean area, the olive cake (OC) and cactus cladodes (CC) are two alternative resources widely available that could be used for ruminants’ feeding. The objective of this study was to evaluate the effects of OC and/or CC diet incorporation on the production performance and quality of goat milk. Forty-four lactating goats were randomly allocated to four groups. The control one (Co) received a conventional feed. Test groups (TOC; TCC and TOC+CC) received 20% OC, 30% CC, or 15% OC and 20% CC, respectively, on concentrate dry matter basis. Over three months, milk production was evaluated, and samples were collected to analyze the milk quality. No significant differences were observed between control and test groups for daily milk production, yield, composition and acidity. In milk fat, OC incorporation increased C18:1n-9, mono-unsaturated (MUFA) and n-9 fatty acid (FA), and decreased 9t-C18:1 and poly-unsaturated FA (PUFA) (p < 0.05). Significantly highest contents of C15:0, C18:1n-9, and C21:0, and lowest levels of C4:0, 9t-C18:1, 6t-C18:2, C20:0, and PUFA were obtained with cactus cladodes administration (p < 0.05). The TOC + CC diet reduced C4:0, 9t-C18:1, 6t-C18:2, C22:6n-3, and PUFA proportions, and increased C18:1n-9, MUFA/PUFA, and thrombogenic indexes. The incorporation rates of OC and CC that could reach 20% and 30%, respectively, had no major negative effects on milk production performance, composition, and quality. Thus, they could be introduced in the diets of lactating goats.

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.

In the Mediterranean basin, the silvopastoral system has always been part of the regional economy. Silvopastoral areas contribute significantly to the development of the ecological and socio-economic factors of territorial sustainability. However, little attention has been paid to reviewing the importance, historical evolution, socio-economic conditions, and climate change impact of the southern Mediterranean region of northern Morocco. This review casts light on research on the silvopastoral systems in Morocco with a focus on the Mediterranean side. Sixty-nine peer-reviewed publications were selected for this systematic review. The findings revealed that silvopastoral areas play a very important role in animal feeding in Morocco. Animals are managed traditionally on these woodlands where they graze throughout the year. The silvopastoral system has undergone several changes over recent decades. These changes have had a direct impact on ecosystems, causing a decline in biodiversity, forest degradation, and an increase in land use at the expense of silvopastoral areas, mainly in the Mediterranean region of northern Morocco. Consequently, the sustainability of the silvopastoral systems would undoubtedly be negatively affected. The review finishes with some crucial strategies and propositions that could be encouraged to support sustainable management actions of silvopastoral resources. In the conclusion, we outline the need to assess the realistic socio-economic and ecological benefits of silvopastoral systems to promote their sustainable development.

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., One-sentence summary: Vascular plants use mechanosensitive ion channels to move their leaflets in response to external mechanical stimuli.

Summary 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.

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.

Background The current study was carried out to evaluate the effect of olive cake and cactus cladodes incorporation on carcass characteristics and meat quality of goat kids.Methods Forty-eight male goat kids were divided into four groups. The control group received a conventional supplementation and the test groups were supplemented with a concentrate containing, on dry matter basis, 35% of olive cake (OC), 30% of cactus cladodes (CC), or 15% OC and 15% CC (OC+CC) respectively. After 3 months, all animals were slaughtered and the carcass quality was characterized. Samples of Longissimus dorsi and Semimembranosus were collected to determine meat quality.Results The diets did not affect final body weight and carcass characteristics, except for muscle index that decreases with 35% OC, and redness and yellowness at tail outline and belly (P < 0.05). The OC incorporation increased yellowness at tail and decreased redness at belly (P < 0.05), while the CC inclusion decreased redness at tail outline (P < 0.01), and redness (P < 0.05) and yellowness at belly (P < 0.01). Meat ultimate pH, color, moisture and tenderness were not affected by diets. In Longissimus dorsi, higher proteins (P < 0.001) and lower fat and ash (P < 0.05) content were observed with CC, and lower proteins content with OC and OC+CC (P < 0.001). In Semimembranosus, a low initial pH (P < 0.05) was observed with OC and high protein content (P < 0.001) with CC and OC+CC. Generally, groups, ratios, and indexes of fatty acids (FA) were similar between groups, except FA profile that was affected by diet, especially for Semimembranosus. In Longissimus dorsi, the OC and CC introduction decreased C16:1, and C20: 3n3 increased with CC and OC+CC (P < 0.05). While in Semimembranosus, OC increased C6, C8, C18:3n3, C20:2 and C22:2, whereas CC incorporation reduced C8, C10, and C15:1, and OC+CC reduced C10 (P < 0.05).Conclusions This study was shown that OC and CC could be introduced in goat kids’ diet, without a negative effect on carcass characteristics and meat quality.

Plant cell walls constitute the extracellular matrix surrounding plant cells and are composed mainly of polysaccharides. The chemical makeup of the primary plant cell wall, and specifically, the abundance, localization and arrangement of the constituting polysaccharides are intimately linked with growth, morphogenesis and differentiation in plant cells. Visualization of the cell wall components is, therefore, a crucial tool in plant cell developmental studies. In this technical update, we present protocols for fluorescence visualization of cellulose and pectin in selected plant tissues and illustrate examples of some of the available labels that hold promise for live imaging of plant cell wall expansion and morphogenesis.

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.

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&rsquo, 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.

In northern Morocco, forest and silvopastoral areas have always formed an integral part of goat feeding and consequently of extensive livestock systems. However, it was reported that these areas are now going through a gradual degradation mainly due to overgrazing, resulting in land use-land cover (LULC) changes. Doubts persist about actual goat grazing impacts on forest and silvopastoral cover changes. Detecting and understanding drivers of LULC changes has become a central component of current strategies for managing natural resources to implement appropriate development policies. The objectives of this study were to assess the forest and silvopastoral cover changes and transition among five major land cover classes that took place in the last three decades (1984–2014), to verify the hypothesis that goats act as a main driver of forest and silvopastoral degradation, and to explore the perception of stakeholders based on remote sensing data, field surveys, and overall statistics. Between 1984 and 2014, forest and silvopastoral areas declined by 25% (matorral included). Farmers and local institutions all agreed that drought, fire, soil erosion, and population growth are the most striking drivers of forest and silvopastoral decreases. One being utilitarian and the other protectionist, they have conflicting perceptions about the effect of agricultural expansion, authority policy, deforestation, and overgrazing. The forest cover change is impacted by interaction among different change drivers. The levels of forest degradation depend on the intensity of this interaction. The outcomes confirm the alarming decrease in the amount of forest and silvopastoral cover. These results can be used as the first guide for future studies and decision-makers, focusing on the real drivers of forest and silvopastoral degradation, so as to develop targeted intervention to secure sustainable and sufficient resources needed for animal and population well-being.

Cell and organ morphogenesis in plants are regulated by the chemical structure and mechanical properties of the extracellular matrix, the cell wall. The two primary load bearing components in the plant cell wall, the pectin matrix and the cellulose/xyloglucan network, are constantly remodelled to generate the morphological changes required during plant development. This remodelling is regulated by a plethora of loosening and stiffening agents such as pectin methyl-esterases, calcium ions, expansins, and glucanases. The tight spatio-temporal regulation of the activities of these agents is a sine qua non condition for proper morphogenesis at cell and tissue levels. The pectin matrix and the cellulose-xyloglucan network operate in concert and their behaviour is mutually dependent on their chemical, structural and mechanical modifications.

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.

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.

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.

The constant force of gravity plays a primordial role in the ontogeny of all living organisms. Plants, for example, develop their roots and shoots in accordance with the direction of the gravitational vector. Any change in the magnitude and/or the direction of gravity has an important impact on the development of tissues and cells. In order to understand how the gravitational force affects plant cell growth and differentiation, we established two complementary experimental procedures with which the effect of hyper-gravity on single plant cell development can be assessed. The single model cell system we used is the pollen tube or male gametophyte which, because of its rapid growth behavior, is known for its instant response to external stresses. The physiological response of the pollen tube can be assessed in a quantitative manner based on changes in the composition and spatial distribution of its cell wall components and in the precisely defined pattern of its very dynamic cytoplasmic streaming. Here, we provide a detailed description of the steps required for the immuno-localization of various cell wall components using microwave-assisted techniques and we explain how live imaging of the intracellular traffic can be achieved under hyper-gravity conditions.

The constant force of gravity plays a primordial role in the ontogeny of all living organisms. Plants, for example, develop their roots and shoots in accordance with the direction of the gravitational vector. Any change in the magnitude and/or the direction of gravity has an important impact on the development of tissues and cells. In order to understand how the gravitational force affects plant cell growth and differentiation, we established two complementary experimental procedures with which the effect of hyper-gravity on single plant cell development can be assessed. The single model cell system we used is the pollen tube or male gametophyte which, because of its rapid growth behavior, is known for its instant response to external stresses. The physiological response of the pollen tube can be assessed in a quantitative manner based on changes in the composition and spatial distribution of its cell wall components and in the precisely defined pattern of its very dynamic cytoplasmic streaming. Here, we provide a detailed description of the steps required for the immuno-localization of various cell wall components using microwave-assisted techniques and we explain how live imaging of the intracellular traffic can be achieved under hyper-gravity conditions.

Cellular organelles move within the cellular volume and the effect of the resulting drag forces on the liquid causes bulk movement in the cytosol. The movement of both organelles and cytosol leads to an overall motion pattern called cytoplasmic streaming or cyclosis. This streaming enables the active and passive transport of molecules and organelles between cellular compartments. Furthermore, the fusion and budding of vesicles with and from the plasma membrane (exo/endocytosis) allow for transport of material between the inside and the outside of the cell. In the pollen tube, cytoplasmic streaming and exo/endocytosis are very active and fulfill several different functions. In this review, we focus on the logistics of intracellular motion and transport processes as well as their biophysical underpinnings. We discuss various modeling attempts that have been performed to understand both long-distance shuttling and short-distance targeting of organelles. We show how the combination of mechanical and mathematical modeling with cell biological approaches has contributed to our understanding of intracellular transport logistics.

Plants are able to sense the magnitude and direction of gravity. This capacity is thought to reside in selected cell types within the plant body that are equipped with specialized organelles called statoliths. However, most plant cells do not possess statoliths, yet they respond to changes in gravitational acceleration. To understand the effect of gravity on the metabolism and cellular functioning of non-specialized plant cells, we investigated a rapidly growing plant cell devoid of known statoliths and without gravitropic behavior, the pollen tube. The effects of hyper-gravity and omnidirectional exposure to gravity on intracellular trafficking and on cell wall assembly were assessed in Camellia pollen tubes, a model system with highly reproducible growth behavior in vitro. Using an epi-fluorescence microscope mounted on the Large Diameter Centrifuge at the European Space Agency, we were able to demonstrate that vesicular trafficking is reduced under hyper-gravity conditions. Immuno-cytochemistry confirmed that both in hyper and omnidirectional gravity conditions, the characteristic spatial profiles of cellulose and callose distribution in the pollen tube wall were altered, in accordance with a dose-dependent effect on pollen tube diameter. Our findings suggest that in response to gravity induced stress, the pollen tube responds by modifying cell wall assembly to compensate for the altered mechanical load. The effect was reversible within few minutes demonstrating that the pollen tube is able to quickly adapt to changing stress conditions.

Future space missions and implementation of permanent bases on Moon and Mars will greatly depend on the availability of ambient air and sustainable food supply. Therefore, understanding the effects of altered gravity conditions on plant metabolism and growth is vital for space missions and extra-terrestrial human existence. In this mini-review we summarize how plant cells are thought to perceive changes in magnitude and orientation of the gravity vector. The particular advantages of several single celled model systems for gravity research are explored and an overview over recent advancements and potential use of these systems is provided.

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.

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.

Walled cells such as those of plants and fungi grow by expanding their cellular surface driven by the intracellular turgor pressure. The generation of a particular cellular shape necessitates the precise spatial control of mechanical properties in the polymer-network forming the cell wall. To model protuberance formation in walled cells we established a finite element model. We aimed to identify the requirements for spatial distribution of mechanical properties in the cell wall that would allow the generation of tubular shapes that agree with experimental observations on the pollen tube, a rapidly elongating plant cell. 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 de-esterified pectin.View Large Image | View Hi-Res Image | Download PowerPoint Slide