Anatomy and Ultrastructure of Galls Induced by Neuroterus quercusbaccarum (Hymenoptera: Cynipidae) on Oak Leaves (Quercus robur)

The structure and ultrastructure of two developmental stages of the spangle gall induced by Neuroterus quercusbaccarum (Hymenoptera, Cynipidae) were investigated using light microscopy (LM), fluorescence microscopy (FM), and transmission (TEM) and scanning (SEM) electron microscopy. The general design of the gall structure was typical of Cynipidae, but some structural features distinguished the spangle gall. Previously undescribed, characteristic multicellular epidermal protuberances with large openings were observed in autumn on the surface of galls. These may facilitate the gas exchange between the atmosphere and the inside of the gall, thus assisting larval respiration. The larval chamber is surrounded by both a sclerenchymatous capsule and numerous cells containing calcium oxalate crystals that may both serve as protective barriers. In young galls, the nutritive tissue is a wall-less protoplasmic mass, potentially easily accessible to young larvae with delicate mandibles. Cell walls only develop at a later stage. The nutritive tissue was found to be rich in proteins and lipids, but starch grains were not observed. Cellular topology suggests that spangle galls grow by anticlinal division of marginal epidermal cells and periclinal division of subepidermal cells. Cellular proliferation (hyperplasia) also occurs in the leaf tissue near the connection with the gall peduncle, which eventually lignifies.

Spatio-Temporal Distribution of Cell Wall Components in the Placentas, Ovules and Female Gametophytes of Utricularia during Pollination

In most angiosperms, the female gametophyte is hidden in the mother tissues and the pollen tube enters the ovule via a micropylar canal. The mother tissues play an essential role in the pollen tube guidance. However, in Utricularia, the female gametophyte surpasses the entire micropylar canal and extends beyond the limit of the integument. The female gametophyte then invades the placenta and a part of the central cell has direct contact with the ovary chamber. To date, information about the role of the placenta and integument in pollen tube guidance in Utricularia, which have extra-ovular female gametophytes, has been lacking. The aim of this study was to evaluate the role of the placenta, central cell and integument in pollen tube pollen tube guidance in Utricularia nelumbifolia Gardner and Utricularia humboldtii R.H. Schomb. by studying the production of arabinogalactan proteins. It was also determined whether the production of the arabinogalactan proteins is dependent on pollination in Utricularia. In both of the examined species, arabinogalactan proteins (AGPs) were observed in the placenta (epidermis and nutritive tissue), ovule (integument, chalaza), and female gametophyte of both pollinated and unpollinated flowers, which means that the production of AGPs is independent of pollination; however, the production of some AGPs was lower after fertilization. There were some differences in the production of AGPs between the examined species. The occurrence of AGPs in the placental epidermis and nutritive tissue suggests that they function as an obturator. The production of some AGPs in the ovular tissues (nucellus, integument) was independent of the presence of a mature embryo sac.

Structural and Nutritional Peculiarities Related to Lifespan Differences on Four Lopesia Induced Bivalve-Shaped Galls on the Single Super-Host Mimosa gemmulata

Super-host plants are elegant models to evaluate the peculiarities of gall structural and nutritional profiles due to the stimuli of distinct gall inducers in temporal and spatial perspectives. Galls induced by congeneric insects, Lopesia spp. (Diptera, Cecidomyiidae) on the same host plant, Mimosa gemmulata Barneby (Fabaceae) were analyzed to estimate if variations of 1 or 2 months in gall lifespans may result in differences over the accumulation of nutritional resources, and their compartmentalization both in cell walls and protoplasm. Mimosa gemmulata hosts four Lopesia-induced galls: the lenticular bivalve-shaped gall (LG) with a 2-month life cycle, the brown lanceolate bivalve-shaped gall (BLG) and the green lanceolate bivalve-shaped gall (GLG) with 3 month-life cycles, and the globoid bivalve-shaped gall (GG) with a 4 month-life cycle. The comparisons among the four Lopesia galls, using anatomical, histometric, histochemical, and immunocytochemical tools, have demonstrated that the longest lifespan of the GG related to its highest increment in structural and nutritional traits compared with the LG, GLG, and BLG. The differences among the tissue stratification and cell wall thickness of the galls with the 2-month and the 3-month lifespans were subtle. However, the GG had thicker cell walls and higher stratification of the common storage tissue, schlerenchymatic layers and typical nutritive tissue than the other three gall morphospecies. The higher tissue thickness of the GG was followed by the formation of a bidirectional gradient of carbohydrates in the protoplasm, and the detection of xyloglucans in cell walls. Current data supported the presumption that the longest the lifespan, the highest the impact over the structural and nutritional metabolism of the Lopesia galls associated to M. gemmulata.

Three in one: evolution of viviparity, coenocytic placenta and polyembryony in cyclostome bryozoans

Background Placentation has evolved multiple times among both chordates and invertebrates. Although they are structurally less complex, invertebrate placentae are much more diverse in their origin, development and position. Aquatic colonial suspension-feeders from the phylum Bryozoa acquired placental analogues multiple times, representing an outstanding example of their structural diversity and evolution. Among them, the clade Cyclostomata is the only one in which placentation is associated with viviparity and polyembryony—a unique combination not present in any other invertebrate group. Results The histological and ultrastructural study of the sexual polymorphic zooids (gonozooids) in two cyclostome species, Crisia eburnea and Crisiella producta, revealed embryos embedded in a placental analogue (nutritive tissue) with a unique structure—comprising coenocytes and solitary cells—previously unknown in animals. Coenocytes originate via nuclear multiplication and cytoplasmic growth among the cells surrounding the early embryo. This process also affects cells of the membranous sac, which initially serves as a hydrostatic system but later becomes main part of the placenta. The nutritive tissue is both highly dynamic, permanently rearranging its structure, and highly integrated with its coenocytic ‘elements’ being interconnected via cytoplasmic bridges and various cell contacts. This tissue shows evidence of both nutrient synthesis and transport (bidirectional transcytosis), supporting the enclosed multiple progeny. Growing primary embryo produces secondary embryos (via fission) that develop into larvae; both the secondary embyos and larvae show signs of endocytosis. Interzooidal communication pores are occupied by 1‒2 specialized pore-cells probably involved in the transport of nutrients between zooids. Conclusions Cyclostome nutritive tissue is currently the only known example of a coenocytic placental analogue, although syncytial ‘elements’ could potentially be formed in them too. Structurally and functionally (but not developmentally) the nutritive tissue can be compared with the syncytial placental analogues of certain invertebrates and chordates. Evolution of the cyclostome placenta, involving transformation of the hydrostatic apparatus (membranous sac) and change of its function to embryonic nourishment, is an example of exaptation that is rather widespread among matrotrophic bryozoans. We speculate that the acquisition of a highly advanced placenta providing massive nourishment might support the evolution of polyembryony in cyclostomes. In turn, massive and continuous embryonic production led to the evolution of enlarged incubating polymorphic gonozooids hosting multiple progeny.

Evolution and ecology of seed internal morphology in relation to germination characteristics in Amaranthaceae

Background and aims Internal seed morphological traits such as embryo characteristics and nutritive tissue can vary considerably within a plant lineage. These traits play a prominent role in germination processes and the success of seedling establishment, and are therefore under high selective pressure, especially in environments hostile to seedlings such as arid, saline or highly dynamic habitats. We investigated the relationships of seed internal morphology and germination characteristics of 84 species of Amaranthaceae s.l., a family with numerous lineages that have adapted to stressful growing conditions. Methods We used seed cross sections to assess embryo type, embryo to seed surface ratio and radicle to cotyledon length ratio. Furthermore, seed mass, mean time to germination, habitat preferences and further plant traits such as C3 or C4 photosynthesis and life form were compiled for each species. Data were analyzed using phylogenetic comparative methods. Key results We found embryo type (λ = 1), log seed mass (λ = 0.86) and embryo to seed size ratio (λ = 0.78) to be evolutionarily stable with an annular embryo as ancestral in the family. Linked to shifts to the three derived embryos types (spiral, horseshoe-shaped and curved) is an increase in root to cotyledon length ratio and a reduction of nutritive tissue. We observed stabilizing selection towards seeds with relatively larger embryos with longer radicles and less nutritive tissue that are able to germinate faster, especially in lineages with C4 photosynthesis and/or salt tolerance. Conclusions We conclude that the evolutionary shift of nutrient storage from perisperm to embryo provides an ecological advantage in extreme environments, because it enables faster germination and seedling establishment. Furthermore, the evolutionary shift towards higher root to cotyledon length ratio especially in small-seeded Amaranthaceae growing in saline habitats can provide an ecological advantage for fast seedling establishment.

Ovule-gall stimulating a large fake fruit on Miconia chamissois Naudin (Melastomataceae): a structural overview

The developmental processes of galls are better known when induced on vegetative organs, while they have hardly ever been described for reproductive ones. Herein, galls induced by Allorhogas uberlandiensis (Hymenoptera) on ovules of Miconia chamissois (Melastomataceae) were analyzed in terms of morphological, anatomical, histochemical and cytological characteristics. Galls are induced on the ovules before fertilization, acting as a physiological seed. Therefore, the ovary grows and develops into a fruit-like gall. The ovule-galls are on average 20 times larger than the seeds, and fruit-like gall volume is on average 5.4 times larger than that of mature fruits. These are related to cell hypertrophy and tissue hyperplasia in the host organ. There is a typical nutritive tissue formation in the ovule-gall with lipid, protein and pectin storage, as well as a storage tissue close to the epidermis. This nutritive tissue shows a smooth endoplasmic reticulum, multivesicular bodies and mitochondria-rich cells. After gall induction, all the ovules degenerate and ovary and hypanthium hypertrophy to form the fruit-like gall. The fruit-like galls form a physical barrier blocking the anthesis, preventing fertilization and keeping only ovule-galls. Our study shows that the A. uberlandensis galling insect may reduce the reproductive success of Miconia chamissois, thus acting as a biological control agent for this host plant population.

Extinct Taxa of Exotestal Seeds Close to Austrobaileyales and Nymphaeales From the Early Cretaceous of Portugal

Early Cretaceous mesofossil floras from Portugal and North America include a surprising diversity of small, bitegmic angiosperm seeds with a hard exotestal seed coat. This study describes six different kinds of these seeds from three Portuguese mesofossil localities; Vale de Agua, Torres Vedras, and especially from Famalicão, which has yielded a flora exceptionally rich in exotestal seeds. All the seeds are almost smooth with a characteristic jigsaw puzzle-shaped surface pattern that is formed from the strongly undulate anticlinal walls of the sclerenchyma cells that comprise the exotesta. Several specimens have internal details preserved, including remains of a cellular nutritive tissue interpreted as endosperm, and a tiny embryo with two rudimentary cotyledons. Based on differences in details of the seed coat, and configuration of hilum and micropyle, the fossil seeds are assigned to six new genera, as six new species: Gastonispermum portugallicum gen. et sp. nov., Pazlia hilaris gen. et sp. nov., Pazliopsis reyi gen. et sp. nov., Reyispermum parvum gen. et sp. nov., Lusitanispermum choffatii gen. et sp. nov. and Silutanispermum kvacekiorum gen. et sp. nov. The characteristic exotestal cells with undulate anticlinal walls, details of the hilar and micropylar region, together with the tiny dicotyledonous embryos with rudimentary cotyledons, suggest close relationships to seeds of Nitaspermum and Tanispermum described previously from Early Cretaceous mesofossil floras from eastern North America. These exotestal seeds from Portugal and North America indicate the presence of diverse extinct early angiosperms close to the lineages that today include extant Austrobaileyales and Nymphaeales.

Rightcania and Kvacekispermum: Early Cretaceous Seeds From Eastern North America and Portugal Provide Further Evidence of the Early Chloranthoid Diversification

Abundant flowers, fruits, seeds and stamens that are closely related to extant Chloranthaceae have been reported from the Early Cretaceous floras of Portugal. Among these are small berries with endotestal seeds assigned to the extinct genera Canrightia and Canrightiopsis. Here we describe two new genera, each including a single species, based on fossil fruits and seeds from the Early Cretaceous of eastern North America and Portugal. Both genera have pendent, orthotropous, bitegmic and endotestal ovules/seeds, in which the endotesta consists of a layer of cubic to palisade-shaped crystal cells with endoreticulate fibrous infillings, a combination of features that also characterize Canrightia and Canrightiopsis and that among extant angiosperms are known only for members of the Chloranthaceae. Rightcania kvacekii gen. et sp. nov. from the early to middle Albian Puddledock mesofossil flora of Virginia, USA, is the first representative in the Early Cretaceous floras of North America of a chloranthaceous fossil related to Canrightia and Canrightiopsis. It has three- to five-seeded fruits very similar to fruits and seeds of Canrightia, also with a pronounced tegmen that probably functioned as a nutritive tissue for the developing embryo. Fruits and seeds of Rightcania are larger than those of Canrightia, and also differ in details of the seed coat. Kvacekispermum rugosum gen. et sp. nov. is rare in the late Aptian to early Albian Portuguese mesofossil flora from Vale de Água. It differs from Canrightiopsis in the coarsely rugulate outer surface of the endotesta and its larger size, but is closely similar in the general structure of seed coat and nutritive tissue. Together, Rightcania and Kvacekispermum provide further evidence of the early diversity achieved by chloranthoid angiosperms before the end of the Early Cretaceous.