RESUMO
Ovule morphology, megasporogenesis, and megagametogenesis processes were examined in Hydrocleys nymphoides, Alisma plantago-aquatica, and Sagittaria montevidensis. Each of these species belongs to a different clade within the Alismataceae family. It is worth mentioning that the genus Hydrocleys previously belonged to the Limnocharitaceae family but is now classified within the Alismataceae. Flowers in different developmental stages were processed following classical histological methods for their observation with bright-field microscope. The three species present an anatropous and bitegmic mature ovule. This is tenuinucellate in A. plantago-aquatica and S. montevidensis and pseudo-crassinucellate in H. nymphoides. Although all three species have the same type of megasporogenesis, they differ in the megagametogenesis and in the total number of nuclei and cells that form the mature gametophyte. H. nymphoides has a female gametophyte composed of four cells and four nuclei, while A. plantago-aquatica and S. montevidensis have a female gametophyte of five cells and six nuclei. The results are discussed according to the phylogenetic position of each of the species. Moreover, new types of megagametophyte development are described: Hydrocleys and Sagittaria types. The reduction of the female gametophyte with respect to the Polygonum type is found in families belonging to the ANA grade and in other aquatic families within the order Alismatales. We infer that the reduction in the number of cells and nuclei in the female gametophyte is characteristic of species that inhabit aquatic environments. Future studies in aquatic species belonging to other families would be necessary to confirm this hypothesis.
Assuntos
Sagittaria , Sagittaria/citologia , Óvulo Vegetal/citologia , Alisma/química , Alisma/citologia , Alismataceae/citologiaRESUMO
KEY MESSAGE: In Araucaria angustifolia, the seed scale is part of the ovule, the female gametophyte presents a monosporic origin and arises from a coenocytic tetrad, and the pollen tube presents a single axis. The seed cone of conifers has many informative features, and its ontogenetic data may help interpret relationships among function, development patterns, and homology among seed plants. We reported the seed cone development, from pollination to pre-fertilization, including seed scale, ovule ontogeny, and pollen tube growth in Araucaria angustifolia. The study was performed using light microscopy, scanning electron microscopy, and X-ray microcomputed tomography (µCT). During the pollination period, the ovule arises right after the seed scale has emerged. From that event to the pre-fertilization period takes about 14 months. Megasporogenesis occurs three weeks after ovule formation, producing a coenocytic tetrad. At the same time as the female gametophyte's first nuclear division begins, the pollen tube grows through the seed scale adaxial face. Until maturity, the megagametophyte goes through the free nuclei stage, cellularization stage, and cellular growth stage. Along its development, many pollen tubes develop in the nucellar tissue extending straight toward the female gametophyte. Our observations show that the seed scale came out of the same primordia of the ovule, agreeing with past studies that this structure is part of the ovule itself. The formation of a female gametophyte with a monosporic origin that arises from a coenocytic tetrad was described for the first time in conifers, and the three-dimensional reconstruction of the ovule revealed the presence of pollen tubes with only one axis and no branches, highlighting a new pattern of pollen tube growth in Araucariaceae.
Assuntos
Araucaria , Araucariaceae , Polinização , Tubo Polínico , Cone de Plantas , Brasil , Microtomografia por Raio-X , Sementes , Óvulo Vegetal , BiologiaRESUMO
Gametophytic self-incompatibility (GSI) has been mainly described in species-rich clades within Orchidaceae. We report GSI for a genus within Maxillariinae, one of the most conspicuous and diverse subtribes of neotropical orchids. Here, we describe the reproductive system of Brasiliorchis picta, B. phoenicanthera, and B. porphyrostele. Anatomical studies of fruit development showed that pollen tubes of aborted, self-pollinated flowers degenerate half-way in the stylar channel and never reach the ovules. Spontaneous self-pollination and emasculation set no fruits for none of the sampled species, thus supporting the hypothesis that these three species studied rely on the agency of pollinators and pollinator-mediated cross-pollination to set fruit. Fruit set from cross-pollinations ranged from 33.4 to 77.5%. One self-pollinated fruit of B. porphyrostele developed to completion. All other fruits aborted between 10 and 21 days after pollination. These data support previous evidence of variable strength GSI being exhibited in orchid species. Additional studies of self-incompatibility systems are needed to evaluate their role in species diversification and evolution of reproductive strategies in Maxillariinae and to allow for effective conservation strategies of threatened orchid species.
Assuntos
Orchidaceae , Pólen , Reprodução , Polinização , Frutas , FloresRESUMO
To explain the co-existence and maintenance of females along with hermaphrodite plants, the female advantage hypothesis has been proposed where females should show greater fecundity compared to their conspecific hermaphrodites. On the other hand, greater attraction would be selected in the hermaphrodites to increase their male function, potentially leading to larger showier flowers, with more rewards. Here, I tested the sexual dimorphism trade-off hypothesis with the gynomonoecious-gynodioecious Dianthus plumarius (Caryophyllaceae), in the gardens of Bariloche (Patagonia, Argentina). I measured in female and hermaphrodite plants: flower size, nectar volume and concentration, flower lifespan, ovule production, seed number, seed set and seed weight. Additionally, bagging and pollen supplementation experiments were carried out to evaluate pollen limitation, probability of apomixis, if spontaneous autogamy is possible, and to examine the importance of pollen origin. I found that hermaphrodite flowers are more attractive, with larger-sized flowers and higher nectar volume, whereas female flowers compensate with longer lifespan of stigmatic receptivity and more concentrated nectar. Despite ovule number was lower in female flowers, these showed higher seed set and produced more and heavier seeds than hermaphrodites under open pollination. No evidence of apomixis was found in females, but spontaneous autogamy may occur in hermaphrodites. Hand-pollination experiments showed first that both flower types suffered pollen limitation, but it was higher on hermaphrodite flowers. Finally, despite self-compatibility, pollen origin is important because hand self-pollination decreases seed weight. These findings provide strong evidence in support of the mechanisms and underlying conditions that would allow the co-existence and maintenance of female and hermaphrodite individuals within populations.
RESUMO
PREMISE: The number of open flowers on a plant (i.e., floral display size) can influence plant fitness by increasing pollinator attraction. However, diminishing marginal fitness returns with increasing floral display are expected as pollinators tend to visit more flowers per plant consecutively. An extended flower visitation sequence increases the fraction of ovules disabled by self-pollination (ovule discounting) and reduces the fraction of a plant's own pollen that is exported to sire seeds in other plants (pollen discounting). Hermaphroditic species with a genetic system that prevents self-fertilization (self-incompatibility) would avoid ovule discounting and its fitness cost, whereas species without such a genetically based barrier would not. Contrarily, pollen discounting would be an unavoidable consequence of a large floral display irrespective of selfing barriers. Nevertheless, the increasing fitness costs of ovule and pollen discounting could be offset by respectively increasing ovule and pollen production per flower. METHODS: We compiled data on floral display size and pollen and ovule production per flower for 1241 animal-pollinated, hermaphroditic angiosperm species, including data on the compatibility system for 779 species. We used phylogenetic general linear mixed models to assess the relations of pollen and ovule production to floral display size. RESULTS: Our findings provide evidence of increasing pollen production, but not of ovule production, with increasing display size irrespective of compatibility system and even after accounting for potentially confounding effects like flower size and growth form. CONCLUSIONS: Our comparative study supports the pollen-discount expectation of an adaptive link between per-flower pollen production and floral display across animal-pollinated angiosperms.
Assuntos
Magnoliopsida , Animais , Magnoliopsida/genética , Filogenia , Pólen/genética , Polinização , Plantas , Flores/genéticaRESUMO
MAIN CONCLUSION: An exonuclease V homologue from apomictic Brachiaria brizantha is expressed and localized in nucellar cells at key moments when these cells differentiate to give rise to unreduced gametophytes. Brachiaria is a genus of forage grasses with economical and agricultural importance to Brazil. Brachiaria reproduces by aposporic apomixis, in which unreduced embryo sacs, derived from nucellar cells, other than the megaspore mother cell (MMC), are formed. The unreduced embryo sacs produce an embryo without fertilization resulting in clones of the mother plant. Comparative gene expression analysis in ovaries of sexual and apomictic Brachiaria spp. revealed a sequence from B. brizantha that showed a distinct pattern of expression in ovaries of sexual and apomictic plants. In this work, we describe a gene named BbrizExoV with strong identity to exonuclease V (Exo V) genes from other grasses. Sequence analysis in signal prediction tools showed that BbrizExoV might have dual localization, depending on the translation point. A longer form to the nucleus and a shorter form which would be directed to the chloroplast. This is also the case for monocot sequences analyzed from other species. The long form of BbrizExoV protein localizes to the nucleus of onion epidermal cells. Analysis of ExoV proteins from dicot species, with exception of Arabidopsis thaliana ExoVL protein, showed only one localization. Using a template-based AlphaFold 2 modelling approach the structure of BbrizExoV in complex with metal and ssDNA was predicted based on the holo structure of the human counterpart. Features predicted to define ssDNA binding but a lack of sequence specificity are shared between the human enzyme and BbrizExoV. Expression analyses indicated the precise site and timing of transcript accumulation during ovule development, which coincides with the differentiation of nucelar cells to form the typical aposporic four-celled unreduced gametophyte. A putative function for this protein is proposed based on its homology and expression pattern.
Assuntos
Apomixia , Arabidopsis , Brachiaria , Humanos , Exodesoxirribonuclease V , Gametogênese Vegetal , Células Germinativas Vegetais , PoaceaeRESUMO
KEY MESSAGE: Inheritance of the presence/absence of seeds in Annona squamosa is mediated by a single fully recessive gene and is caused by a deletion of the INNER NO OUTER (INO) locus. For some fruits, seedless varieties are desirable for consumption and processing. In the sugar apple tree (Annona squamosa L.), the seedless trait in the Thai seedless (Ts) and Brazilian seedless (Bs) accessions was associated with defective ovules and an apparent deletion of the INNER NO OUTER (INO) ovule development gene locus. Segregation analysis of F2 and backcross descendants of crosses of Bs to fertile wild-type varieties in this species with a multi-year generation time showed that seedlessness was recessive and controlled by a single locus. Comparison of whole genome sequence of a wild-type plant and a third accession, Hawaiian seedless (Hs), identified a 16 kilobase deletion including INO in this line. Ts and Bs lines were shown to have an identical deletion, indicating a common origin from a single deletion event. Analysis of microsatellite markers could not preclude the possibility that all three seedless accessions are vegetatively propagated clones. The sequence of the deletion site enabled a codominant assay for the wild-type and mutant genes allowing observation of complete cosegregation of the seedless/defective ovule phenotype with the INO deletion, showing maximal separation of less than 3.5 cM. The observed deletion is the only significant difference between the wild-type and Hs line over 587 kilobases, likely encompassing much more than 3.5 cM, showing that the deletion is the cause of seedless trait. The codominant markers and obtained progenies will be useful for introgression of the seedless trait into elite sugar apple lines and into other Annonas through interspecific crossings.
RESUMO
The generation of new hybrid varieties of tomato (Solanum lycopersicum L.) is the most widely used breeding method for this species and requires at least seven self-fertilization cycles to generate stable parent lines. The development of doubled haploids aims at obtaining completely homozygous lines in a single generation, although, to date, routine commercial application has not been possible in this species. In contrast, obtaining doubled haploid lines via gynogenesis has been successfully implemented in recalcitrant crops such as melon, cucumber, pumpkin, loquat and walnut. This review provides an overview of the requirements and advantages of gynogenesis as an inducer of haploidy in different agricultural crops, with the purpose of assessing the potential for its application in tomato breeding. Successful cases of gynogenesis variants involving in vitro culture of unfertilized ovules, use of 60Co-irradiated pollen, in vivo haploid inducers and wide hybridization are presented, suggesting that these methodologies could be implemented in tomato breeding programs to obtain doubled haploids.
RESUMO
Different breeding systems occur in the Gardenieae complex (Rubiaceae), from homoecy to dioecy which is present in two tribes, Gardenieae and Cordiereae. As part of a broad project focused on the reproductive anatomy of the species of these two tribes, we described the structural and functional differences of the gynoecium in the different floral morphs and determined the degree of gynoecium development in the staminate flowers. We conducted a comparative anatomical study focused on the gynoecium of one homoecious species (Tocoyena formosa, with perfect flowers) and three dioecious species (Genipa americana, Randia calycina, and Randia heteromera) of Gardenieae and one dioecious species (Cordiera concolor) of Cordiereae. The dioecious species have flowers that are morphologically perfect and functionally unisexual. Flowers in successive stages of development were collected, photographed, and fixed in formalin-acetic acid-alcohol. The material was examined using light microscopy and scanning electron microscopy. The anatomy of the ovary, style, and stigma was analyzed, and megasporogenesis and megagametogenesis were studied. The results achieved in this study, together with previously obtained data of the androecium, show that dioecy originated from homoecy in these species, since the unisexual flowers conserve some characteristics of the perfect flowers. In addition, a new type of ovule for the Rubiaceae family is described.
Assuntos
Rubiaceae , Flores/anatomia & histologia , Microscopia Eletrônica de Varredura , Óvulo Vegetal , Melhoramento VegetalRESUMO
PREMISE: Unlike most flowering plants, orchid flowers have under-developed ovules that complete development only after pollination. Classical studies reported variation in the stage in which ovule development is arrested, but the extent of this variation and its evolutionary and ecological significance are unclear. METHODS: Here, we used light microscopy to observe ovule development at anthesis for 39 species not previously studied and surveyed the literature gaining information on 94 orchid species. Tropical and temperate members of all five orchid subfamilies as well as species with contrasting pollination strategies (rewarding versus deceptive) and life forms (epiphytic versus terrestrial) were represented. We analyzed the data using statistical comparisons and a phylogenetic generalized least square (PGLS) analysis. RESULTS: Apostasioideae, the sister to the rest of the orchids, have mature ovules similar to other Asparagales, while under-differentiated ovules are present in the other subfamilies. Ovule developmental stages showed high variation even among closely related groups. Ovules were more developed in terrestrial than in epiphytic, in temperate than in tropical, and in rewarding than in deceptive pollination orchid species. This latter comparison was also significant in the PGLS analysis. CONCLUSIONS: These results suggest that ovule developmental stage in orchids can be shaped by ecological factors, such as seasonality and pollination strategy, and can be selected for optimizing female reproductive investment.
Assuntos
Orchidaceae , Óvulo Vegetal , Flores , Filogenia , PolinizaçãoRESUMO
Eragrostis curvula (Schrad.) Ness is a grass with a particular apomictic embryo sac development called Eragrostis type. Apomixis is a type of asexual reproduction that produces seeds without fertilization in which the resulting progeny is genetically identical to the mother plant and with the potential to fix the hybrid vigour from more than one generation, among other advantages. The absence of meiosis and the occurrence of only two rounds of mitosis instead of three during embryo sac development make this model unique and suitable to be transferred to economically important crops. Throughout this review, we highlight the advances in the knowledge of apomixis in E. curvula using different techniques such as cytoembryology, DNA methylation analyses, small-RNA-seq, RNA-seq, genome assembly, and genotyping by sequencing. The main bulk of evidence points out that apomixis is inherited as a single Mendelian factor, and it is regulated by genetic and epigenetic mechanisms controlled by a complex network. With all this information, we propose a model of the mechanisms involved in diplosporous apomixis in this grass. All the genetic and epigenetic resources generated in E. curvula to study the reproductive mode changed its status from an orphan to a well-characterised species.
RESUMO
Abstract Sweet cherry fruit is a tasty and valuable product for consumers. In order to increase the export share of cherry, which is also very important in export, it is beneficial to grow with cherry varieties that mature at different times. The cherries offered to the market in the early period will be more attractive. In this study, morphological and biological features of pistils of early-maturing 'Cristalina', 'Early Lory', 'Prime Giant', fruit set rates and pollen germination status and some chemical applications were investigated. As a result, fruit sets of cultivars were 17.6-28.6% in two years. Significant differences were observed in pistil morphology of the cultivars and 'Cristalina' had shorter pistil (14.35-14.51 mm) and style (11.47-11.65 mm) lengths than the other cultivars. Greater deformation was observed in primary ovules of 'Early Lory' right after anthesis. There were not significant differences in pollen germination ratios of the cultivars, but boric acid treatments improved pollen germination ratios of all cultivars. Boric acid application increased pollen germination with 21%. This was followed by IAA (8%), GA3 (5%), KNO3 (4%). It was concluded based on present findings that in orchard establishment with the early cultivars, flower biology should momentously be assessed.
Assuntos
Flores , Óvulo Vegetal , Prunus avium , PólenRESUMO
MAIN CONCLUSION: Characterization of anther and ovule developmental programs and expression analyses of stage-specific floral marker genes in Gossypium hirsutum allowed to build a comprehensive portrait of cotton flower development before fiber initiation. Gossypium hirsutum is the most important cotton species that is cultivated worldwide. Although cotton reproductive development is important for fiber production, since fiber is formed on the epidermis of mature ovules, cotton floral development remains poorly understood. Therefore, this work aims to characterize the cotton floral morphoanatomy by performing a detailed description of anther and ovule developmental programs and identifying stage-specific floral marker genes in G. hirsutum. Using light microscopy and scanning electron microscopy, we analyzed anther and ovule development during 11 stages of flower development. To better characterize the ovule development in cotton, we performed histochemical analyses to evaluate the accumulation of phenolic compounds, pectin, and sugar in ovule tissues. After identification of major hallmarks of floral development, three key stages were established in G. hirsutum floral development: in stage 1 (early-EF), sepal, petal, and stamen primordia were observed; in stage 2 (intermediate-IF), primordial ovules and anthers are present, and the differentiating archesporial cells were observed, marking the beginning of microsporogenesis; and in stage 6 (late-LF), flower buds presented initial anther tapetum degeneration and microspore were released from the tetrad, and nucellus and both inner and outer integuments are developing. We used transcriptome data of cotton EF, IF and LF stages to identify floral marker genes and evaluated their expression by real-time quantitative PCR (qPCR). Twelve marker genes were preferentially expressed in a stage-specific manner, including the putative homologs for AtLEAFY, AtAPETALA 3, AtAGAMOUS-LIKE 19 and AtMALE STERILITY 1, which are crucial for several aspects of reproductive development, such as flower organogenesis and anther and petal development. We also evaluated the expression profile of B-class MADS-box genes in G. hirsutum floral transcriptome (EF, IF, and LF). In addition, we performed a comparative analysis of developmental programs between Arabidopsis thaliana and G. hirsutum that considered major morphoanatomical and molecular processes of flower, anther, and ovule development. Our findings provide the first detailed analysis of cotton flower development.
Assuntos
Flores , Regulação da Expressão Gênica de Plantas , Gossypium , Flores/anatomia & histologia , Flores/genética , Perfilação da Expressão Gênica , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Óvulo Vegetal/genéticaRESUMO
Cassava (Manihot esculenta Crantz) is an important crop for subsistence farming in tropical and subtropical regions. There is a need to increase the rate of genetic gain to develop varieties adapted to new environmental conditions affected by climate change, which also influences the patterns of pests and diseases. The rate of cassava genetic improvement is limited by the difficulty in obtaining true-breeding types (inbred/homozygous lines). Cassava inbreeding obtained through conventional sequential self-pollination increases exposure of useful recessive traits and breeding value of progenitors. However, it takes 10-15 years to produce homozygous lines through successive self-pollination. Doubled haploid (DH) technology is a functional alternative to progressive self-pollination, and is already widely used in major crops to accelerate inbreeding. This work aimed at developing a protocol for the culture of isolated ovules and the induction of gynogenesis in cassava. Basic groundbreaking studies on cassava embryo sac development are presented. A protocol using unpollinated ovules collected from ovaries 1 day after anthesis is described. In the unpollinated-cultured ovules, the presence of embryos formed probably from the egg cells and not surrounded by the endosperm, was documented by anatomical analyses. This achievement is an important first step in the development of a reproducible gynogenesis protocol for the generation of doubled haploids in cassava. This protocol can also be useful as a starting point to obtain DHs using alternative methods of induction such as pollination of cassava with pollen of distant species or with cassava pollen irradiated with gamma rays.
RESUMO
This is the first study to describe in a timescale morphohistological and ultrastructural characteristics of fruit (cypsela) and seed development in Trichocline catharinensis, which was completed 21 days after anthesis (DAA). At anthesis, we identified an ovary with three differentiated regions, including the inner epidermis, inner part, and outer epidermis. The mature ovule showed an integument with the outer epidermis, integumentary parenchyma, and endothelium. Cells around the endothelium form the periendothelial zone with thick cell walls that showed Periodic acid-Schiff (PAS)-positive reaction. The periendothelial zone and endothelium showed degradation of the cells during embryogenesis. The main stages of embryo development from fecundation through mature seed were identified. The ripe cypsela showed the pericarp (exocarp), seed coat (exotesta), and remaining endosperm surrounding the embryo. Mature embryos were straight with shoot apical meristem (SAM), and root apical meristem (RAM) was separated by the hypocotyl. Light microscopy (LM) and transmission electron microscopy (TEM) analyses indicate cells with characteristics of meristem cells, as well as proteins and lipid bodies and mitochondria with few cristae in cotyledon cells. Our findings provide insight into taxonomic and physiological studies by detailing cypsela and seed ontogenesis from an endemic and vulnerable Asteraceae from southern Brazil. This study is also a starting point for establishing the biological criteria for seed harvesting and future studies of seed physiology and conservation of plant genetic resource.
Assuntos
Regulação da Expressão Gênica de Plantas/genética , Plantas/química , Sementes/química , BrasilRESUMO
Determining the in situ pattern of protein expression is crucial to accurately establish regulatory function and mode of action of any plant developmental program. Here, we describe two immunolocalization procedures that are consistently used to determine subcellular localization of ARGONAUTE proteins in the ovule of the Brassicaceae. The first is performed in resin-embedded semi-thin sections of developing ovules that can be observed under bright-field microscopy. The second is based in polyacrylamide immersion of complete (whole-mounted) gynoecia or ovules that are observed under confocal microscopy. Both procedures have been successfully performed to localize proteins involved in RNA-directed DNA methylation during the development of the anatropous bitegmic ovule in Arabidopsis, Brassica, or Boechera species.
Assuntos
Arabidopsis/genética , Proteínas Argonautas/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Óvulo Vegetal/genética , Proteínas de Arabidopsis/genética , Metilação de DNA/genética , Regulação da Expressão Gênica de Plantas/genéticaRESUMO
BACKGROUND: A classical dichotomous perspective proposes that either pollination or plant resources limit seed production. However, ovule number could also be limiting when pollination results in complete ovule fertilization and there are more plant resources available than needed to develop seeds. Moreover, this dichotomous view assumes that all flowers of a plant have equal access to a shared pool of resources, although these are frequently compartmentalized within plant modules, for example, inflorescences. How ovule number, pollination and resources affect seed production in physiologically-compartmentalized rather than physiologically-integrated plants has yet to be explored. We used raspberry (Rubus idaeus) to address this question. METHODS: We first assessed if ovule number affected the fraction of ovules that develop into seed (i.e., seed set) and whether this effect related to the extent of physiological integration among flowers within plants. This was achieved by statistically testing predictions on the sign and level of plant organization (i.e., among flowers within inflorescences, among inflorescences within ramets, and among ramets) of the relation between ovule number and seed set given different degrees of physiological integration. We then explored whether the relation between ovule number and seed set was affected by plant age (used here as a surrogate of resource availability) and pollination intensity (open-pollination vs. exclusion). RESULTS: Within inflorescences, flowers with more ovules set a larger fraction of seeds. On the other hand, seed set at the inflorescence level was negatively related to the average number of ovules per flower. Seed set increased with ovule number and open-pollination, and decreased with ramet age. However, ovule number explained more variation in seed set than ramet age and pollination treatment. Ramet age affected the strength of the relation of seed set to ovule number, which was stronger in old than young ramets. Pollination did not alter the strength of this relation to any significant extent. DISCUSSION: Results reveal the importance of ovule number as an overriding factor affecting seed set. Within inflorescences, resources appear to be differentially allocated to developing fruits from flowers with many ovules. This is consistent with the fact that in the raspberry a large proportion of the carbon invested in fruit development is fixed by the inflorescence subtending leaf. Differential resource allocation to flowers with many ovules is not affected by pollinator exclusion, being stronger in resource-exhausted ramets. This suggests that the effects of pollen limitation and resource allocation are compartmentalized at the inflorescence level. Consequently, modular plants can be viewed as reproductive mosaics where either ovule number, pollination or resources limit the number of seeds set by different flowers, so that improvements in any of them could increase plant seed production.
RESUMO
Ovule and seed development in plants has long fascinated the scientific community given the complex cell coordination implicated in these processes. These cell events are highly conserved but are not necessarily representative of all plants. In this study, with the aim of obtaining information regarding the cellular patterns that follow the usual development of the ovule and the zygotic embryo, we carried out an integral anatomical study of the Capsicum chinense Jacq., floral buds and seeds at various days during maturation. This study allowed us to identify the main histo-morphological stages accompanying the transition of somatic cells into the macrospore, female gamete, and the zygotic embryogenesis. This knowledge is fundamental for future biotechnological research focused on solving the morphological recalcitrance observed during the in vitro induction of somatic or microspore embryogenesis in Capsicum. For the first time in C. chinense, we have described the hypostases, a putative source of plant growth regulators, and "the corrosion cavity", a space around the embryo. Additionally, the cell wall pectin-esterification status was investigated by immunohistology. At early stages of morphogenesis, the pectin is highly methyl-esterified; however, methyl-esterification decreases gradually throughout the process. A comparison of the results obtained here, together with the histo- and immunological changes occurring during the somatic and microspore embryogenesis, should help to elucidate the biochemical mechanisms that trigger the morphogenic events in Capsicum spp.
Assuntos
Capsicum/crescimento & desenvolvimento , Óvulo Vegetal/crescimento & desenvolvimento , Pectinas/metabolismo , Sementes/crescimento & desenvolvimento , Capsicum/anatomia & histologia , Capsicum/metabolismo , Esterificação , Flores/anatomia & histologia , Flores/crescimento & desenvolvimento , Flores/metabolismo , Imunofluorescência , Óvulo Vegetal/anatomia & histologia , Óvulo Vegetal/metabolismo , Sementes/anatomia & histologia , Sementes/metabolismoRESUMO
KEY MESSAGE: BbrizGID1 is expressed in the nucellus of apomictic Brachiaria brizantha, previous to aposporous initial differentiation. AtGID1a overexpression triggers differentiation of Arabidopsis thaliana MMC-like cells, suggesting its involvement in ovule development. GIBBERELLIN-INSENSITIVE DWARF1 (GID1) is a gibberellin receptor previously identified in plants and associated with reproductive development, including ovule formation. In this work, we characterized the Brachiaria brizantha GID1 gene (BbrizGID1). BbrizGID1 showed up to 92% similarity to GID1-like gibberellin receptors of other plants of the Poaceae family and around 58% to GID1-like gibberellin receptors of Arabidopsis thaliana. BbrizGID1 was more expressed in ovaries at megasporogenesis than in ovaries at megagametogenesis of both sexual and apomictic plants. In ovules, BbrizGID1 transcripts were detected in the megaspore mother cell (MMC) of sexual and apomictic B. brizantha. Only in the apomictic plants, expression was also observed in the surrounding nucellar cells, a region in which aposporous initial cells differentiate to form the aposporic embryo sac. AtGID1a ectopic expression in Arabidopsis determines the formation of MMC-like cells in the nucellus, close to the MMC, that did not own MMC identity. Our results suggest that GID1 might be involved in the proper differentiation of a single MMC during ovule development and provide valuable information on the role of GID1 in sexual and apomictic reproduction.
Assuntos
Brachiaria/genética , Flores/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Óvulo Vegetal/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Apomixia/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Brachiaria/crescimento & desenvolvimento , Brachiaria/metabolismo , Flores/crescimento & desenvolvimento , Flores/metabolismo , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Poaceae/genética , Poaceae/crescimento & desenvolvimento , Poaceae/metabolismo , Pólen/genética , Pólen/crescimento & desenvolvimento , Pólen/metabolismo , Homologia de Sequência de AminoácidosRESUMO
In the majority of Latin-American countries, including Argentina, there is a limited availability of vaginal bioproducts containing probiotics in the market. In addition, the conventional treatments of genital tract infections in women represent a high cost to the public health systems. The future development of this type of bioproducts that employ specific lactobacilli strains would not only have a meaningful impact on women's health but would also represent a significant challenge to the pharmaceutical industry. The aims of the work described in this paper were (i) to study different pharmaceutical formulations of vaginal ovules containing Lactobacillus fermentum L23 and L. rhamnosus L60, to determine in which formulation lactobacilli viability was sustained for longer time and (ii) to evaluate if probiotic strains maintained both the antimicrobial activity and biofilm-producing ability after being recovered from the ovules. In this study, we developed and characterized three pharmaceutical formulations containing different glycerol amounts and specific lactobacilli strains. Three relevant parameters, cell viability, antimicrobial activity, and biofilm production, by lactobacilli recovered from the ovules were tested. Although the viability of L23 and L60 strains was mainly influenced by high ovule's glycerol proportion, they survived at 4 °C during the 180 days. Both lactobacilli's antimicrobial activity and biofilm-producing ability were maintained for all treatments. In conclusion, employing a much reduced number of components, we were able to select the most suitable pharmaceutical formulation which maintained not only lactobacilli viability for a long period of time but also their antimicrobial activity and biofilm-producing ability.