Seed development in Paeonia ostii (Paeoniaceae), with particular reference to embryogeny

Background Seeds of Paeonia ostii have been proposed as a source of raw material for the production of edible oil; however, lack of information about the developmental biology of the seeds hampers our ability to use them. Our aim was to investigate development of the seed coat, endosperm and embryo of P. ostii in relation to timing of accumulation of nutrient reserves from pollination to seed maturity. Ovules and developing seeds of P. ostii were collected at various stages of development from zygote to maturity. Seed fresh mass, dry mass, germination, moisture, soluble sugars, starch, protein and oil content were determined. Ontogeny of seeds including embryo, endosperm and seed coat were analyzed histologically. Results The ovule of P. ostii is anatropous, crassinucellate and bitegmic. The zygote begins to divide at about 5 days after pollination (DAP), and the division is not accompanied by cell wall formation. By 25 DAP, the proembryo begins to cellularize. Thereafter, several embryo primordia appear at the surface of the cellularized proembryo, but only one matures. Endosperm development follows the typical nuclear type. The seed coat is derived from the outer integument. During seed development, soluble sugars, starch and crude fat content increased and then decreased, with maximum contents at 60, 80 and 100 DAP, respectively. Protein content was relatively low compared with soluble sugars and crude fat, but it increased throughout seed development. Conclusions During seed development in P. ostii, the seed coat acts as a temporary storage tissue. Embryo development of P. ostii can be divided into two stages: a coenocytic proembryo from zygote (n + n) that degenerates and a somatic embryo from peripheral cells of the proembryo (2n → 2n). This pattern of embryogeny differs from that of all other angiosperms, but it is similar to that of gymnosperms.

4-CPA (4-Chlorophenoxyacetic Acid) Induces the Formation and Development of Defective “Fenghou” (Vitis vinifera × V. labrusca) Grape Seeds

For some horticultural plants, auxins can not only induce normal fruit setting but also form fake seeds in the induced fruits. This phenomenon is relatively rare, and, so far, the underlying mechanism remains unclear. In this study, “Fenghou” (Vitis vinifera × V. labrusca) grapes were artificially emasculated before flowering and then sprayed with 4-CPA (4-chlorophenoxyacetic acid) to analyze its effect on seed formation. The results show that 4-CPA can induce normal fruit setting in “Fenghou” grapes. Although more seeds were detected in the fruits of the 4-CPA-treated grapevine, most seeds were immature. There was no significant difference in the seed shape; namely, both fruit seeds of the grapevines with and without 4-CPA treatment contained a hard seed coat. However, the immature seeds lacked embryo and endosperm tissue and could not germinate successfully; these were considered defective seeds. Tissue structure observation of defective seeds revealed that a lot of tissue redifferentiation occurred at the top of the ovule, which increased the number of cell layers of the outer integument; some even differentiated into new ovule primordia. The qRT-PCR results demonstrated that 4-CPA application regulated the expression of the genes VvARF2 and VvAP2, which are associated with integument development in “Fenghou” grape ovules. Together, this study evokes the regulatory role of 4-CPA in the division and continuous redifferentiation of integument cells, which eventually develop into defective seeds with thick seed coats in grapes.

Functional conservation of the grapevine candidate gene INNER NO OUTER for ovule development and seed formation

Seedlessness represents a highly appreciated trait in table grapes. Based on an interesting case of seedless fruit production described in the crop species Annona squamosa, we focused on the Vitis vinifera INNER NO OUTER (INO) gene as a candidate. This gene encodes a transcription factor belonging to the YABBY family involved in the determination of abaxial identity in several organs. In Arabidopsis thaliana, this gene was shown to be essential for the formation and asymmetric growth of the ovule outer integument and its mutation leads to a phenotypic defect of ovules and failure in seed formation. In this study, we identified in silico the V. vinifera orthologue and investigated its phylogenetic relationship to INO genes from other species and its expression in different organs in seeded and seedless varieties. Applying cross-species complementation, we have tested its functionality in the Arabidopsis ino-1 mutant. We show that the V. vinifera INO successfully rescues the ovule outer integument growth and seeds set and also partially complements the outer integument asymmetric growth in the Arabidopsis mutant, differently from orthologues from other species. These data demonstrate that VviINO retains similar activity and protein targets in grapevine as in Arabidopsis. Potential implications for grapevine breeding are discussed.

Embryology of Styracaceae and implications for the evolution of the integument number in Ericales

Phylogenetic relationships in Styracaceae are well understood, but embryological characters and the ontogeny of integument(s) are still uncertain in many species. The goals of this study are to evaluate the systematic implications of embryological characters in Styracaceae, clarify the character evolution of the number of integuments and suggest a mechanism for the transition between unitegmic and bitegmic ovules. We examined the embryological characters of four genera and five species of Styracaceae, most of which were shared across taxa. However, Styrax has specific embryological features including bitegmic ovules, a multiplicative and sclerotic outer mesotesta and vascular bundles in the testa, all possible autapomorphies. The other three genera of Styracaceae share a unitegmic ovule, a parenchymatous mesotesta and a seed coat without vascular bundles, possible plesiomorphies with Diapensiaceae and Symplocaceae. The transition from a unitegmic to a bitegmic condition can be interpreted to be caused by a downwards shift of the boundary between the inner and outer integument, due to reduced activity in the subdermal initials and increased activity in the dermal initials of the outer integument at its base.

Seed Biology of Lepidium apetalum (Brassicaceae), with Particular Reference to Dormancy and Mucilage Development

Lepidium apetalum (Brassicaceae) is an annual or biennial weed widely distributed in Asia and Europe. The outer surface of L. apetalum seeds produces a large amount of mucilage. The primary aim of this study was to explore the dormancy characteristics and to determine how mucilage develops. The role of mucilage in water absorption/dehydration, the effects of after-ripening, gibberellin acid (GA3), cold stratification and seed coat scarification on germination, the role of mucilage in germination and seedling growth during drought, and the progress of mucilage production during seed development were investigated. The results indicate that the best temperature regime for germination was 10/20 °C. After-ripening, GA3 and seed coat scarification helped to break dormancy. Light promoted germination. Seedling growth of mucilaged seeds were significantly higher than those of demucilaged seeds at −0.606 and −1.027 MPa. Anatomical changes during seed development showed that mucilage was derived from the outer layer of the outer integument cells. Our findings suggest that seeds of L. apetalum exhibited non-deep physiological dormancy. The dormancy characteristics along with mucilage production give seeds of L. apetalum a competitive advantage over other species, and thus contribute to its potential as a weed. Effective control of this weed can be achieved by deep tillage.

Seed coat development in explosively dispersed seeds of Cardamine hirsuta

Background and Aims Seeds are dispersed by explosive coiling of the fruit valves in Cardamine hirsuta. This rapid coiling launches the small seeds on ballistic trajectories to spread over a 2 m radius around the parent plant. The seed surface interacts with both the coiling fruit valve during launch and subsequently with the air during flight. We aim to identify features of the seed surface that may contribute to these interactions by characterizing seed coat differentiation. Methods Differentiation of the outermost seed coat layers from the outer integuments of the ovule involves dramatic cellular changes that we characterize in detail at the light and electron microscopical level including immunofluorescence and immunogold labelling. Key Results We found that the two outer integument (oi) layers of the seed coat contributed differently to the topography of the seed surface in the explosively dispersed seeds of C. hirsuta vs. the related species Arabidopsis thaliana where seed dispersal is non-explosive. The surface of A. thaliana seeds is shaped by the columella and the anticlinal cell walls of the epidermal oi2 layer. In contrast, the surface of C. hirsuta seeds is shaped by a network of prominent ridges formed by the anticlinal walls of asymmetrically thickened cells of the sub-epidermal oi1 layer, especially at the seed margin. Both the oi2 and oi1 cell layers in C. hirsuta seeds are characterized by specialized, pectin-rich cell walls that are deposited asymmetrically in the cell. Conclusions The two outermost seed coat layers in C. hirsuta have distinct properties: the sub-epidermal oi1 layer determines the topography of the seed surface, while the epidermal oi2 layer accumulates mucilage. These properties are influenced by polar deposition of distinct pectin polysaccharides in the cell wall. Although the ridged seed surface formed by oi1 cell walls is associated with ballistic dispersal in C. hirsuta, it is not restricted to explosively dispersed seeds in the Brassicaceae.

Megasporogenesis, Microsporogenesis, and Development of Female and Male Gametophytes of Ziziphus jujuba Mill. ‘Zhongqiusucui’

To investigate whether reproductive disorders exist in the sexual reproduction of Ziziphus jujuba Mill. ‘Zhongqiusucui’ and to understand the reproductive biology of ‘Zhongqiusucui’ and genetic improvements in jujube trees, we used ‘Zhongqiusucui’ flowers at different developmental stages as materials and conducted field and microscopic observations on the developmental pattern of mega- and microsporogenesis, as well as on the development of male and female gametophytes. The results show the following. 1) From the inflorescence development stage to flowering, the grade 0 bud on the inflorescence exhibited an increase in horizontal diameter, longitudinal diameter, peduncle length, and bud weight, but the rates of increase were different. From day 1 to day 5 after the inflorescence had developed, floral buds mostly grew horizontally. Day 5 was the floral bud flattening stage. From day 6 to day 8 after the inflorescence had developed, floral buds mostly grew longitudinally, and day 8 was the floral bud enlarging stage. 2) The stamens of ‘Zhongqiusucui’ had five anthers, and there were four locules per anther. The anther wall consisted of epidermis, endothecium, one- to two-layered middle layer, and a secretory-type tapetum. In addition, the development of the anther wall belonged to the basic type. The cytokinesis of the microsporocytes was synchronous, the tetrads mostly arranged as a tetrahedron, and the mature pollen had three germ pores, three grooves, and was bicellular pollen. During meiosis, the microsporocytes in each locule were at the same phase and therefore exhibited synchrony. Among the different anthers in the same floral bud, as well as the four locules in the same anther, the microsporocytes had asynchronous meiosis. 3) The pistils in the ‘Zhongqiusucui’ had two ovaries, two anatropous ovules, inner and outer integument, crassinucellate tetrads formed by the meiosis of megasporocytes aligned linearly along the nucellus, megaspore at the chalazal end that developed into the functional megaspore, which underwent mitotic division three times and developed into the mature embryo sac containing seven cells and eight nuclei, and embryo sac development of the Polygonum type. 4) The external morphology of the ‘Zhongqiusucui’ floral buds correlated with the internal developmental stage of the male and female gametophyte. Therefore, the internal developmental progress of the stamen and pistil can be determined by the external morphological characteristics of the floral buds.

Floral organogenesis of Prunus laurocerasus and P. serotina and its significance for the systematics of the genus and androecium diversity in Rosaceae

Phylogenetic studies have shown that most clades in Prunus are well-supported by the flower structure, but most taxa in the racemose group have not yet been re-evaluated and could contribute to the understanding of the systematic relationships of the subgenera. We examined the inflorescence and flower development in Prunus laurocerasus L. (subgenus Laurocerasus) and P. serotina Ehrh. (subgenus Padus I) using scanning electron microscopy. Our results indicate that they share several floral development characters but differ in the following aspects: (i) all of their flowers are fully developed and each flower is enclosed by a bract and two bracteoles, which later stop development (vs. the terminal flower degenerates and only a single bract subtends each flower); (ii) the style protrudes from the floral bud (vs. the style is crooked and below the anthers); (iii) the outer integument initiates close to the inner one (vs. in the middle of the ovule); and (iv) an obturator appears after initiation of the two integuments (vs. simultaneously with the inner integument). Although our results are preliminary, differences in floral developmental characters support the different origins of Prunus subgenera Laurocerasus and Padus as based on molecular phylogenetic studies.

‘Sensory pad’- A novel chemoreceptive device in Hilsa (Tenualosa ilisha) to support its amphihaline attribute

Hilsa, Tenualosa ilisha is an amphihaline migratory fish that performs spawning migration to selected freshwater rivers in Indo-Pacific region. It is not clear what force triggers its migration. In this paper, we attempted to describe the features of outer integument from its head region as chemosensory site which appears to play significant role in its upstream migration. We found that this area (termed as snout) has very soft and scale less tissue oriented with pit like grooves named as ‘epidermal pit’. Around these pits, odorant receptor G-protein subunits (Gαq, Gαs/olf and Gαo) have been substantially localized. Use of DASPEI also traced this area with neuronal existence. These features in the snout likely to contribute for chemosensory requirements of the fish during upstream migration. Considering such findings, we named this area of snout as ‘sensory pad’. Its position at the forefront of olfactory organ and brain may have important role in facilitating sensory reception by the fish swimming upstream to the river.