Plant egg cell fate determination depends on its exact position in female gametophyte

Plant fertilization involves both an egg cell, which fuses with a sperm cell, and synergid cells, which guide pollen tubes for sperm cell delivery. Therefore, egg and synergid cell functional specifications are prerequisites for successful fertilization. However, how the egg and synergid cells, referred to as the “egg apparatus,” derived from one mother cell develop into distinct cell types remains an unanswered question. In this report, we show that the final position of the nuclei in female gametophyte determines the cell fate of the egg apparatus. We established a live imaging system to visualize the dynamics of nuclear positioning and cell identity establishment in the female gametophyte. We observed that free nuclei should migrate to a specific position before egg apparatus specialization. Artificial changing in the nuclear position on disturbance of the actin cytoskeleton, either in vitro or in vivo, could reset the cell fate of the egg apparatus. We also found that nuclei of the same origin moved to different positions and then showed different cell identities, whereas nuclei of different origins moved to the same position showed the same cell identity, indicating that the final positions of the nuclei, rather than specific nucleus lineage, play critical roles in the egg apparatus specification. Furthermore, the active auxin level was higher in the egg cell than in synergid cells. Auxin transport inhibitor could decrease the auxin level in egg cells and impair egg cell identity, suggesting that directional and accurate auxin distribution likely acts as a positional cue for egg apparatus specialization.

Ovary Signals for Pollen Tube Guidance in Chalazogamous Mangifera indica L.

Most flowering plants show porogamy in which the pollen tubes reach the egg apparatus through the micropyle. However, several species show chalazogamy, an unusual pollen tube growth, in which the pollen tubes reach the embryo sac through the chalaza. While ovary signals for pollen tube growth and guidance have been extensively studied in porogamous species, few studies have addressed the process in chalazogamous species such as mango (Mangifera indica L.), one of the five most important fruit crops worldwide in terms of production. In this study, we characterize pollen–pistil interaction in mango, paying special attention to three key players known to be involved in the directional pollen tube growth of porogamous species such as starch, arabinogalactan proteins (AGPs), and γ-aminobutyric acid (GABA). Starch grains were observed in the style and in the ponticulus at anthesis, but their number decreased 1 day after anthesis. AGPs, revealed by JIM8 and JIM13 antibodies, were homogenously observed in the style and ovary, but were more conspicuous in the nucellus around the egg apparatus. GABA, revealed by anti-GABA antibodies, was specifically observed in the transmitting tissue, including the ponticulus. Moreover, GABA was shown to stimulate in vitro mango pollen tube elongation. The results support the heterotrophic growth of mango pollen tubes in the style at the expense of starch, similarly to the observations in porogamous species. However, unlike porogamous species, the micropyle of mango does not show high levels of GABA and starch, although they were observed in the ponticulus and could play a role in supporting the unusual pollen tube growth in chalazogamous species.

Abortion Categories and Characteristics of Acarpous Crape Myrtle Floral Organs

Lagerstroemia indica (crape myrtle) is a popular Chinese landscape plant with a long flowering period that contributes to its gorgeous flowers and high ornamental value, which motivate L. indica breeding. We found a wild acarpous individual of L. indica that did not bear seeds after flowering and had a significantly longer flowering period than fructiferous L. indica. This study identified differences in floral organ morphology, and stamen and pistil structure between fructiferous and acarpous L. indica through observation, paraffin sectioning, and scanning electron microscopy (SEM). The flowering time of each acarpous L. indica inflorescence lasts as long as 18 to 25 days. When a single flower withers, it falls from the pedicel without any fruit. The abortion in the floral organ of acarpous L. indica is characterized by sterile and undehisced anthers, pollen abortion, and deformed and irregularly arranged filament cells. Acarpous L. indica features short and loosely arranged papilla cells in the stigma, a flat style and narrow stylar canal, loosely arranged epidermal cells, and no obvious nuclei. No embryo sac cavity is found in acarpous L. indica ovules. In some nucelli, the egg apparatus structure can be observed indistinctly but without cell contour. In others, the egg apparatus structure is completely absent, and only flocculent tissue is observed. This study may provide a theoretical foundation for future studies on the molecular mechanisms of the mutations in acarpous L. indica.

Characteristics of Developmental Differences between Fertile and Aborted Ovules in Camellia oleifera

Camellia oleifera is an important woody edible oil plant in southern China. In this study, the developmental differences in ovules at different positions in the ovary of C. oleifera were observed. The developmental type and characteristics of aborted ovules, ratios of normal and aborted ovules, and their developmental differences after flowering were examined. Ovules near the stylar end and in the middle exhibit normal development and are able to form embryo sacs; lower ovules near the pedicel end are usually aborted. The proportion of abortion of four examined cultivars ranges from 10.2% to 33.3%. Aborted ovules can be divided into four categories: 1) nascent egg apparatus lacking distinguishable cells; 2) completely absent egg apparatus structure consisting of flocculent tissue; 3) lack of tissue, comprising only integument cells; and 4) the inner integument not constituting a micropyle channel, with incomplete egg apparatus development and generating abnormal ovules. At 120 days after pollination (DAP), significant distinguishable size differences were found between fertile and aborted ovules; aborted ovules ceased growth at 180 DAP. On fruit maturation, aborted seeds were still attached to the placenta.

Generative reproduction of Antarctic grasses, the native species Deschampsia antarctica Desv. and the alien species Poa annua L.

The embryology of two species, Deschampsia antarctica, a native species, and Poa annua, an alien species in the Antarctic we studied. Flowering buds of plants growing in their natural habitats on King George Island and generative tissues of both plant species grown in a greenhouse were analyzed. Adaptations to autogamy and anemogamy were observed in the flower anatomy of both species. The microsporangia of the evaluated grasses produce a small number of three−celled pollen grains. Numerous pollen grains do not leave the microsporangium and germinate in the thecae. Deschampsia antarctica and P. annua plants harvested in Antarctica developed a particularly small number of microspores in pollen chambers. In D. antarctica, male gametophytes were produced at a faster rate: generative cells in pollen did not become detached from the wall of the pollen grain, they were not embedded in the cytoplasm of vegetative cells, and they divided into two sperm cells situated close to the wall. The monosporous Polygonum type of embryo sac development was observed in the studied species. The egg apparatus had typical polarization, and the filiform apparatus did not develop in synergids. Large antipodals with polyploidal nuclei were formed in the embryo sacs of D. antarctica and P. annua. Poa annua was characterized by numerous antipodal cells which formed antipodal tissue in the chalazal region of the embryo sac. Three distinct antipodals with atypical, lateral position in the vicinity of the egg apparatus were observed in D. antarctica. The diaspores of the investigated grass species were characterized by small size, low weight and species-specific primary and secondary sculpture of the testa and caryopsis coat.

Ultrastructure of the egg apparatus of Spinacia

The egg apparatus of <em>Spinacia</em> was studied from the time the embryo sac reaches its maximal size to just before fertilization, i.e., until about 8-9 hours after pollination. At maturity each synergid has a large elongated nucleus and prominent chalazal vacuoles, Numerous mitochondria, plastids, dictyosomes, free ribosomes, rough endoplasmic reticulum (RER), and lipid bodies are present. The cell wall exists only around the micropylar half of the synergids and each cell has a distinct, striated filiform apparatus. In general, degeneration of one synergid starts after pollination. The egg cell has a spherical nucleus and nucleolus and a large micropylar vacuole. Numerous mitochondria, some plastids with starch grains, dictyosomes, free ribosomes, and HER are present. A continuous cell wall is absent around the chalazal end of the egg cell.

Polyembryony in maize: histological analysis

The histological analysis enabled to distinguish the main types of polyembryony in maize according to the origin of embryos connected with their localization in caryopses, differences in their structure (presence of common tissues) and type of germination. Twin embryos from multiple embryo sacs are usually localized on opposite sides (or distant places) of the caryopsis, they have no common tissues and the germination is separate. Twins or triplets from individual cells of the egg apparatus or multiple egg cells are closely adherent, but strictly separated by epidermal layers. Endosperm is common. Plumules and radicles are independent. Cleavage polyembryos arising <em>in vivo</em> spontaneously or after induction share a common suspensor, part of scutellum and surface layers of radicles. Therefore they germinate with one radicular complex and with separated plumules.