Nutritive Tissue Research Articles

Integrated insights into the cytological, histochemical, and cell wall composition features of Espinosa nothofagi (Hymenoptera) gall tissues: implications for functionality.

Many insect-induced galls are considered complex structures due to their tissue compartmentalization and multiple roles performed by them. The current study investigates the complex interaction between Nothofagus obliqua host plant and the hymenopteran gall-inducer Espinosa nothofagi, focusing on cell wall properties and cytological features. The E. nothofagi galls present an inner cortex with nutritive and storage tissues, as well as outer cortex with epidermis, chlorenchyma, and water-storing parenchyma. The water-storing parenchyma cells are rich in pectins, heteromannans, and xyloglucans in their walls, and have large vacuoles. Homogalacturonans contribute to water retention, and periplasmic spaces function as additional water reservoirs. Nutritive storage cell walls support nutrient storage, with plasmodesmata facilitating nutrient mobilization crucial for larval nutrition. Their primary and sometimes thick secondary cell walls support structural integrity and act as a carbon reserve. The absent labeling of non-cellulosic epitopes indicates a predominantly cellulosic nature in nutritive cell walls, facilitating larval access to lipid, protein, and reducing sugar-rich contents. The nutritive tissue, with functional chloroplasts and high metabolism-related organelles, displays signs of self-sufficiency, emphasizing its role in larval nutrition and cellular maintenance. Overall, the intricate cell wall composition in E. nothofagi galls showcases adaptations for water storage, nutrient mobilization, and larval nutrition, contributing significantly to our understanding of plant-insect interactions.

Short-Term Periodic Fasting Reduces Ischemia-Induced Necrosis in Musculocutaneous Flap Tissue.

Periodic fasting (PF) as a form of dietary restriction has been shown to induce tissue-protective effects against ischemic injury in several different tissues. Accordingly, in this study we analyzed whether a short-term 24 h fast is suitable to prevent necrosis of musculocutaneous flap tissue undergoing acute persistent ischemia. C57BL/6N mice were randomly divided into a PF group (n = 8) and a control group that was given unrestricted access to standard chow (n = 8). The PF animals underwent a 24 h fast immediately before flap elevation and had unrestricted access to food for the rest of the 10 day observation period. Musculocutaneous flaps with a random pattern design were dissected on the animals' backs and mounted into dorsal skinfold chambers. On days 1, 3, 5, 7 and 10 after surgery, nutritive tissue perfusion, angiogenesis and flap necrosis were evaluated using intravital fluorescence microscopy. Thereafter, the flap tissue was excised and fixed for histological and immunohistochemical analyses. The flaps of PF-treated animals exhibited a higher functional capillary density and more newly formed microvessels, resulting in a significantly increased flap survival rate. Moreover, they contained a lower number of myeloperoxidase (MPO)-positive neutrophilic granulocytes and cleaved caspase-3-positive apoptotic cells in the transition zone between vital and necrotic flap tissue. These findings indicate that short-term PF improves tissue survival in ischemically challenged musculocutaneous flaps by maintaining nutritive blood perfusion and dampening ischemia-induced inflammation.

Cell wall epitope distribution in the functional compartments of galls induced by Palaeomystella oligophaga (Lepidoptera) in Macairea radula (Melastomataceae)

Context The parasitic interaction between the galling insect Palaeomystella oligophaga (Lepidoptera) and the host plant tissues of Macairea radula (Melastomataceae) leads to the formation of globoid galls. These galls have storage and typical nutritive tissues (outer and inner compartments, respectively), whose functions may be related to differential cell wall component distributions and to the stage of development of the gall inducer. Aims We evaluated whether the cell wall composition of the gall compartments (storage and nutritive tissues) changes according to the developmental stage of the gall inducer P. oligophaga. Methods The galls occupied by the gall inducer in different stages of development (i.e. larval and pupal stages, as well as empty galls) were collected and submitted to histological examination, immunocytochemical analyses using monoclonal antibodies for pectins, hemicelluloses, and glycoproteins, and histochemical analyses using phloroglucinol for lignins. Key results The histological results showed that nutritive tissues have smaller cells compared with storage ones when the galling insect is in the larval stage. Immunocytochemical analyses also showed the occurrence of both methyl-esterified and unesterified pectins, as well hemicelluloses in the nutritive tissue during the larval stage. The dynamic of some cell wall components changes when the gall inducer is in the pupal stage, or the galls are empty. Conclusions The changes in the composition of the cell walls, especially regarding homogalacturonans and xyloglucans, support the high metabolism and resource availability in cell walls of nutritive tissue for the gall inducer. Implications The cell wall composition may indicate important metabolic steps during gall formation and galling insect diet.

Nutritive tissue rich in reserves in the cell wall and protoplast: the case of Manihot esculenta (Euphorbiaceae) galls induced by Iatrophobia brasiliensis (Diptera, Cecidomyiidae).

The galls can offer shelter, protection, and an adequate diet for the gall-inducing organisms. Herein, we evaluated the structure of Manihot esculenta leaves and galls induced by Iatrophobia brasiliensis in order to identify metabolic and cell wall composition changes. We expected to find a complex gall with high primary metabolism in a typical nutritive tissue. Non-galled leaves and galls were subjected to anatomical, histochemical, and immunocytochemical analyses to evaluate the structural features, primary and secondary metabolites, and glycoproteins, pectins, and hemicelluloses in the cell wall. The gall is cylindric, with a uniseriate epidermis, a larval chamber, and a parenchymatic cortex divided into outer and inner compartments. The outer compartment has large cells with intercellular spaces and stocks starch and is designated as storage tissue. Reducing sugars, proteins, phenolic compounds, and alkaloids were detected in the protoplast of inner tissue cells of galls, named nutritive tissue, which presents five layers of compact small cells. Cell walls with esterified homogalacturonans (HGs) occurred in some cells of the galls indicating the continuous biosynthesis of HGs. For both non-galled leaves and galls, galactans and xyloglucans were broadly labeled on the cell walls, indicating a cell growth capacity and cell wall stiffness, respectively. The cell wall of the nutritive tissue had wide labeling for glycoproteins, HGs, heteroxylans, and xyloglucans, which can be used as source for the diet of the galling insect. Manihot esculenta galls have compartments specialized in the protection and feeding of the galling insect, structured by nutritive tissue rich in resource compounds, in the cell walls and protoplast.

Vascular implications of Dasineura sp. galls' establishment on Peumus boldus stems.

Some chewing larvae are capable of inducing galls in the host vascular cylinder, e.g. Dasineura sp. (Cecidomyiidae) on Peumus boldus stems. Due to the medicinal and economic importance of P. boldus, the anatomical and functional implications of establishment of Dasineura sp. on P. boldus stems were investigated. We asked if establishment of Dasineura sp. in P. boldus stems induces abnormalities at the cellular and organizational level of the vascular system that increase during gall development in favour of the hydric status of the gall. Anatomical alterations induced in the stems during gall development were determined. Cytohistometric analyses in mature galls were compared to non-galled stems, and water potential and leaf area of non-galled stems were compared with galled stems. Dasineura sp. establishes in the vascular cambium, leading to delignification and rupture of xylem cells, inhibiting formation of phloem and perivascular sclerenchyma. Gall diameter increases together with larval feeding activity, producing a large larval chamber and numerous layers of nutritive tissue, vascular parenchyma, and sclerenchyma. These anatomical alterations do not affect the leaf area of galled stems but favour increased water flow towards these stems. The anatomical alterations induced by Dasineura sp. in P. boldus stems guarantee water and nutrient supply to the gall and larva. After the inducer exits stems, some host branches no longer have vascular connections with the plant body.

Microvascular Fragments Protect Ischemic Musculocutaneous Flap Tissue from Necrosis by Improving Nutritive Tissue Perfusion and Suppressing Apoptosis.

Microvascular fragments (MVF) derived from enzymatically digested adipose tissue are functional vessel segments that have been shown to increase the survival rate of surgical flaps. However, the underlying mechanisms have not been clarified so far. To achieve this, we raised random-pattern musculocutaneous flaps on the back of wild-type mice and mounted them into dorsal skinfold chambers. The flaps were injected with MVF that were freshly isolated from green fluorescent protein-positive (GFP+) donor mice or saline solution (control). On days 1, 3, 5, 7, and 10 after surgery, intravital fluorescence microscopy was performed for the quantitative assessment of angiogenesis, nutritive blood perfusion, and flap necrosis. Subsequently, the flaps were analyzed by histology and immunohistochemistry. The injection of MVF reduced necrosis of the ischemic flap tissue by ~20%. When compared to controls, MVF-injected flaps also displayed a significantly higher functional capillary density and number of newly formed microvessels in the transition zone, where vital tissue bordered on necrotic tissue. Immunohistochemical analyses revealed a markedly lower number of cleaved caspase-3+ apoptotic cells in the transition zone of MVF-injected flaps and a significantly increased number of CD31+ microvessels in both the flaps' base and transition zone. Up to ~10% of these microvessels were GFP+, proving their origin from injected MVF. These findings demonstrate that MVF reduce flap necrosis by increasing angiogenesis, improving nutritive tissue perfusion, and suppressing apoptosis. Hence, the injection of MVF may represent a promising strategy to reduce ischemia-induced flap necrosis in future clinical practice.

"The phenomenon of autonomous endosperm in sexual and apomictic plants".

Endosperm is a key nutritive tissue that supports the developing embryo or seedling, and serves as a major nutritional source for human and livestock feed. In sexual flowering plants, it generally develops after fertilization. However, autonomous endosperm (AE) formation (i.e., independently from fertilization) is also possible. Recent findings of AE loci/ genes and aberrant imprinting in native apomicts, together with a successful initiation of parthenogenesis in rice and lettuce, have enhanced our understanding of the mechanisms bridging sexual and apomictic seed formation. However, the mechanisms driving AE development are not well understood. This review presents novel aspects related to AE development in sexual and asexual plants underlying stress conditions as the primary trigger for AE. Both, application of hormones to unfertilized ovules and mutations that impair epigenetic regulation lead to AE development in sexual Arabidopsis thaliana, which may point to a common pathway for both phenomena. Apomictic-like AE development under experimental conditions can take place due to auxin-dependent gene expression and/or DNA methylation.

A broad spectrum of host plant responses to the actions of the gall midge: case study of Robinia pseudoacacia L. and Obolodiplosis robiniae (Haldeman)

This study aims to provide insights into plant-insect interaction during the formation and development of open gall structure on the leaves of Robinia pseudoacacia during gall formation by Obolodiplosis robiniae. This was the first time such far-reaching studies were performed at a biochemical and anatomical level. The gall wall is created from a few thick cells covered with epidermis. This parenchymatous nutritive tissue is rich in starch. Sclerenchyma only occurs around the vascular bundles as a result of the lignification of the parenchyma of the bundle sheaths. The level of reactive oxygen species (ROS) in the new structure was reduced and catalase activity was inhibited, which suggests another pathway of ROS decomposition – e.g. by ascorbate or glutathione peroxidase. The gall structure was combined with an increasing level of protein and non-protein thiols. Phenols seems to be a good protective factor; whose level was lower in infected leaflets. Levels of MUFA (monosaturated fatty acids) and SFA (saturated fatty acids) rose, probably as source of food for insects. The amount of fatty acid is positively correlated with the plant response. We detected that non infected leaflets produced C6:0 (hexanoic acid) and C8:0 (octanoic acid) fatty acids connected with odor. Changes in gall color as they develop are connected with photosynthetic pigments degradation (mainly chlorophylls) where the pathway of astaxanthin transformation to fatty acid is considered to be the most important process during gall maturation. Nutritive tissue is composed mainly of octadecanoic acid (C18:0) – a main source of food for O. robiniae.

Alterations induced by Tortrimosaica polypodivora on the stems of Microgramma vacciniifolia: simple or complex galls?

Abstract Microgramma vacciniifolia, an epiphyte fern, hosts of two stem galls. One is induced by Tortrimosaica polypodivora (Lepidoptera), which can also induce galls in M. squamulosa and M. mortoniana. The alterations induced by T. polypodivora on M. vacciniifolia stem were compared to non-galled organs to evaluate the anatomical potentials of host ferns in response to a galling Lepidoptera. Histochemical and histometrical comparisons between galled and non-galled stems were performed to assess the processes leading to gall formation. M. vacciinifolia and M. squamulosa galls were anatomically similar, although their sizes differ, reflecting the growth potential of each host species. Simple structural alterations, such as hyperplasia of cortical and pericycle cells, occur during gall formation, while cell hypertrophy, common in more complex galls, was only detected on the pericycle. Meristele size remained unaltered in galls, but the pericycle appeared hyperplasic. The protective scales were broader in galls. A nutritive tissue with lipids, typical to Lepidoptera galls, was observed around the larval chamber, with small cells and meristematic activity. Starch, proteins, and reducing sugars accumulated in nutritive cells are uncommonly found in Lepidoptera galls. Despite simple structural alterations, T. polypodivora induced a gradient of primary metabolites, similar to angiosperm galls.

As soft as silk: structural and chemical traits can help with the identification of Niphidium crassifolium (Polypodiaceae) gall inducers

Abstract Ferns have been poorly reported as hosts of gall inducers, and their multitrophic interactions and relationships are practically unknown to science. We focused on Niphidium crassifolium (Polypodiaceae) that hosts globoid leaf galls. The galls on N. crassifolium have only reported for the South and Southeast regions of Brazil, with a discussion regarding the identity of the gall inducer: is it a Cecidomyiidae-Diptera or to Coccidae-Hemiptera? These two insect groups have distinct characteristics and consequently their galls must have distinct anatomical and histochemical traits. Such traits may work out as functional tools to be used to confirm the taxa of the associated galling herbivore and to evaluate their geographic distribution. Our study aimed to expand the known distribution of N. crassifolium and its interactions based on an inventory of scientific articles and on herbarium data, and also to test the usefulness of anatomical and histochemical traits for proposing the identity of the gall inducer. The geographic distribution of N. crassifolium galls involves five South American countries, i.e., Bolivia, French Guiana, Guyana, Peru, and Brazil. The development of an outer tissue compartment with phenolic-rich cell layers and an inner nutritive tissue leads us to infer that the inducer belongs to the Cecidomyiidae family.

Gall-inducing Eriophyes tiliae stimulates the metabolism of Tilia platyphyllos leaves towards oxidative protection

Red galls have high levels of anthocyanins which perform different physiological functions, such as antioxidants and protection against UVB radiation. High levels of anthocyanins and other polyphenols have been associated with low photosynthetic pigment content. In environments with high levels of UVB radiation, it would thus be expected that red galls would have high anthocyanin and polyphenol levels and low photosynthetic pigment contents, enabling the gall with high antioxidant capacity compared to its host organ. The red galls induced by Eriophyes tiliae, and their host environment of Tilia platyphyllos leaves in the Mediterranean climate of Chile, were investigated in relation to their anatomy, histochemistry, pigment, sugar, protein, and polyphenol contents, and antioxidant capacity. The anthocyanin, sugars, and polyphenol contents and the antioxidant capacity were increased in galls. Photosynthetic pigment and protein contents were higher in non-galled leaves. The high levels of anthocyanin and total polyphenols increase the galls' antioxidant capacity in the high UV radiation environment of a Mediterranean climate. The establishment of E. tiliae induced redifferentiation of nutritive tissue, rich in sugars, proteins, and lipids, and an inner epidermis with trichomes and long emergences. E. tiliae galls’ structural and metabolic features are probably enhanced towards mite nutrition and protection. The current results shed light on the role of anthocyanin in the antioxidant protection of plant galls in environments with high UV irradiance.

Perioperative Intermittent Fasting Protects Ischemic Musculocutaneous Flap Tissue from Necrosis.

Dietary restriction, such as intermittent fasting (IF), has previously been shown to protect various tissues from ischemia-induced necrosis. Based on this finding, the authors herein evaluated for the first time the tissue-protective effects of IF in musculocutaneous flaps. Male C57BL/6N mice were randomly assigned to an IF group ( n = 8) and a control group with unrestricted access to standard diet ( n = 8). IF animals were put on a perioperative feeding schedule with 8 hours of unrestricted access to standard diet per day starting 7 days before flap elevation up to 3 days after surgery. Random pattern musculocutaneous flaps were raised and mounted into a dorsal skinfold chamber. Intravital fluorescence microscopy was performed on days 1, 3, 5, 7, and 10 after surgery for the quantitative assessment of angiogenesis, nutritive blood perfusion, and flap necrosis. After the in vivo observation period, the flaps were harvested for additional histologic and immunohistochemical analyses. The authors found that the IF group exhibited more newly formed microvessels and a higher functional capillary density within the flap tissue when compared with controls. This was associated with a significantly lower rate of tissue necrosis. Immunohistochemical detection of different inflammatory cell subtypes revealed a markedly reduced number of invading myeloperoxidase-positive neutrophilic granulocytes in the musculocutaneous tissue of IF-treated animals. Considered together, these findings indicate that IF prevents flap necrosis by maintaining nutritive tissue perfusion and suppressing ischemia-induced inflammation. Given the frequent use of flaps in plastic surgery and the high rates of ischemic complications, any new approach to decrease such complications is clinically relevant.

Endosperm cellularization failure induces a dehydration-stress response leading to embryo arrest.

The endosperm is a nutritive tissue supporting embryo growth in flowering plants. Most commonly, the endosperm initially develops as a coenocyte (multinucleate cell) and then cellularizes. This process of cellularization is frequently disrupted in hybrid seeds generated by crosses between different flowering plant species or plants that differ in ploidy, resulting in embryo arrest and seed lethality. The reason for embryo arrest upon cellularization failure remains unclear. In this study, we show that triploid Arabidopsis thaliana embryos surrounded by uncellularized endosperm mount an osmotic stress response that is connected to increased levels of abscisic acid (ABA) and enhanced ABA responses. Impairing ABA biosynthesis and signaling aggravated triploid seed abortion, while increasing endogenous ABA levels as well as the exogenous application of ABA-induced endosperm cellularization and suppressed embryo growth arrest. Taking these results together, we propose that endosperm cellularization is required to establish dehydration tolerance in the developing embryo, ensuring its survival during seed maturation.

A phenological trick and cell wall bricks toward adaptive strategies of Mimosa tenuiflora-Lopesia mimosae interaction in Caatinga environment

Caatinga environment tends to impose constraints on the phenology and development of the host plants and their associated galls. Phenological synchronism in Mimosa tenuiflora-Lopesia mimosae system, and anatomical, cytometric, and immunocytochemical analyzes reveal strategies of the gall survivorship in Caatinga. As phenological strategies, the peak of young galls synchronizes with leaf sprouting in the rainy season, and the reproductive phase is concomitant to gall senescence in the dry season. As structural strategies, the reorganization and the compartmentalization of gall tissues assist gall development in response to environmental constraints, and the predominance of periclinal cell elongation determined the gall bivalve shape. The distribution of pectins, glycoproteins, and hemicelluloses modulate cell walls toward favoring gall resistance to drought stress. The homogalacturonans confer high porosity to the cell walls of the typical nutritive tissue. The galactans and arabinans confer flexibility to the cell walls of the common storage tissue favoring tolerance to drought. The xylogalacturonans and heteromannans in the cell walls of the typical nutritive tissue are associated with nutritive reserves directed to the galling insect. The phenological synchronism, modulation of leaf tissues and cell wall components confer water and nutritional resources to the Lopesia mimosae microenvironment, the gall.

Bromelain Protects Critically Perfused Musculocutaneous Flap Tissue from Necrosis

Bromelain has previously been shown to prevent ischemia-induced necrosis in different types of tissues. In the present study, we, therefore, evaluated for the first time, the tissue-protective effects of bromelain in musculocutaneous flaps in mice. Adult C57BL/6N mice were randomly assigned to a bromelain treatment group and a control group. The animals were treated daily with intraperitoneal injections of 20 mg/kg bromelain or saline (control), starting 1 h before the flap elevation throughout a 10-day observation period. The random-pattern musculocutaneous flaps were raised on the backs of the animals and mounted into a dorsal skinfold chamber. Angiogenesis, nutritive blood perfusion and flap necrosis were quantitatively analyzed by means of repeated intravital fluorescence microscopy over 10 days after surgery. After the last microscopy, the flaps were harvested for additional histological and immunohistochemical analyses. Bromelain reduced necrosis of the critically perfused flap tissue by ~25%. The bromelain-treated flaps also exhibited a significantly higher functional microvessel density and an elevated formation of newly developed microvessels in the transition zone between the vital and necrotic tissues when compared to the controls. Immunohistochemical analyses demonstrated a markedly lower invasion of the myeloperoxidase-positive neutrophilic granulocytes and a significantly reduced number of cleaved caspase 3-positive apoptotic cells in the transition zone of bromelain-treated musculocutaneous flaps. These findings indicate that bromelain prevents flap necrosis by maintaining nutritive tissue perfusion and by suppressing ischemia-induced inflammation and apoptosis. Hence, bromelain may represent a promising compound to prevent ischemia-induced flap necrosis in clinical practice.

Parasitoid impairment on the galling Lopesia sp. activity reflects on the cytological and histochemical profiles of the globoid bivalve-shaped gall on Mimosa gemmulata.

Gall cytological and histochemical features established by the constant feeding activity of the associated gall inducer may be changed due to the attack of parasitoids. We accessed two tri-trophic systems involving the globoid bivalve-shaped gall on Mimosa gemmulata Barneby (Fabaceae) and its galling undescribed species of Lopesia (Diptera: Cecidomyiidae), which may be ectoparasitized by Torymus sp. (Hymenoptera: Torymidae) or endoparasitized by a polyembryonic Platygastridae (Hymenoptera), as models of study. The ectoparasitoid species paralyzes and kills Lopesia sp. larva, which stops the feeding stimuli, while the endoparasitoid larvae feed in Lopesia sp. larva body and keep it alive for a certain time. Our hypothesis is that the time lapse of Lopesia sp. feeding impairment by the two parasitoids will cause distinct cytological and histochemical responses in the ecto- and endoparasitized galls compared to the non-parasitized condition. In both parasitoidism cases, the impairment of the feeding activity of the galling Lopesia sp. directs the common storage and nutritive cells toward a similar process of induced cell death, involving cell collapse and loss of membrane integrity. The cell metabolism is maintained mainly by mitochondria, and by the translocation of lipids from the common storage tissue, via plasmodesmata, through the living sclereids of the mechanical zone toward the nutritive tissue. Accordingly, the parasitoid impairment on the feeding activity of Lopesia sp. larvae causes precocious senescence, but similar cytological alterations, and no impact over the histochemical profiles, regarding lipids, reactive oxygen species, and secondary metabolites, which support gall metabolism along the parasitoid cycles.

Complex meristematic activity induced by Eucecidoses minutanus on Schinus engleri turns shoots into galls.

Gall-inducing organisms change the development of their host plant organs, resulting in ontogenetic patterns not observed in the non-galled plants. Distinct taxa induce galls on Schinus spp., manipulating meristematic patterns in the host plant in distinct ways. Here we report ontogenetic novelties induced in the lateral buds of S. engleri by Eucecidoses minutanus, a Cecidosidae, whose galls have been poorly understood. The anatomy, histochemistry, and histometry of galls in distinct phases of development, non-galled buds, and stems of Schinus engleri were analyzed in parallel with the instars of E. minutanus to detail the morphogenetic changes in the host with each larval stage. Ontogenetic phases of the galls were intricately associated with larval development. First and second-instar larvae induced pericycle and pith cells to dedifferentiate into the gall inner meristem, where hyperplasia and cell hypertrophy characterized the growth and development phase of the gall. The innermost layers were lipid-rich nutritive cells that lined the larval chamber. Additional vascular bundle rows were produced in young galls. Third and fourth instar-larvae were associated with the gall maturation phase: centripetal lignification of the outer parenchyma cell layers, epidermal stratification, and activation of a cambium-like meristem (CLM). The CLM activity resulted in new layers of nutritive cells that differentiated inward as the first layers of nutritive cells were consumed by E. minutanus larvae, and, also, in more parenchyma cell layers that formed outward. All tissues between the innermost layer of nutritive tissue that surround the gall chamber and the outermost layer of the dermal system that externally covers the gall form the gall wall, and increased in thickness until the end of gall maturation. E. minutanus induces a structurally complex globoid stem gall, modifying all host plant tissues and stimulating a novel meristematic pattern in S. engleri. The gall developmental stages are each related to specific gall-inducing instars, as gall development progresses according to the development of E. minutanus.

The ontogenesis of four Lopesia Rübsaamen (Cecidomyiidae) galls on the super-host Mimosa gemmulata Barneby (Fabaceae) reveals peculiar anatomical traits

The super-host Mimosa gemmulata has four bivalve-shaped galls induced on the pinna-rachis by congeneric Lopesia spp., which are characterized by four distinct morphologies: the lenticular (LG), the green lanceolate (GLG), the brown lanceolate (BLG), and the globoid (GG) galls. The ontogenesis of these distinct Lopesia galls is related to the capacity of each inducer to differentially manipulate the development of the host plant tissues along its lifespan, resulting in specific gall phenotypes. The ontogenesis of these galls reveals that the differentiation of two types of trichomes is a conservative anatomical trait of M. gemmulata. Nevertheless, under the influence of the four Lopesia spp., the trichome types are altered in size, number of cells, and distribution in the dermal system of each gall. Another conservative trait regarding the vascular system is the collateral arrangement in non-galled and galled conditions, but with an increase in GG vascular elements. The nutritive cells of three out of the four galls originate exclusively from the epidermis, while parenchyma cell lineages participate in the formation of the nutritive tissue of the GG. The intrinsic anatomical features of the four Lopesia galls relate to the site of induction and to the original cell lineages recruited toward the ontogenesis of similar bivalve-shaped morphotypes. However, specific ontogenetic deviations determine each Lopesia gall morphospecies. In contrast, the peculiar structural-functional traits of the GG develop due to its longest lifespan, which consequently leads to the highest stimuli-induced responses in all host plant cell lineages.

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

Simple SummaryGalls induced by insects are commonly found on oak leaves in temperate climates. Their induction and development are, however, still only partially known. Our aim was to present a detailed description of their anatomy and fine structure. Using scanning electron microscopy (SEM), we observed the wax cover of a gall and two types of openings on its surface: one similar to degenerated stomata and the other one that we called “large openings on epidermal protuberance”. Nutritive tissue underwent marked changes with time: in a young gall, the cellular walls were not visible, and a distinct cellular structure appeared only later. In the mound of the gall, there was a distinct subepidermal layer of cells containing dense cytoplasm. Tannins occurred in vacuoles or in cell walls in granular form or in compact formations. The leaf subtending the gall showed additional cell divisions and strong lignification of cell walls in the tissue surrounding the peduncle of the gall.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.