The role of intracellular symbiotic bacteria in the amino acid nutrition of embryos from the black bean aphid,Aphis fabae

Aphids are well‐known for their symbiotic relationship with intracellular bacteria of the genus Buchnera (γ‐Proteobacteria). The symbiosis has a nutritional basis in that the bacteria supplement the aphid diet of phloem sap through the provision of essential amino acids. To date, few studies have considered the spatial complexity of the association, particularly the delineation of the symbiosis into embryo and maternal compartments. Here, we generate aposymbiotic (bacteria‐free) embryos of the black bean aphid, Aphis fabae (Scopoli) (Hemiptera: Aphididae), as our experimental model and demonstrate that embryos reared in culture media require an external supply of essential amino acids. Analysis of individual amino acid deletions from the culture medium indicate that the key individual amino acids for embryo growth are phenylalanine and valine derived from the maternal tissues, and tryptophan derived from Buchnera. These results are discussed in relation to our current limited understanding of nutrient supply to aphid embryos.     

Viviparity and the reproductive ecology of clinid fishes (Clinidae) from temperate Australian waters

Synopsis Viviparity is reported in twelve species of the family Clinidae in Tasmanian waters, and superfoetation (the simultaneous development of multiple batches of eggs and/or embryos) in nine of the twelve (all in the subfamily Clinini). The reproductive anatomy and life history features are documented in two superfoetate, shallow-water species,Heteroclinus perspicillatus andHeteroclinus sp. nov. (Scott's Weedfish). Gestation of embryos is intrafollicular and their mode of nutrition shifts from lecithotrophy during early development to trophodermal matrotrophy and, eventually, intestinal nutrient transfer. The sizes of the newly born larvae differ between species, but in all cases maternal investment in individual offspring (as measured by the proportionate increase in embryo dry weight)—20125%, 5744% and 3140% forH. perspicillatus, Scott's Weedfish andH. heptaeolus, respectively — is among the highest recorded for teleosts. Relative fecundity, however, is comparable to those of temperate, demersal spawning blennioids, and this suggests that high levels of maternal investment and relatively advanced stages of larval development at parturition for the sub-family as a whole do not involve a cost in individual fecundity. Parturition, which occurs over several months in spring and summer, is asynchronous. After parturition, the larva spend from 3 to 7 weeks in the plankton, primarily in in-shore waters. Post-settlement growth is rapid and most of the reproductive populations consist of fishes in the 1 + year-class.     

Plant potassium nutrition in ectomycorrhizal symbiosis: properties and roles of the three fungal TOK potassium channels in Hebeloma cylindrosporum

Ectomycorrhizal fungi play an essential role in the ecology of boreal and temperate forests through the improvement of tree mineral nutrition. Potassium (K⁺) is an essential nutrient for plants and is needed in high amounts. We recently demonstrated that the ectomycorrhizal fungus Hebeloma cylindrosporum improves the K⁺ nutrition of Pinus pinaster under shortage conditions. Part of the transport systems involved in K⁺ uptake by the fungus has been deciphered, while the molecular players responsible for the transfer of this cation towards the plant remain totally unknown. Analysis of the genome of H. cylindrosporum revealed the presence of three putative tandem‐pore outward‐rectifying K⁺ (TOK) channels that could contribute to this transfer. Here, we report the functional characterization of these three channels through two‐electrode voltage‐clamp experiments in oocytes and yeast complementation assays. The expression pattern and physiological role of these channels were analysed in symbiotic interaction with P. pinaster. Pine seedlings colonized by fungal transformants overexpressing two of them displayed a larger accumulation of K⁺ in shoots. This study revealed that TOK channels have distinctive properties and functions in axenic and symbiotic conditions and suggested that HcTOK2.2 is implicated in the symbiotic transfer of K⁺ from the fungus towards the plant .     

Effects of maternal and embryo characteristics on post-fertilization provisioning in fishes of the genus <Emphasis Type="Italic">Gambusia</Emphasis>

Maternal provisioning of embryos in Gambusia (Poeciliidae) entails both production of large, yolky eggs and mother-to-embryo transfer of nutrients, the latter of which is readily quantified using injection of radiolabeled nutrients. We assayed patterns of nutrient transfer in broods of 26 Gambusia geiseri and 23 Gambusia affinis females, using injection of tritiated leucine. We examined maternal and embryo characteristics affecting the instantaneous rate of transfer and characterized the pattern of transfer to individual embryos within broods. Maternal (female size and condition) and brood characteristics (mean embryo size, developmental stage, brood size) did not predict the mean level of nutrient transfer to embryos in a brood for either species. Within broods, individual provisioning of embryos was not related to developmental stage, but was related to embryo mass in G. affinis with nutrient transfer higher to larger embryos. In addition, overall within-brood variation in nutrient transfer, measured as coefficient of variation in embryo radioactivity, was higher in G. affinis than in G. geiseri.     

Reproductive diapause and the bacterial symbiosis in the sycamore aphid Drepanosiphum platanoidis

1. Parthenogenetic adults of the sycamore aphid Drepanosiphum platanoidis exhibited reproductive diapause for 4–6 weeks in the summer. 2. The diapausing aphids had small gonads (accounting for just 13% of the total aphid protein content) bearing small (< 0.5 mm long) and developmentally immature embryos. There was no evidence of embryo resorption. 3. The diapausing aphids had significantly depressed essential amino acid content and elevated glutamine content, relative to reproductively active D. platanoidis. 4. The reproductive characteristics and amino acid titres of the diapausing aphids resemble those of aphids lacking functional symbiotic bacteria. Uncoupling of maternal and embryo growth and suppression of bacterial function are proposed as key elements in the diapause of D. platanoidis.     

Maternal impact on egg development in Lymnaea stagnalis: a growth factor is produced by the albumen gland in the reproductive tract

Summary

In the freshwater snail Lymnaea stagnalis, perivitellin fluid is the main source of nutrition for developing embryos; it contains predominantly galactogen and proteins. The fluid is produced by the albumen gland, a large exocrine organ in the female reproductive tract. To further define the protein products of this gland and provide more information about the maternal contribution to egg development, albumen glands were extracted and the extracts purified by reversed-phase HPLC. One major HPLC peak fraction exhibited neurotrophic activity when bioassayed on identified Lymnaea neurons in vitro; it has been partially sequenced and has the highest degree of sequence identity with epidermal growth factor (EGF). These results demonstrate that a growth factor with neurotrophic activity is produced by the albumen gland, and is packaged along with the eggs to serve a growth-promoting function in the embryo.     

Two tomato non-symbiotic haemoglobin genes are differentially expressed in response to diverse changes in mineral nutrient status

The role of plant non‐symbiotic haemoglobins remains undefined, but recent findings suggest a possible role in plant nitrate nutrition. This study sought to characterize patterns of gene expression for two tomato (Lycopersicon esculentum L.) non‐symbiotic haemoglobin genes, and to examine how diverse changes in nutrient status influences the expression of these two genes. The deduced amino acid sequences of the genes examined, SOLly GLB1 and SOLly GLB2, are 55% identical. SOLly GLB1 mRNA is present in highest abundance in roots and older stems, whereas SOLly GLB2 mRNA abundance is highest in leaves, but detectable in other tissues including roots. The SOLly GLB1 mRNA levels increased rapidly in roots in response to each nutrient treatment examined in hydroponic culture, including the individual removal of phosphate, potassium and iron from the culture medium, as well as by the addition of nitrate. In contrast, the levels of SOLly GLB2 mRNA were not significantly altered in response to these same treatments. These results are the first report that non‐symbiotic haemoglobin gene expression can be influenced by a broad range of changes both in mineral macronutrient and micronutrient status, suggesting a previously unrecognized generic role for non‐symbiotic haemoglobins in processes associated with mineral nutrient nutrition.     

A novel pattern of placental leucine transfer during mid to late gestation in a highly placentotrophic viviparous lizard

Placentotrophy is the nourishment of embryos by resources provided via the placenta during gestation. The magnitude and timing of placental nutrient support during pregnancy are important for embryonic growth, especially in highly placentotrophic animals such as mammals. However, no study has yet investigated how placental organic nutrient support may change during pregnancy in highly placentotrophic viviparous reptiles. Amino acids are essential nutrients for embryonic growth and leucine is a common amino acid. The magnitude and timing of placental leucine transfer may affect embryonic growth and mass and, therefore, offspring phenotype. In this study, female Pseudemoia entrecasteauxii, a highly placentotrophic viviparous skink, were collected throughout gestation. We injected (3)H-leucine into these gravid females and assessed the transfer of (3)H-leucine into maternal compartments (i.e., the blood and the liver), and into embryonic compartments (i.e., the embryo, the yolk, and the amniotic fluid). At either 60 or 120 min post-injection, the radioactivity in each sample was extracted and then counted, and the transfer ratio was calculated. Our results provide direct evidence that circulating maternal leucine passes through the placenta into the embryos in this species. The relative rate of placental leucine transfer did not alter during mid to late gestation. This suggests the steady somatic growth of the embryos during mid-late pregnancy is dependent upon the placental transfer of nutrients rather than yolk stores. This pattern of placental nutrient support may determine offspring body size at birth and, therefore, offspring fitness in P. entrecasteauxii.     

The effect of undernutrition on the establishment of pregnancy in the ewe

The relationship between nutrition and reproduction in sheep has been the subject of research in several international groups. This review will particularly focus on the effects of undernutrition on the potential causes of reproductive failure including abnormalities of the ovum or the embryo, luteal inadequacy and failure of the supply of progesterone to the uterus, or the mechanisms involved in maternal recognition of pregnancy. The level of nutrition and peripheral progesterone concentrations are inversely related, and increased rates of embryo loss, associated with higher progesterone concentrations in ewes with low levels of nutrition have been reported. Undernutrition may act through changes in the distribution of progesterone in the endometrium. Thus, lower endometrial levels on day 5 of the cycle in ewes fed half of their maintenance requirements have been observed, providing a link between the known role of progesterone in embryo survival by the modulation of uterine function and the higher embryo losses found in undernourished ewes. The evidence of an effect of maternal nutrition on IFNtau secretion from the conceptus and of PGF2alpha production from the uterus is presented. Moreover, undernutrition provokes a reduction in the sensitivity of the endometrium to progesterone that may affect embryo survival. Finally, a state of undernutrition induces changes in the endometrial sensitivity to steroid hormones at early stages of pregnancy that could adversely alter uterine environment to the detriment of embryo survival.     

Injection of essential amino acids substitutes for bacterial supply in aposymbiotic pea aphids (Acyrthosiphon pisum)

The symbiotic bacteria Buchnera contribute to the nutrition of pea aphids, Acyrthosiphon pisum, through the provision of essential amino acids which are lacking in the diet. However, chemically defined diets, containing nutritionally adequate amounts of essential amino acids, fail to rescue aposymbiotic aphids, in which the bacteria have been disrupted with antibiotics. In this study the injection of a mixture of essential amino acids into the haemocoel of aposymbiotic aphids was shown to alleviate, at least partially, the impact of symbiont loss. Specifically, the total amino acid content in the tissues of aposymbiotic aphids was reduced by approximately 40% to levels comparable with symbiotic insects, and there was a 1.7-fold increase in the number of embryos, suggesting that the availability of essential amino acids promotes aphid protein synthesis by rejuvenating the free amino acid pool of aposymbiotic aphids. In addition, a similar effect on the total amino acid content was observed when phenylalanine alone, but not glutamine, lysine or tryptophan, was injected into the haemocoel of aposymbiotic aphids, and there was also a significant increase in the number of embryos following injection of phenylalanine or tryptophan alone. The impact of amino acid injection on the embryo complement of aposymbiotic aphids was limited to an increase in the number of embryos, with no increase in basal embryo size. It is proposed that older embryos may rely on their own complement of symbiotic bacteria for essential amino acid provisioning. Taken together, the data highlight the importance of bacterial provisioning of essential amino acids, particularly the aromatic amino acids, in the intact symbiosis.     

Pathogen infection and selection on fertilization success in Cucumis sativus

We investigated whether resistant pollen genotypes can be selected for when the maternal plants are infected and whether infection can result to changes in the pistil nutrient level influencing reproductive outcome. Both resistance and susceptibility come with costs that may affect pre- and post-fertilization reproductive functions. We performed the study by crossing zucchini yellow mosaic virus resistant and susceptible pollen (from a hybrid donor) to infected and healthy maternal plants. We also analysed the pistil nutrient content in both treatments and found an increase of the susceptible and not resistant genotypes when maternal plants were infected. The level of nutrients K, P and S was higher in the pistils of the infected maternal plants than the healthy ones. Pistil nutrient level did not affect pollen tube growth rates, as indicated by seed siring patterns along the fruit. We propose that the effect on the siring ability of pollen carrying the susceptible and resistant alleles occurred at the post-fertilization stage, possibly as an indirect result of different growth rates of the two embryo genotypes under elevated nutrient conditions. We discuss our results with respect to possibilities of differential selection, costs and reproductive implications.     

Advances in understanding the physiological mechanism of maternal immune tolerance to the embryo

The results of immunological studies, especially research conducted in the last decade, have shown that free (not bound to protein) progesterone molecules (fP₄) can block the ability of dendritic cells, monocytes and macrophages to present antigens to Th cells and thereby reduce maternal immune activity and increase the maternal tolerance to the semiallogenic embryo. Endocrine studies have shown that fP₄ in the female reproductive organs are transferred at high concentrations into the arterial blood that supplies the oviduct and uterus due to the special adaptations of the blood circulation and lymph flow. Here, the authors present the results of numerous studies documenting their thesis that an important element of maternal tolerance of the semiallogenic embryo in the uterus is conditioned by the close interaction of two processes that occur in the reproductive organs: (1) the local decrease of maternal immune system activity, in which the ability of dendritic cells, macrophages and monocytes to present embryonic antigens to Th cells is blocked by fP₄; and (2) the proper function of the system governing the local retrograde and destination transfer of hormones, which increases the concentration of fP₄ that are able to immediately bind to their receptors in dendritic cells and in the monocytes and macrophages present in the blood supplying the oviduct and uterus. The authors believe that the local interaction of the immune and endocrine systems in the female reproductive tract reduces local maternal immunoreactivity and thus fulfils a critical physiological role; this mechanism protects the embryo but does not change the general immunological resistance of the mother.     

The interaction between maternal stress and the ontogeny of the innate immune system during teleost embryogenesis: implications for aquaculture practice

The barrier defences and acellular innate immune proteins play critical roles during the early‐stage fish embryos prior to the development of functional organ systems. The innate immune proteins in the yolk of embryos are of maternal origin. Maternal stress affects the maternal‐to‐embryo transfer of these proteins and, therefore, environmental stressors may change the course of embryo development, including embryonic immunocompetency, via their deleterious effect on maternal physiology. This review focuses on the associations that exist between maternal stress, maternal endocrine disturbance and the responses of the acellular innate immune proteins of early‐stage fish embryos. Early‐stage teleostean embryos are dependent upon the adult female for the formation of the zona pellucida as an essential barrier defence, for their supply of nutrients, and for the innate immunity proteins and antibodies that are transferred from the maternal circulation to the oocytes; maternally derived hormones are also transferred, some of which (such as cortisol) are known to exert a suppressive action on some aspects of the immune defences. This review summarizes what is known about the effects of oocyte cortisol content on the immune system components in early embryos. The review also examines recent evidence that embryonic cells during early cleavage have the capacity to respond to increased maternal cortisol transfer; this emphasizes the importance of maternal and early immune competence on the later life of fishes, both in the wild and in intensive culture.     

The influence of nuptial feeding and sperm transfer on the immunological cost of reproduction in the ground cricket Allonemobius socius

Mating is often accompanied by decreased female immune function across numerous animals systems, suggesting that immune suppression is a widespread reproductive cost. However, the trade‐off between immunity and reproduction can be minimized when females have access to abundant nutrient resources. This observation suggests that the nuptial gifts provided by males in many insect systems may help to offset the common immunological cost to reproduction. In the present study, this hypothesis is tested in the ground cricket Allonemobius socius (Scudder), whose females receive a sizeable haemolymph‐based gift. Accordingly, male gift donation is controlled by covering the tibial spur (the source of the gift) of randomly chosen males with clear nail polish. The influence of sperm transfer on female immunity is disentangled from that of the nuptial gift by also examining females who fail to receive sperm during mating (spermatophore transfer has a 40% failure rate in virgin males). It is predicted that females who receive a nuptial gift will exhibit superior immune function compared with those who receive no gift. The results show that sperm transfer reduces female immune function, which is an expected immunological cost of reproduction. By contrast to the prediction, nuptial gifts do not minimize the immunological cost of reproduction in this system. Unexpectedly, the receipt of a gift appears to decrease female immune function independent of sperm transfer. The findings suggest that the nuptial gift, similar to sperm, signals the female to begin her reproductive investment, causing limited resources to be reallocated from immune function.     

Gametogenesis and embryonic brooding in the cheilostome bryozoan Celleporella hyalina

Celleporella hyalina (L.) is unusual among cheilostomes in producing sexually dimorphic gonozooids which are frontally budded from a layer of sterile autozooids, and also in the possession of a placental system for extracoelomic nutrition of the developing embryo. This paper investigates these two aspects of the reproductive strategy, combining observations of living colonies with ultrastructural study of preserved material. The formation of the male and female gonozooids is described. Female zooids underwent a maximum of four successive reproductive cycles before senescence. About 29% of embryos failed to develop to completion and were prematurely expelled from the ovicell. Spermatogenesis follows the typical cheilostome pattern. Fertilization is internal, and probably occurs during vitellogenesis of the oocyte. Oogenesis involves the activity of a nurse cell. The telolecithal egg increases in volume approximately 15–fold after transfer to the ovicell, and is nourished by a relatively simple placental system formed by the distal epithelium of the maternal zooid.     

An examination of surface epithelium structures of the embryo across the genus Poeciliopsis (Poeciliidae)

In viviparous, teleost fish, with postfertilization maternal nutrient provisioning, embryonic structures that facilitate maternal‐fetal nutrient transfer are predicted to be present. For the family Poeciliidae, only a handful of morphological studies have explored these embryonic specializations. Here, we present a comparative morphological study in the viviparous poeciliid genus, Poeciliopsis . Using microscopy techniques, we examine the embryonic surface epidermis of Poeciliopsis species that vary in their level of postfertilization maternal nutrient provisioning and placentation across two phylogenetic clades and three independent evolutionary origins of placentation. We focus on surface features of the embryo that may facilitate maternal‐fetal nutrient transfer. Specifically, we studied cell apical‐surface morphology associated with the superficial epithelium that covers the body and sac (yolk and pericardial) of embryos at different developmental stages. Scanning electron microscopy revealed common surface epithelial cells across species, including pavement cells with apical‐surface microridges or microvilli and presumed ionocytes and/or mucus‐secreting cells. For three species, in the mid‐stage embryos, the surface of the body and sac were covered in microvillus epithelium. The remaining species did not display microvillus epithelium at any of the stages examined. Instead, their epithelium of the body and sac were composed of cells with apical‐surface microridges. For all species, in the late stage embryos, the surface of the body proper was composed of apical‐surface microridges in a “fingerprint‐like arrangement.” Despite the differences in the surface epithelium of embryos across Poeciliopsis species and embryonic developmental stages, this variation was not associated with the level of postfertilization maternal nutrient provisioning. We discuss these results in light of previous morphological studies of matrotrophic, teleost fish, phylogenetic relationships of Poeciliopsis species, and our earlier comparative microscopy work on the maternal tissue of the Poeciliopsis placenta.     

An examination of the variation in maternal placentae across the genus Poeciliopsis (Poeciliidae)

Placentae show considerable diversity in a number of nonmammalian, viviparous organisms, including amphibians, reptilian sauropsids, teleost fish, and chondrichthyes. However, the evolutionary processes driving the evolution of placenta are still debated. In teleost fishes, the genus Poeciliopsis (Poeciliidae) offers a rare opportunity for studying placental evolution: extensive placentation has evolved three independent times within the last 750,000 years and there is substantial interspecific variation in the degree of embryonic, maternal nutrient provisioning and development of the placenta. In poeciliids, the placenta is composed of a hypertrophied maternal follicular epithelium apposed to a highly vascularized embryonic pericardial sac. To better understand placental evolution, we have undertaken a comprehensive comparative study of the maternal follicle in eight closely related Poeciliopsis species that span the range in postfertilization, embryonic, maternal nutrient provisioning (from lecithotrophs, to moderate matrotrophs, to extensive matrotrophs). Using light and scanning electron microscopy, we found that the species that provide extensive postfertilization maternal nutrient provisioning (extensive matrotrophs) have thicker follicles and more extensive folding of the follicular epithelium compared to the lecithotrophs and moderate matrotrophs. Follicle sections and histology revealed that epithelial folds of the extensive matrotrophs are comprised primarily of cuboidal and columnar cells and are richly supplied with capillaries. Among the extensive matrotrophs, enhancements of follicle traits corresponded with increases in the level of maternal nutrient provisioning. Hypertrophied maternal follicles with richly vascularized folds can serve to increase the surface area and, thus, facilitate the transfer of substances between the mother and developing embryo. Finally, we found egg envelopes in the lecithotrophs and moderate matrotrophs, but not in the extensive matrotrophs. Morphological studies, like this one, can provide a better understanding of the natural variation in the structure and functioning of maternal and offspring traits associated with matrotrophy and, thus, insights into the processes driving placental evolution. J. Morphol. 276:707–720, 2015. © 2015 Wiley Periodicals, Inc.     

Buchnera aphidicola of the birch blister aphid,Hamamelistes betulinus (Horváth, 1896) (Insecta,Hemiptera, Aphididae: Hormaphidinae): molecular characterization,transmission between generations and its geographic significance

The birch blister aphid Hamamelistes betulinus, like most aphids, is host to obligate symbiotic bacterium Buchnera aphidicola. Ultrastructural and molecular analyses did not reveal the presence of secondary symbionts in the body of H. betulinus. The bacteria Buchnera aphidicola are transmitted to the next generation vertically (maternally). The bacteria released from the cytoplasm of the bacteriocyte to the haemolymph migrate to the embryo at the cellular blastoderm stage, through the opening at its posterior pole. Next, the bacteria enter the cytoplasm of newly formed bacteriocytes. The concept of the relationship between the geographic distribution of Hormaphidini aphids and the presence/absence of bacterium Buchnera aphidicola is discussed.     

Maternal source of variability in the embryo development of an annual killifish

Organisms inhabiting unpredictable environments often evolve diversified reproductive bet‐hedging strategies, expressed as production of multiple offspring phenotypes, thereby avoiding complete reproductive failure. To cope with unpredictable rainfall, African annual killifish from temporary savannah pools lay drought‐resistant eggs that vary widely in the duration of embryo development. We examined the sources of variability in the duration of individual embryo development, egg production and fertilization rate in Nothobranchius furzeri. Using a quantitative genetics approach (North Carolina type II design), we found support for maternal effects rather than polyandrous mating as the primary source of the variability in the duration of embryo development. The number of previously laid eggs appeared to serve as an internal physiological cue initiating a shift from rapid‐to‐slow embryo developmental mode. In annual killifish, extensive phenotypic variability in progeny traits is adaptive, as the conditions experienced by parents have limited relevance to the offspring generation. In contrast to genetic control, with high phenotypic expression and heritability, maternal control of traits under natural selection prevents standing genetic diversity from potentially detrimental effects of selection in fluctuating environments.     

Nutrient transfer from soil nematodes to plants: a direct pathway provided by the mycorrhizal mycelial network

A pathway for the transfer of nutrients from dead nematodes to mycorrhizal plants is described for the first time. Plants of Betula pendula were grown in transparent microcosms in the mycorrhizal (M) or non‐mycorrhizal (NM) condition, either with or without nematode necromass of known nitrogen (N) and phosphorus (P) contents as the major potential source of these elements. Plants colonized by the mycorrhizal fungus Paxillus involutus produced greater yields and had larger N and P contents in the presence of nematodes than did their NM counterparts. The symbiotic systems were shown to exploit the N and P originally contained in necromass more effectively, and to transfer the nutrients to the plants in quantities approximately double those seen in NM systems. Even so, NM plants obtained sufficient N and P from dead nematodes to enable some enhancement of growth. Our observations confirm that mycorrhizal fungi provide the potential for the recycling of nutrients contained in this quantitatively important component of the soil mesofauna and demonstrate that the symbiotic pathway is considerably more effective than that provided by saprotrophs alone. The consequences of this nutrient transfer pathway for nutrient recycling in temperate forest ecosystems are considered.