Hydraulic tradeoffs underlie enhanced performance of polyploid trees under soil water deficit.

The relationships between aerial organ morpho-anatomy of woody polyploid plants with their functional hydraulics under water stress remain largely understudied. We evaluated growth-associated traits, aerial organ xylem anatomy, and physiological parameters of diploid, triploid, and tetraploid genotypes of atemoyas (Annona cherimola × Annona squamosa), which belong to the woody perennial genus Annona (Annonaceae), testing their performance under long-term soil water reduction. The contrasting phenotypes of vigorous triploids and dwarf tetraploids consistently showed stomatal size-density tradeoff. The vessel elements in aerial organs were ∼1.5 times wider in polyploids compared with diploids, and triploids displayed the lowest vessel density. Plant hydraulic conductance was higher in well-irrigated diploids while their tolerance to drought was lower. The phenotypic disparity of atemoya polyploids associated with contrasting leaf and stem xylem porosity traits that coordinate to regulate water balances between the trees and the belowground and aboveground environments. Polyploid trees displayed better performance under soil water scarcity, and consequently, could present more sustainable agricultural and forestry genotypes to cope with water stress.

Changes in ploidy affect vascular allometry and hydraulic function in Mangifera indica trees.

The enucleated vascular elements of the xylem and the phloem offer an excellent system to test the effect of ploidy on plant function because variation in vascular geometry has a direct influence on transport efficiency. However, evaluations of conduit sizes in polyploid plants have remained elusive, most remarkably in woody species. We used a combination of molecular, physiological and microscopy techniques to model the hydraulic resistance between source and sinks in tetraploid and diploid mango trees. Tetraploids exhibited larger chloroplasts, mesophyll cells and stomatal guard cells, resulting in higher leaf elastic modulus and lower dehydration rates, despite the high water potentials of both ploidies in the field. Both the xylem and the phloem displayed a scaling of conduits with ploidy, revealing attenuated hydraulic resistance in tetraploids. Conspicuous wall hygroscopic moieties in the cells involved in transpiration and transport indicate a role in volumetric adjustments as a result of turgor change in both ploidies. In autotetraploids, the enlargement of organelles, cells and tissues, which are critical for water and photoassimilate transport at long distances, point to major physiological novelties associated with whole-genome duplication.

Sieve tube structural variation in Austrobaileya scandens and its significance for lianescence.

Lianas combine large leaf areas with slender stems, features that require an efficient vascular system. The only extant member of the Austrobaileyaceae is an endemic twining liana of the tropical Australian forests with well-known xylem hydraulics, but the vascular phloem continuum aboveground remains understudied. Microscopy analysis across leaf vein orders and stems of Austrobaileya scandens revealed a low foliar xylem:phloem ratio, with isodiametric vascular elements along the midrib, but tapered across vein orders. Sieve plate pore radii increased from 0.08 µm in minor veins to 0.12 µm in the petiole, but only to 0.20 µm at the stem base, tens of metres away. In easily bent searcher branches, phloem conduits have pectin-rich walls and simple plates, whereas in twining stems, conduits were connected through highly angled and densely porated sieve plates. The hydraulic resistance of phloem conduits in the twisted and elongated stems of A. scandens is large compared with trees of similar stature; phloem hydraulic resistance decreases from leaves to stems, consistent with the efficient delivery of photoassimilates from sources under Münch predictions. Sink strength of a continuously growing canopy might be stronger than in self-supporting understory plants, favoring resource allocation to aerial organs and the attainment of vertical stature.

Idioblasts and peltate hairs as distribution networks for water absorbed by xerophilous leaves.

Capparis odoratissima is a tree species native to semi-arid environments of South America where low soil water availability coexists with frequent night-time fog. A previous study showed that water applied to leaf surfaces enhanced leaf hydration, photosynthesis and growth, but the mechanisms of foliar water uptake are unknown. Here, we combine detailed anatomical evaluations with water and dye uptake experiments in the laboratory, and use immunolocalization of pectin and arabinogalactan protein epitopes to characterize water uptake pathways in leaves. Abaxially, the leaves of C. odoratissima are covered with peltate hairs, while the adaxial surfaces are glabrous. Both surfaces are able to absorb condensed water, but the abaxial surface has higher rates of water uptake. Thousands of idioblasts per cm2 , a higher density than stomata, connect the adaxial leaf surface and the abaxial peltate hairs, both of which contain hygroscopic substances such as arabinogalactan proteins and pectins. The highly specialized anatomy of the leaves of C odoratissima fulfils the dual function of minimizing water loss when stomata are closed, while maintaining the ability to absorb liquid water. Cell-wall related hygroscopic compounds in the peltate hairs and idioblasts create a network of microchannels that maintain leaf hydration and promote water uptake.

Not all 'pine cones' flex: functional trade-offs and the evolution of seed release mechanisms.

Seed dispersal is critical for plants, but the evolution of mechanisms that actually release seeds from their parents is not well understood. We use the reproductive cones of conifers, specifically the Pinaceae clade, to explore the factors driving the evolution of different release mechanisms in plants. We combine comparative anatomical and phylogenetic analyses to test whether fundamental trade-offs in the mechanical and hydraulic properties of vasculature underlie the evolution of two seed release mechanisms: cone scale flexion and cone scale shedding. We then test whether these mechanisms are linked with differences in seed size, dispersal syndrome and reproductive allocation. Cone scale xylem in flexing species is tough, but poorly conductive. Xylem in shedding species is less extensive, fragile and highly conductive; its thin-walled tracheids allow scales to easily fracture at maturity. Shedding is also consistently associated with large, densely packed seeds. Pinaceae cones exploit a well-known trade-off in xylem mechanical strength vs hydraulic efficiency to generate release mechanisms that allow seeds of various sizes to leave the protecting cone. The linkage among release mechanisms, vascular anatomy and seed traits illustrates how a wide variety of selective pressures may influence the function and physiology of reproductive structures.

Scaling of phloem hydraulic resistance in stems and leaves of the understory angiosperm shrub Illicium parviflorum.

PREMISE OF THE STUDY: Recent studies in canopy-dominant trees revealed axial scaling of phloem structure. However, whether this pattern is found in woody plants of the understory, the environment of most angiosperms from the ANA grade (Amborellales-Nymphaeales-Austrobaileyales), is unknown. METHODS: We used seedlings and adult plants of the understory tropical shrub Illicium parviflorum, a member of the lineage Austrobaileyales, to explore the anatomy and physiology of the phloem in their aerial parts, including changes through ontogeny. KEY RESULTS: Adult plants maintain a similar proportion of phloem tissue across stem diameters, but larger conduit dimensions and number cause the hydraulic resistance of the phloem to decrease toward the base of the plant. Small sieve plate pores resulted in an overall higher sieve tube hydraulic resistance than has been reported in other woody angiosperms. Sieve elements increase in size from minor to major leaf veins, but were shorter and narrower in petioles. The low carbon assimilation rates of seedlings and mature plants contrasted with a 3-fold higher phloem sap velocity in seedlings, suggesting that phloem transport velocity is modulated through ontogeny. CONCLUSIONS: The overall architecture of the phloem tissue in this understory angiosperm shrub scales in a manner consistent with taller trees that make up the forest canopy. Thus, the evolution of larger sieve plate pores in canopy-dominant trees may have played a key role in allowing woody angiosperms to extend beyond their understory origins.

Why are the seed cones of conifers so diverse at pollination?

Background and Aims: Form and function relationships in plant reproductive structures have long fascinated biologists. Although the intricate associations between specific pollinators and reproductive morphology have been widely explored among animal-pollinated plants, the evolutionary processes underlying the diverse morphologies of wind-pollinated plants remain less well understood. Here we study how this diversity may have arisen by focusing on two conifer species in the pine family that have divergent reproductive cone morphologies at pollination. Methods: Standard histology methods, artificial wind pollination assays and phylogenetic analyses were used in this study. Key Results: A detailed study of cone ontogeny in these species reveals that variation in the rate at which their cone scales mature means that pollination occurs at different stages in their development, and thus in association with different specific morphologies. Pollination experiments nevertheless indicate that both species effectively capture pollen. Conclusions: In wind-pollinated plants, morphological diversity may result from simple variation in development among lineages rather than selective pressures for any major differences in function or performance. This work also illustrates the broader importance of developmental context in understanding plant form and function relationships; because plant reproductive structures perform many different functions over their lifetime, subtle differences in development may dramatically alter the specific morphologies that they use to meet these demands.

Prolonged embryogenesis in Austrobaileya scandens (Austrobaileyaceae): its ecological and evolutionary significance.

The embryology of basal angiosperm lineages (Amborella, Nymphaeales and Austrobaileyales) is central to reconstructing the early evolution of flowering plants. Previous studies have shown that mature seeds in Austrobaileyales are albuminous, with a small embryo surrounded by a substantial diploid endosperm. However, little is known of seed ontogeny and seedling germination in Austrobaileya scandens, sister to all other extant Austrobaileyales. Standard histochemical techniques were used to study ovule/seed development and germination of Austrobaileya. Early development of the endosperm in Austrobaileya is ab initio cellular with pronounced cell proliferation. The nucellus transiently accumulates some starch, but is obliterated by expansion of a massive endosperm, where all embryo-nourishing reserves are ultimately stored. Twelve months elapse from fertilization to fruit abscission. Seeds are dispersed with a minute embryo, requiring 12 additional months for seedling establishment. The 2 yr required for seedling establishment is an apomorphic feature of Austrobaileya, probably related to germination in extremely dark understory conditions. Remarkably, although Austrobaileya seeds are nearly 50 times larger (by length) than the smallest seeds of extinct and extant members of early divergent angiosperm lineages, the embryo to seed ratio (E : S) falls squarely within the narrow range that characterizes the albuminous seeds of ancient flowering plant lineages.

Pollen tube access to the ovule is mediated by glycoprotein secretion on the obturator of apple (Malus × domestica, Borkh).

Background and Aims: Within the ovary, the obturator bridges the pathway of the pollen tube from the style to the ovule. Despite its widespread presence among flowering plants, its function has only been studied in a handful of species, and the molecules involved in pollen tube-obturator cross-talk have not been explored hitherto. This work evaluates the involvement of glucans and glycoproteins on pollen tube growth in the obturator of apple flowers ( Malus × domestica) . Methods: Pollen tube kinetics were sequentially examined in the pistil and related to changes occurring on the obturator using histochemistry and inmunocytochemistry. To discriminate between changes in the obturator induced by pollen tubes from those developmentally regulated, both pollinated and unpollinated pistils were examined. Key Results: Pollen tube growth rates were slow in the stigma, faster in the style and slow again in the ovary. The arrival of pollen tubes at the obturator was concomitant with the secretion of proteins, saccharides and glycoprotein epitopes belonging to extensins and arabinogalactan proteins (AGPs). While some of these secretions - extensins and AGPs labelled by JIM13 - were developmentally regulated, others - AGPs labelled by JIM8 - were elicited by the presence of pollen tubes. Following pollen tube passage, all these glycoproteins were depleted. Conclusions: The results show a timely secretion of glycoproteins on the obturator surface concomitant with pollen tube arrival at this structure. The fact that their secretion is depleted following pollen tube passage strongly suggests their role in regulating pollen tube access to the ovule. Remarkably, both the regulation of the secretion of the different glycoproteins, as well as their association with the performance of pollen tubes exhibit similarities with those observed in the stigma, in line with their common developmental origin.

Pollen-pistil interaction in pawpaw (Asimina triloba), the northernmost species of the mainly tropical family Annonaceae.

PREMISE OF THE STUDY: The pawpaw, Asimina triloba, is an underutilized fruit crop native to North America that belongs to the mainly tropical, early-divergent family Annonaceae. Asimina is the only genus within the Annonaceae with species adapted to cold climates. A thorough analysis of its reproductive biology, specifically pollen-pistil interaction during the progamic phase, is essential to understand both its adaptation to cold climates and how to optimize its fertilization and fruit set. METHODS: We characterized pollen-pistil interaction in Asimina triloba, including the floral cycle and anatomy, stigmatic receptivity, and the pollen tube pathway. We used a combination of histological, cytological, and immunolocalization approaches. KEY RESULTS: Asimina triloba has a gynoecium formed by plicate carpels with a short stylar canal. Unicellular papillae form a continuous tissue covered by a copious secretion from the stigma to the ovary, which is most prominent on the stigma surface where it forms an extragynoecial compitum. Compared to the stigmas of other species in the Annonaceae, the stigmas of A. triloba show a long stigmatic receptivity associated with a long flowering cycle. Stigmatic receptivity is concomitant with the secretion of cell-wall-related arabinogalactan proteins (AGPs). CONCLUSIONS: A long female phase with a long period of stigmatic receptivity is unusual among protogynous flowers of the magnoliid clade, suggesting a derived condition of A. triloba within the Annonaceae. This phase further correlates with the presence of cell-wall-related arabinogalactan proteins in the secretion, which may indicate the conservation of these glycoproteins during stigmatic receptivity and pollen tube growth in angiosperms.

Floral biology and ovule and seed ontogeny of Nymphaea thermarum, a water lily at the brink of extinction with potential as a model system for basal angiosperms.

BACKGROUND AND AIMS: Nymphaea thermarum is a member of the Nymphaeales, of one of the most ancient lineages of flowering plants. This species was only recently described and then declared extinct in the wild, so little is known about its reproductive biology. In general, the complete ontogeny of ovules and seeds is not well documented among species of Nymphaea and has never been studied in the subgenus Brachyceras, the clade to which N. thermarum belongs. METHODS: Flowers and fruits were processed for brightfield, epifluorescence and confocal microscopy. Flower morphology, with emphasis on the timing of male and female functions, was correlated with key developmental stages of the ovule and the female gametophyte. Development of the seed tissues and dynamics of polysaccharide reserves in the endosperm, perisperm and embryo were examined. KEY RESULTS: Pollen release in N. thermarum starts before the flower opens. Cell walls of the micropylar nucellus show layering of callose and cellulose in a manner reminiscent of transfer cell wall patterning. Endosperm development is ab initio cellular, with micropylar and chalazal domains that embark on distinct developmental trajectories. The surrounding maternal perisperm occupies the majority of seed volume and accumulates starch centrifugally. In mature seeds, a minute but fully developed embryo is surrounded by a single, persistent layer of endosperm. CONCLUSIONS: Early male and female function indicate that N. thermarum is predisposed towards self-pollination, a phenomenon that is likely to have evolved multiple times within Nymphaea. While formation of distinct micropylar and chalazal developmental domains in the endosperm, along with a copious perisperm, characterize the seeds of most members of the Nymphaeales, seed ontogenies vary between and among the constituent families. Floral biology, life history traits and small genome size make N. thermarum uniquely promising as an early-diverging angiosperm model system for genetic and molecular studies.

Glycoprotein composition along the pistil of Malus x domestica and the modulation of pollen tube growth.

BACKGROUND: The characteristics of pollen tube growth are not constant, but display distinct patterns of growth within the different tissues of the pistil. In the stigma, the growth rate is slow and autotrophic, whereas in the style, it is rapid and heterotrophic. Very little is known about the interactions between these distinct maternal tissues and the traversing pollen tube and the role of this interaction on the observed metabolism. In this work we characterise pollen tube growth in the apple flower and look for differences in glycoprotein epitope localization between two different maternal tissues, the stigma and the style. RESULTS: While immunocytochemically-detected arabinogalactan proteins were present at high levels in the stigma, they were not detected in the transmitting tissue of the style, where extensins were abundant. Whereas extensins remained at high levels in unpollinated pistils, they were no longer present in the style following pollen tube passage. Similarily, while abundant in unpollinated styles, insoluble polysaccharides such as ß-glucans, were depleted in pollinated pistils. CONCLUSIONS: The switch from autotropic to heterotrophic pollen tube growth correlates spatially with a change of glycoprotein epitopes between the stigma and the style. The depletion of extensins and polysaccharides following pollen tube passage in the style suggest a possible contribution to the acceleration of heterotrophic pollen tube growth, which would imply an active contribution of female tissues on prezygotic male-female crosstalk.

Arabinogalactan proteins mark stigmatic receptivity in the protogynous flowers of Magnolia virginiana (Magnoliaceae).

PREMISE OF THE STUDY: Factors affecting floral receptivity in angiosperms remain opaque, but recent studies suggest that the acquisition of stigmatic receptivity associated with cell-wall-related arabinogalactan proteins (AGPs) may be a widespread feature of flowering plants. Here, the time during which a stigma is receptive is evaluated and related to the secretion of AGPs in Magnolia virginiana, a protogynous member of an early-divergent angiosperm clade (magnoliids) with a clearly discernible female receptive phase. METHODS: Magnolia virginiana flower phenology was documented, and histochemical changes in the stigma before and after pollination were examined. Stigmatic receptivity was evaluated in relation to the secretion of AGPs detected in whole mounts and immunolocalized in sectioned stigmas. KEY RESULTS: Protogynous Magnolia flowers had a precise window of stigmatic receptivity, which is concomitant with the secretion of two AGPs labeled for different epitopes. After pollen germination and tube growth, these two AGPs could no longer be detected in the stigmas, suggesting that these AGPs interact with the growing male gametophytes and could be markers of stigmatic receptivity. CONCLUSIONS: These results show that the period of stigmatic receptivity is finely coordinated with the secretion of two arabinogalactan proteins on stigmas of flowers of M. virginiana. This first report of AGP presence in stigmatic tissues in a member of the magnoliids, together with recently described similar patterns in eudicots, monocots, and members of early-divergent lineages of flowering plants, suggests an ancient and widespread role for AGPs on stigmatic receptivity in angiosperms.

Concluding Embryogenesis After Diaspora: Seed Germination in Illicium Parviflorum.

Albuminous seeds, dispersed with a minimally developed embryo surrounded by nutrient storage tissue, are pervasive across extinct and extant early diverging angiosperm lineages. Typically, seed ontogenic studies have focused on the time between fertilization and seed release, but in albuminous seeds, embryogenesis is incomplete at the time of seed dispersal. Here, I studied the morphological and nutritional relationships between the embryo and the endosperm after seed dispersal in Illicium parviflorum (Austrobaileyales). Seeds of I. parviflorum germinate over a period of three months. Different stages during the germination process were anatomically evaluated using a combination of histochemistry and immunocytochemistry. At dispersal, the seeds of Illicium contain a tiny achlorophyllous embryo with minimal histological differentiation, surrounded by copious amounts of lipo-protein globules stored in the endosperm within cell walls rich in un-esterified pectins. Six weeks later, the embryo expanded and differentiated the vascular tissues before the emergence of the radicle through the seed coat, as the stored lipids and proteins coalesced within cells. Six weeks later, the cotyledons contained starch and complex lipids intracellularly, and accumulated low-esterified pectins in their cell walls. The proteolipid-rich albuminous seeds of Illicium exemplify how woody angiosperms of the Austrobaileyales, Amborellales, and many magnoliids release seeds with high-energy storage compounds that are reprocessed by embryos that complete development during germination. Seedlings of these lineages thrive in the understory of tropical environments, which match with the predicted habitats where angiosperms evolved.

How dense can you be? New automatic measures of vein density in angiosperm leaves.

Premise: Because of the trade-off between water loss and carbon dioxide assimilation, the conductivity of the transpiration path in a leaf is an important limit on photosynthesis. Closely packed veins correspond to short paths and high assimilation rates while widely spaced veins are associated with higher resistance to flow and lower maximum photosynthetic rates. Vein length per area (VLA) has become the standard metric for comparing leaves with different vein densities; its measurement typically utilizes digital image processing with varying amounts of human input. Methods and Results: Here, we propose three new ways of measuring vein density using image analysis that improve on currently available procedures: (1) areole area distributions, (2) a sizing transform, and (3) a distance map. Each alternative has distinct practical, statistical, and biological limitations and advantages. In particular, we advocate the log-transformed modal distance map of a vein mask as an estimator to replace VLA as a standard metric for vein density. Conclusions: These methods, for which open-source code appropriate for high-throughput automation is provided, improve on VLA by producing determinate measures of vein density as distributions rather than point estimates. Combined with advances in image quality and computational efficiency, these methods should help clarify the physiological and evolutionary significance of vein density.

Foliar Pectins and Physiology of Diploid and Autotetraploid Mango Genotypes under Water Stress.

The cultivation of mango in Mediterranean-type climates is challenged by the depletion of freshwater. Polyploids are alternative genotypes with potential greater water use efficiency, but field evaluations of the anatomy and physiology of conspecific adult polyploid trees under water stress remain poorly explored. We combined field anatomical evaluations with measurements of leaf water potential (Ψl) and stomatal conductance (Gs) comparing one diploid and one autotetraploid tree per treatment with and without irrigation during dry summers (when fruits develop). Autotetraploid leaves displayed lower Ψl and Gs in both treatments, but the lack of irrigation only affected Gs. Foliar cells of the adaxial epidermis and the spongy mesophyll contained linear pectin epitopes, whereas branched pectins were localized in the abaxial epidermis, the chloroplast membrane, and the sieve tube elements of the phloem. Cell and fruit organ size was larger in autotetraploid than in diploid mango trees, but the sugar content in the fruits was similar between both cytotypes. Specific cell wall hygroscopic pectins correlate with more stable Ψl of autotetraploid leaves under soil water shortage, keeping lower Gs compared with diploids. These preliminary results point to diploids as more susceptible to water deficits than tetraploids.

Arabinogalactan-protein secretion is associated with the acquisition of stigmatic receptivity in the apple flower.

BACKGROUND AND AIMS: Stigmatic receptivity plays a clear role in pollination dynamics; however, little is known about the factors that confer to a stigma the competence to be receptive for the germination of pollen grains. In this work, a developmental approach is used to evaluate the acquisition of stigmatic receptivity and its relationship with a possible change in arabinogalactan-proteins (AGPs). METHODS: Flowers of the domestic apple, Malus × domestica, were assessed for their capacity to support pollen germination at different developmental stages. Stigmas from these same stages were characterized morphologically and different AGP epitopes detected by immunocytochemistry. KEY RESULTS: Acquisition of stigmatic receptivity and the secretion of classical AGPs from stigmatic cells occurred concurrently and following the same spatial distribution. While in unpollinated stigmas AGPs appeared unaltered, in cross-pollinated stigmas AGPs epitopes vanished as pollen tubes passed by. CONCLUSIONS: The concurrent secretion of AGPs with the acquisition of stigmatic receptivity, together with the differential response in unpollinated and cross-pollinated pistils point out a role of AGPs in supporting pollen tube germination and strongly suggest that secretion of AGPs is associated with the acquisition of stigma receptivity.

Conductivity of the phloem in mango (Mangifera indica L.).

Mango (Mangifera indica L., Anacardiaceae), the fifth most consumed fruit worldwide, is one of the most important fruit crops in tropical regions, but its vascular anatomy is quite unexplored. Previous studies examined the xylem structure in the stems of mango, but the anatomy of the phloem has remained elusive, leaving the long-distance transport of photoassimilates understudied. We combined fluorescence and electron microscopy to evaluate the structure of the phloem tissue in the tapering branches of mango trees, and used this information to describe the hydraulic conductivity of its sieve tube elements following current models of fluid transport in trees. We revealed that the anatomy of the phloem changes from current year branches, where it was protected by pericyclic fibres, to older ones, where the lack of fibres was concomitant with laticiferous canals embedded in the phloem tissue. Callose was present in the sieve plates, but also in the walls of the phloem sieve cells, making them discernible from other phloem cells. A scaling geometry of the sieve tube elements-including the number of sieve areas and the pore size across tapering branches-resulted in an exponential conductivity towards the base of the tree. These evaluations in mango fit with previous measurements of the phloem architecture in the stems of forest trees, suggesting that, despite agronomic management, the phloem sieve cells scale with the tapering branches. The pipe model theory applied to the continuous tubing system of the phloem appears as a good approach to understand the hydraulic transport of photoassimilates in fruit trees.

Arabinogalactan proteins mediate intercellular crosstalk in the ovule of apple flowers.

KEY MESSAGE: AGP-rich glycoproteins mediate pollen-ovule interactions and cell patterning in the embryo sac of apple before and after fertilization. Glycoproteins are significant players in the dialog that takes place between growing pollen tubes and the stigma and style in the angiosperms. Yet, information is scarce on their possible involvement in the ovule, a sporophytic organ that hosts the female gametophyte. Apple flowers have a prolonged lapse of time between pollination and fertilization, offering a great system to study the developmental basis of glycoprotein secretion and their putative role during the last stages of the progamic phase and early seed initiation. For this purpose, the sequential pollen tube elongation within the ovary was examined in relation to changes in arabinogalactan proteins (AGPs) in the tissues of the ovule before and after fertilization. To evaluate what of these changes are developmentally regulated, unpollinated and pollinated flowers were compared. AGPs paved the pollen tube pathway in the ovules along the micropylar canal, and the nucellus entrance toward the synergids, which also developmentally accumulated AGPs at the filiform apparatus. Glycoproteins vanished from all these tissues following pollen tube passage, strongly suggesting a role in pollen-ovule interaction. In addition, AGPs marked the primary cell walls of the haploid cells of the female gametophyte, and they further built up in the cell walls of the embryo sac and developing embryo, layering the interactive walls of the three generations hosted in the ovule, the maternal sporophytic tissues, the female gametophyte, and the developing embryo.

Phloem networks in leaves.

The survival of all vascular plants depends on phloem and xylem, which comprise a hydraulically coupled tissue system that transports photosynthates, water, and a variety of other molecules and ions. Although xylem hydraulics has been extensively studied, until recently, comparatively little is known quantitatively about the phloem hydraulic network and how it is functionally coupled to the xylem network, particularly in photosynthetic leaves. Here, we summarize recent advances in quantifying phloem hydraulics in fully expanded mature leaves with different vascular architectures and show that (1) the size of phloem conducting cells across phylogenetically different taxa scales isometrically with respect to xylem conducting cell size, (2) cell transport areas and lengths increase along phloem transport pathways in a manner that can be used to model Münch's pressure-flow hypothesis, and (3) report observations that invalidate da Vinci's and Murray's hydraulic models as plausible constructs for understanding photosynthate transport in the leaf lamina.