Endosperm development in Dyckia pseudococcinea (Pitcairnioideae - Bromeliaceae)

Dyckia pseudococcinea is a threatened species endemic to the restingas of the Atlantic Forest, an area under strong anthropic impact. From the perspective of conservation, plant embryology is interesting by the variety of approaches for in vitro culture of seedlings. With this in mind, the present study describes the endospermogenesis of D. pseudococcinea to provide data for conservation studies and indicate characters that may contribute to the systematics of Pitcairnioideae. Using anatomical analysis, samples of flowers and fruits at different developmental stages were analyzed. We observed the establishment of the primary endosperm nucleus and then the primary endosperm cell (PEC). Upon expansion of the central vacuole of PEC, nuclei migrate to the chalazal, peripheral, and micropylar domains. An early centripetal cellularization begins in the chalazal region, characterizing the endosperm as coenocytic/multicellular type. With cellularization, the endosperm enfolds the embryo. In addition, in the outermost layer of the endosperm, an aleurone layer is visible. These observations allowed some embryological characters to be identified, thus helping to clarify the systematic relationships of Pitcairnioideae and also the genus Dyckia, such as the presence of hypostasis, the persistence of the antipodals and synergids during the early stages of endospermogenesis and coenocytic/multicellular endospermogenesis. Having established the details of endospermogenesis, we then set forth guidelines for the development of in vitro culture protocols aimed at the conservation of D. pseudococcinea, an endangered Bromeliad species of the Atlantic Forest.

Fertilisation of polar nuclei and formation of early endosperms in Dendrobium catenatum: evidence for the second fertilisation in Orchidaceae

Whether the second fertilisation, i.e. fertilisation of polar nuclei, or fusion of the second sperm with polar nuclei occurs in Orchidaceae has long been controversial because of lack of evidence. In the present study, we observed fusion and fertilisation of polar nuclei and formation of early endosperms in the orchid Dendrobium catenatum Lindl., by using a resin-embedded section technique. As the product of the second fertilisation, the primary endosperm nucleus (fertilised polar nuclei) can last until the global embryo stage, indicating that initiation of endosperm development and that of embryo development were fully asynchronous. The present study demonstrated the occurrence of the second fertilisation in D. catenatum by providing lines of new evidence.

Pollen Tube Growth and Double Fertilization in Camellia oleifera

Camellia oleifera is an important plant species that produces edible oils. Understanding the double fertilization of this plant is critical for studies concerning crossbreeding, self-incompatibility, and the biological mechanisms underlying hybridization. We aimed to characterize pollen tube growth and double fertilization in C. oleifera. The female and male parent cultivars (Huashuo and Xianglin XLC15, respectively) were used for artificial pollination. Growth of the pollen tube in the style, ovary, and ovule from pollination to fertilization and the cytological characteristics of female and male gamete fusion during double fertilization were observed using fluorescence and scanning electron microscopy (SEM). Numerous pollen grains germinated 2 to 4 hours after pollination. The pollen tubes entered the interspaces between the papillar cells, grew along the stylar canal, and aggregated at the one-third site of the style. They grew in the gradually narrowing stylar canal, entering the locule. The tubes turned 90° and entered the embryo sac through the micropyle; subsequently, they entered a degenerated synergid, where the spermatids were released. One sperm nucleus fused with the polar nucleus, forming the primary endosperm nucleus, whereas the other sperm fused with the egg, forming the zygote. The polar nucleus was fertilized earlier than the egg. Double fertilization of C. oleifera is characterized as pre-mitotic gametogony. The current results lay a theoretical foundation for studies concerning the crossbreeding and embryology of C. oleifera and provide fundamental data concerning the reproductive biology of the genus Camellia.

Embryo and endosperm development in coriander (Coriandrum sativum)

Coriander ( Coriandrum sativum L.) seeds are rich in lipids and are potentially important sources of oils for industrial use. The objective of this study was to determine the details of embryo and endosperm development and the sites of storage reserves using microscopy and histochemistry. In coriander, the zygote divides unequally, giving rise to a large basal cell and a smaller terminal cell. Subsequent divisions in the basal cell result in the formation of a suspensor, and divisions in the terminal cell give rise to cells of the embryo proper. A defined cell lineage is absent in the formation of the proembryo. Contrary to other flowering plants, the suspensor persists as the embryo matures and storage products are present within the cytoplasm of the suspensor cells. After fertilization, the primary endosperm nucleus divides rapidly, resulting in a large syncytium of nuclei and cytoplasm. The rapid nuclear divisions occur prior to the first division of the zygote. Cellularization of the endosperm occurs soon after. Within the developing seed, the endosperm can be separated into two main regions, i.e., the “embryo surround region” (ESR) of endosperm and the persistent endosperm. The endosperm cells in these two regions have different cell fates and storage products. In the ESR, the endosperm cells undergo distinct structural changes and are destined to degenerate. These endosperm cells produce a significant amount of polysaccharides and these materials appear to aid in cell separation prior to cell degeneration. At the boundary of the ESR, the endosperm cells are partially degenerated with a large accumulation of lipids. The bulk of the endosperm cells next to the seed coat persist and they are responsible for the production and accumulation of storage lipids and proteins.

Structural and ultrastructural characterization of maize coenocyte and endosperm cellularization

Maize coenocytic and cellularizing endosperm development were characterized at optical and transmission electron microscopy levels. Samples were collected daily in 3 consecutive years under different temperature regimes; therefore, the developmental stages were expressed on a growing degree basis. Soon after the primary endosperm nucleus is formed, it starts dividing without cytokinesis, leading to the formation of the coenocyte. The nuclei divide freely on the periphery of the coenocyte and spread from the micropylar region toward the chalazal region. The first anticlinal walls are formed as the nuclei come closer to each other, followed by periclinal wall formation also starting in the micropylar region. As cellularization proceeds, new anticlinal and periclinal walls assume a less organized pattern until the central vacuole is fully occupied by newly formed cells. Overall, our findings suggest that the developmental pattern of maize coenocytic and cellularizing endosperm has many similarities to other species of the Poaceae family.

Ultrastructure of the fertilized embryo sac in the dwarf mistletoe Arceuthobium americanum (Viscaceae) and development of the caecum

Post-fertilization changes in the seven-celled embryo sac of the parasitic angiosperm Arceuthobium americanum Nutt. ex Engelm. were investigated with light, fluorescence, and electron microscopy. Two embryo sacs, of which only one is fertilized, are found within the reduced ategmic ovule (the placental–nucellar complex). All cells of the fertilized embryo sac developed thickened cellulosic and (or) hemicellulosic bounding walls and, aside from the degenerative synergid, acquired ultrastructure reflective of metabolically active cells. The filiform apparatus became thickened in the persistent synergid, and a similar structure developed in the antipodals. Unlike the changes observed in the zygote and central cell, those in the persistent synergid and antipodals were unusual, as these cells regularly degenerate after fertilization in a typical angiosperm; therefore, in A. americanum, they likely play a role in providing nutrition to the zygote and (or) central cell. A pouch-like outgrowth (caecum) initiated from the central cell near the primary endosperm nucleus and, after vacuolar expansion and intercellular growth, reached the base of the placental–nucellar complex in three days. The observation of mitochondria clusters within the central cell, caecum, and adjacent placental–nucellar complex cells suggests the caecum functions in embryo sac expansion and haustorial nutrient absorption.Key words: Arceuthobium, caecum, cell walls, fertilized embryo sac, mistletoe, ultrastructure.

Apomixis in flowering plants: an overview

Apomixis is a common feature of perennial plants, which occurs in ca . 60% of the British flora, but has been largely ignored by reproductive theoreticians. Successful individuals may cover huge areas, and live to great ages, favoured by ‘symmetrical’ selection. Apomixis is favoured by colonizing modes, for instance post–glacially. Despite its theoretical advantages, apomixis usually coexists with sexuality, suggesting ‘hidden’ disadvantages. Agamospermy (apomixis by seed) is relatively uncommon, but gains from the attributes of the seed. It pays agamospermy genes, which discourage recombination, to form co–adapted linkage groups, so that they become targets for disadvantageous recessive mutant accumulation. Consequently, agamospermy genes cannot succeed in diploids and agamosperms are hybrid and highly heterotic. Agamospermous endosperm may suffer from genomic imbalance, so that nutritious ovules, which can support embryos without endosperm, may be preadapted for agamospermy. When primary endosperm nucleus fertilization (‘pseudogamy’) continues as a requirement for many aposporous agamosperms, selfing sex becomes preadaptive and archesporial sex remains an option. Apomictic populations can be quite variable although apomictic families are much less variable than sexuals. Only in some diplosporous species does sex disappear completely, and in those species some release of variability may persist through somatic recombination. The search for an agamospermy gene suitable for genetic modification should target fertile sexuals with a single localized agamospermy ( A ) gene, which therefore lack a genetic load. The A gene should coexist alongside sexuality, so that it would be easy to select seedlings of sexual and asexual origins. Plants with sporophytic agamospermy provide all these attributes.

LIGHT AND ELECTRON MICROSCOPIC EXAMINATION OF THE EMBRYO AND ENDOSPERM DEVELOPMENT IN THE NATURAL TETRAPLOID TRIFOLIUM PRATENSE L.

The ultrastructure of the embryo cells in ovules, from fertilization to the embryo maturity stage in the natural tetraploid Trifolium pratense L. that has a very low rate of seed formation, was examined. Following fertilization the vacuolar organization in the zygote changes. The zygote was a polarized cell and contained a central nucleus, mitochondria, plastids, ribosomes. and lipid bodies. Ribosomal concentration increases significantly after fertilization. The first division of the zygote was transverse or oblique and unequal. The primary endosperm nucleus divides before the zygote nucleus, forming a coenocytic nuclear endosperm; however, part of it later becomes cellular. At the earliest stage of embryo development, the cells were vacuolate, and plastids and mitochondria were simple in structure. During all stages of embryogenesis the suspensor cells were less electron dense than the adjoining embryo cells. Endosperm cellularization begins when the embryo has developed the globular embryo proper. Cellularization starts at the micropylar end of the embryo sac and progresses toward the chalazal end. Dictyosome activity, ribosomal aggregation, and the amount of rough endoplasmic reticulum were highest during the late globular embryo stage. In addition, the vacuolar volume in the cells was reduced. Lipid bodies were present up to the early globular stage, then disappeared. The inner cell walls of the embryo were thin, with many plasmodesmata. These walls begin to thicken at the late globular stage. The results show a corresponding increase in the amount and activity of the metabolic machinery as the development of the embryo progresses.

Early events in the embryo sac after intraspecific and interspecific pollinations in Rhododendron kawakamii and R. retusum

Early events in the embryo sac of Rhododendron kawakamii and R. retusum have been studied after compatible self-pollinations and eight interspecific crosses, using sectioned ovaries, pistil squashes, and seed-set data. Ovules of Rhododendron kawakamii and R. retusum are anatropous, unitegmic, and tenuinucellate, with a typical eight-nucleate, seven-celled embryo sac. Fertilization normally occurs 4–5 days after pollination. The zygote lays down a callose wall but remains undivided during the first 13–15 days after pollination. The primary endosperm nucleus divides soon after fertilization, and development is cellular ab initio. Crosses of R. kawakamii (♂) with R. santapaui and R. retusum and crosses of R. retusum (♂) with R. kawakamii, R. santapaui, R. ovatum, and R. tashiroi showed apparently normal fertilization in a majority of ovules entered by pollen tubes. In crosses of R. kawakamii (♂) with R. quadrasianum and Kalmia latifolia entry of pollen tubes into ovules was delayed and frequently abnormal. Apart from compatible self-pollinations of R. kawakamii an R. retusum, only the cross of R. kawakamii (♂) with R. santapaui produced healthy seedlings. Of the remaining seven interspecific crosses only three showed significant embryo development in control pistils left to mature in situ. Similarities and differences in the breeding behaviour of R. kawakamii and R. retusum are discussed with reference to their taxonomic grouping within subsection Pseudovireya.