Embryology in Helosis cayennensis (Balanophoraceae): Structure of Female Flowers, Fruit, Endosperm and Embryo

Helosis cayennensis (Balanophoraceae s.str.) is a holoparasite characterised by aberrant vegetative bodies and tiny, reduced unisexual flowers. Here, we analysed the development of female flowers to elucidate their morpho-anatomy and the historical controversy on embryo sac formation. We also studied the developmental origin of inflorescences and the ontogeny of fruits, embryo and endosperm and discussed in a phylogenetic framework. Inflorescences were analysed by optical, fluorescence and scanning electron microscopy. Inflorescences of H. cayennensis arise endogenously. Female flowers lack perianth organs, thus only consist of the ovary, two styles and stigmata. Ovules are undifferentiated; two megaspore mother cells develop inside a nucellar complex. The female gametophyte, named Helosis-type, is a bisporic four-celled embryo sac, provided with a typical egg apparatus and a uni-nucleated central cell. Fertilization was not observed, yet a few-celled embryo and cellular endosperm developed. In sum, results confirm that, among Santalales holoparasites, Helosis is intermediate in the reduction series of its floral organs. Although perianth absence best supports the Balanophoraceae s.str. clade, our literature survey on female flower developmental data across Balanophoraceae s.l. highlights the many gaps that need to be filled to really understand these features in the light of new phylogenetic relationships.

Early events during embryogenesis in Zea mays L.

The megagametophyte of <em>Zea mays</em> L. undergoes a series of structural changes after fertilization resulting in a well differentiated mature embryo and cellular endosperm at 480 hours after pollination in greenhouse conditions. In the present work emphasis was laid on the localization of the cytoplasm in the synergids, central cell-endosperm and egg cell-zygote prior to and after fertilization. The observations are discussed in relation to the process of early embryogenesis.

Light microscopical study of endosperm formation in Brassica napus L.

The cellularization of the endosperm of <em>Brassica napus</em> was investigated with light microscopy after embedment in Technovit 7100. The microtubular cytoskeleton was visualized by immunofluorescence techniques after embedment in butyl methyl metacrylate (BMM). The analyses of sectioned seeds, sampled at various developmental stages, revealed that the endosperm has a nuclear phase up to the early heart shaped stage of the embryo. From the heart shaped stage onwards cells and alveoli are formed in the endosperm. The cellularization of endosperm was preceded by alveolus formation except in the region surrounding the embryo suspensor and in the chalazal zone of the embryo sac where the nuclear endosperm vacuolated and cell walls were formed all around the nuclei. Alveolus formation only occurred when nuclei had attained a dense distribution in the layer of cytoplasm aligning the wall of the central cell. When nuclei divided within the alveoli, cell plate formation resulted in the formation of mononuclear endosperm cells along the embryo sac wall. When the walls of the alveoli grew towards the centre of the embryo sac, alveoli regularly closed leaving space for enlargement of the remaining alveoli. In this way endosperm cells enlarged going from the periphery to the central area of the embryo sac. The microtubular cytoskeleton was visualized in the nuclear, alveolar and cellular endosperm. The pattern observed, showed that the organization and function of the microtubular arrays was as generally found during endosperm development.

DNA endoreplication level in endosperm and in cotyledons during seed development in three dicotyledonous species

The DNA content of nuclear and cellular endosperm and in cotyledons in the course of seed development in <i>Helianthus annuus</i>, <i>Pisum sativum</i> and <i>Vicia faba</i> was cytophotometrically measured after Faulgen's reaction. <i>Helianthus annuus</i> is characterized by the lowest endoreplication dynamics attaining a maximum DNA content of 48C in nuclear endosperm, which corresponds to four endoreplication rounds, and 16C DNA (three rounds) in cotyledons. In <i>Pisum sativum</i> cellular endosperm in the studied stages of seed development was absent and the highest DNA content in cotyledons was 512C DNA; this corresponds to eight endoreplication rounds. In <i>Vicia faba</i>, the maximum DNA endoreplication level was 1536C in nuclear endosperm, which corresponds to nine endoreplication rounds; in cotyledons the maximum DNA content was 64C (five rounds). In the course of seed development, differences in the dynamics of DNA endoreplication in throphic tissues of the examined species seem to be similar to differences in the endoreplication level in root and leaf parenchyma, but the DNA endoreplication level is much higher in throphic tissues. Therefore, the endoreplication pattern might be species-specific and genetically conditioned.

Dynamics of early embryo development in reference to endosperm and embryo types in angiosperms

The mitotic cycle of the endosperm cells is relatively short in the firsit phases of the postfertilization development. The endosperm type does not significantly influence the duration of the mitotic cycle; it might, however, influence the dynamics of zygote and embryo development. The quick development and early end of cellular endosperm proliferation is connected with the fact that it is, in most cases, bound to small, "spare" or "saving" tenuinucellate and unitegmic ovules. Structural differences in the behaviour of the endosperm of different types, in the phase of globular and early heart embryo, might point to differences in the time or way of transition of the embryo from suspensorial to surface nutrition.

Morphology of Postpollination Fruit Abortion in Pecan

Anatomy of normal and abortive fruit was compared at each of the three postpollination fruit drops characteristic of pecan [Carya illinoensis (Wangenh.) C. Koch]. Size differences between normal and abortive fruit decreased during the growing season, but differences in ovule size between normal and abortive fruit increased. During Drop II, normal and abortive fruit had an integument enclosing a massive nucellus in which an embryo sac was embedded, but embryo sac shape and constituents differed. Embryo sacs were distended in normal fruit and contained a definitive zygote as evidence of fertilization, i.e., union of egg and sperm. In contrast, embryo sacs in abortive fruit were shriveled and contained an egg apparatus as in unfertilized distillate flowers. During Drop III, normal and abortive fruit had a similar multicellular embryo. The nucellus in normal fruit was reduced to a cap at the micropyle region and cellular endosperm was evident. In contrast, the nucellus in abortive fruit was abundant and cellular endosperm was not evident. During Drop IV, embryo development in abortive fruit lagged behind that of normal fruit. Thus, we present the first direct evidence that aborted pecans deviate from normal fruit by an absence of a zygote at Drop II, a deficiency in cellular endosperm at Drop III, and a delay in embryo development at Drop IV.

Microtubular configurations during endosperm development in Phaseolus vulgaris

Microtubular cytoskeletons in nuclear, alveolar, and cellular endosperm of bean (Phaseolus vulgaris) were analyzed immunocytochemically and by electron microscopy to reveal their function during cellularization. Nuclear endosperm showed a fine network of microtubules between the wide-spaced nuclei observed towards the chalazal pole. Near the embryo, where nuclei were densely packed, bundles of microtubules radiated from nuclei. They were formed just before alveolus formation and functioned in spacing nuclei and in forming internuclear, phragmoplast-like structures that gave rise to nonmitosis-related cell plates. During alveolus formation cell plates extended and fused with other newly formed walls, thus forming the walls of alveoli. Growing wall edges of cell plates exhibited arrays of microtubules perpendicular to the plane of the wall, initially. When two growing walls were about to fuse, microtubules of both walls interacted, and because of the interaction of microtubules, the cell walls changed their position. When a growing wall was about to fuse with an already existing wall, such interactions between microtubules were not observed. It is therefore concluded that interactions of microtubules of fusing walls influence shape and position of walls. Thus microtubules control the dynamics of cell wall positioning and initial cell shaping. Key words: cell wall, cellularization, endosperm, microtubule, Phaseolus vulgaris.

Cellular endosperm formation in Phaseolus vulgaris. I. Light and scanning electron microscopy

The formation of the endosperm in Phaseolus vulgaris L. conforms to the nuclear pattern of endosperm development. The endosperm is partially cellularized in the vicinity of the developing embryo, while the rest of the endosperm remains multinucleate. Mitotic activity of the endosperm is gradually confined to the region adjacent to the tips of the enlarging cotyledons. Continuing mitotic activity in this region results in the formation of cellular endosperm in the bean seed. At the cotyledon stage of embryo development, except in the region of the degenerating nucellus, the entire surface of the developing embryo is covered by a layer of cellular endosperm cells. Initially, the cellular endosperm is loosely attached to the inner surface of the seed coat. With the disappearance of the liquid endosperm, it becomes firmly attached. Further expansion of the seed results in the separation of cellular endosperm cells along their long axes. As the seed matures, the cellular endosperm dries, with no apparent degradation of its cells.