MIR822 modulates monosporic female gametogenesis through an ARGONAUTE9-dependent pathway in Arabidopsis thaliana

In the ovule of flowering plants, the establishment of the haploid generation occurs when a somatic cell differentiates into a Megaspore Mother Cell (MMC) and initiates meiosis. As most flowering plants, Arabidopsis thaliana undergoes a monosporic type of gametogenesis; three meiotically derived cells degenerate without further division, and a single one, the functional megaspore (FM), divides mitotically to form the female gametophyte. In Arabidopsis, the ARGONAUTE4 clade proteins are involved in the control of megasporogenesis. In particular, mutations in ARGONAUTE9 (AGO9) lead to the ectopic differentiation of gametic precursors that can give rise female gametophytes. However, the genetic basis and molecular mechanisms that control monosporic gametogenesis remain largely unknown. Here, we show that Arabidopsis plants carrying loss-of-function mutations in the AGO9-interacting miR822a give rise to extranumerary surviving megaspores that acquire a FM identity and divide without giving rise to differentiated female gametophytes. The overexpression of three miR822a target genes encoding Cysteine/Histidine-Rich C1 domain proteins (DC1) phenocopy mir822a plants. The miR822a targets are overexpressed in ago9 mutant ovules, confirming that miR822a acts through an AGO9-dependent pathway to negatively regulate DC1 domain proteins. Our results identify a new role of miRNAs in the most prevalent form of female gametogenesis in flowering plants

Developmental, ultrastructural and cytochemical investigations of the female gametophyte in Sedum rupestre L. (Crassulaceae)

This article describes the development of female gametophyte in Sedum rupestre L. New embryological information about the processes of megasporogenesis and megagametogenesis provided in this paper expand the current knowledge about the embryology of the studied species. S. rupestre is characterized by monosporic megasporogenesis and the formation of Polygonum–type embryo sac. The process of megasporogenesis is initiated by one megaspore mother cell, resulting in the formation of a triad of cells after meiosis and cytokinesis. The functional megaspore, which is located chalazally, is a mononuclear cell present next to the megaspore in the centre of the triad. Only one of the two non-functional cells of the triad is binucleate, which occur at the micropylar pole. In this paper, we explain the functional ultrastructure of the female gametophytic cells in S. rupestre. Initially, the cytoplasm of the gametophytic cells does not differ from each other; however, during differentiation, the cells reveal different morphologies. The antipodals and the synergids gradually become organelle-rich and metabolically active. The antipodal cells participate in the absorption and transport of nutrients from the nucellar cells towards the megagametophyte. Their ultrastructure shows the presence of plasmodesmata with electron-dense material, which is characteristic of Crassulaceae, and wall ingrowths in the outer walls. The ultrastructure of synergid cells is characterized by the presence of filiform apparatus and cytoplasm with active dictyosomes, abundant profiles of endoplasmic reticulum and numerous vesicles, which agrees with their main function—the secretion of pollen tube attractants. Reported data can be used to resolve the current taxonomic problems within the genus Sedum ser. Rupestria.

Gibberellin-mediated RGA-LIKE1 degradation regulates embryo sac development in Arabidopsis

Ovule development is essential for plant survival, as it allows correct embryo and seed development upon fertilization. The female gametophyte is formed in the central area of the nucellus during ovule development, in a complex developmental programme that involves key regulatory genes and the plant hormones auxins and brassinosteroids. Here we provide novel evidence of the role of gibberellins (GAs) in the control of megagametogenesis and embryo sac development, via the GA-dependent degradation of RGA-LIKE1 (RGL1) in the ovule primordia. YPet-rgl1Δ17 plants, which express a dominant version of RGL1, showed reduced fertility, mainly due to altered embryo sac formation that varied from partial to total ablation. YPet-rgl1Δ17 ovules followed normal development of the megaspore mother cell, meiosis, and formation of the functional megaspore, but YPet-rgl1Δ17 plants had impaired mitotic divisions of the functional megaspore. This phenotype is RGL1-specific, as it is not observed in any other dominant mutants of the DELLA proteins. Expression analysis of YPet-rgl1Δ17 coupled to in situ localization of bioactive GAs in ovule primordia led us to propose a mechanism of GA-mediated RGL1 degradation that allows proper embryo sac development. Taken together, our data unravel a novel specific role of GAs in the control of female gametophyte development.

Reproductive Multitasking: The Female Gametophyte

Fertilization of flowering plants requires the organization of complex tasks, many of which become integrated by the female gametophyte (FG). The FG is a few-celled haploid structure that orchestrates division of labor to coordinate successful interaction with the sperm cells and their transport vehicle, the pollen tube. As reproductive outcome is directly coupled to evolutionary success, the underlying mechanisms are under robust molecular control, including integrity check and repair mechanisms. Here, we review progress on understanding the development and function of the FG, starting with the functional megaspore, which represents the haploid founder cell of the FG. We highlight recent achievements that have greatly advanced our understanding of pollen tube attraction strategies and the mechanisms that regulate plant hybridization and gamete fusion. In addition, we discuss novel insights into plant polyploidization strategies that expand current concepts on the evolution of flowering plants.

Female gametogenesis and early seed development in Amaranthus hypochondriacus L.

<p><strong>Background</strong>: Attention to amaranth grains has increased in recent years due to the nutritional value of their seed proteins, which have high levels of the amino acid lysine. However, there is no detailed study describing the stages of seed development in <em>Amaranthus hypochondriacus. </em></p><p><strong>Question</strong>: How are the developmental patterns of the female gametophyte and young seed in <em>Amaranthus hypochondriacus</em>?</p><p><strong>Species studied</strong>: <em>Amaranthus hypochondriacus</em> L ’Revancha’ (Amaranthaceae).</p><p><strong>Study site and years of study</strong>: Plants were growth and collected from 2014 to 2016, in a greenhouse at Instituto Tecnológico de Celaya, Guanajuato, Mexico.</p><p><strong>Methods: </strong>Glomerules were collected before pollination and two weeks after anthesis. The ovules at different development stages were fixed and cleared and were analyzed by light microscopy. A clearing protocol was used to observe the developmental stages during female gametogenesis and embryogenesis.</p><p><strong>Results: </strong>We observed that the <em>Amaranthus hypochondriacus</em> ovule has a campylotropous form. The female gametophyte showed a<em> Polygonum</em>-type pattern of development. We were also able to identify all the stages from the megaspore mother cell to the cotyledon embryo stage. After meiosis, the micropylar megaspore differentiates into the functional megaspore. The embryo did not show symmetric divisions, although the final pattern is similar to that of in eudicotyledons. The suspensor showed additional longitudinal divisions, giving rise to a 2-rowed suspensor, while the endosperm showed a helobial development.</p><p><strong>Conclusions: </strong>These results will be used as baseline to identify morphological changes during seed development and to develop new strategies to improve seed quality or increase the yield.</p>

Cytological Study of Gender Conversion in Amur Grape

Amur grape (Vitis amurensis) is a dioecious species. To elucidate the time of and reason for pistil abortion in male amur grape from the perspective of cytology, we observed the sections of pistil of a male line during its development using optical and transmission electron microscopes. The abnormity in the morphology of nucellar cell and the development of various organelles appeared before the abnormity of functional megaspore mitosis. Programmed cell death (PCD) of the nucellar cells might be an important reason for mitosis disorder, leading to the abortion of pistil in male flower. However, the abortion can be eliminated by forchlorfenuron treatment, resulting in the recovery of functional pistil in male amur grape. This study provides cytological information on the gender conversion mechanism in male amur grape, which can promote gender determination studies in Vitis species.

Embriología de Pachycereus militaris (Audot) Hunt (Cactaceae)

This is a contribution to the embryology of cacti and to the definition of their reproductory structures. The development of anthers, ovules and seeds of Pachycereus militaris is described. The type of development of the anther wall is monocotyledonous. This may have taxonomic importance above the family level. The endothecium is formed by a single stratum and the pollen grains are tricolpate, spinulate and punctitegilate. A lineal triad of megaspores was observed. The functional megaspore is the chalazal one. It is proposed that the term campylotropous should be uti lized for describing the ovule type, while the term circinotropous should be reserved for the funicle. In contrast to the stated by other authors, this study suggests that the seeds of Pachycereus militaris should be considered as non-albuminous and non-perispermous.

Female sporogenesis in the native Antarctic grass Deschampsia antarctica Desv.

The development of megasporocytes and the functional megaspore formation in Deschampsia antarctica were analyzed with the use of microscopic methods. A single archesporial cell was formed directly under the epidermis in the micropylar region of the ovule without producing a parietal cell. In successive stages of development, the meiocyte was transformed into a megaspore tetrad after meiosis. Most megaspores were arranged in a linear fashion, but some tetrads were T-shaped. Only one of the 60 analyzed ovules contained a cell in the direct proximity of the megasporocyte, which could be an aposporous initial. Most of the evaluated D. antarctica ovules featured monosporic embryo sacs of the Polygonum type. Approximately 30% of ovules contained numerous megaspores that were enlarged. The megaspores were located at chalazal and micropylar poles, and some ovules featured two megaspores – terminal and medial – in the chalazal region, or even three megaspores at the chalazal pole. In those cases, the micropylar megaspore was significantly smaller than the remaining megaspores, and it did not have the characteristic features of functional megaspores. Meiocytes and megaspores of D. antarctica contained polysaccharides that were detectable by PAS-reaction and aniline blue staining. Starch granules and cell walls of megasporocytes, megaspores and nucellar cells were PAS-positive. Fluorescent callose deposits were identified in the micropylar end of the megasporocytes. During meiosis and after its completion, thick callose deposits were also visible in the periclinal walls and in a small amount in the anticlinal walls of megaspores forming linear and T-shaped tetrads. Callose deposits fluorescence was not observed in the walls of the nucellar cells.