Insemination of the archegonium and fertilization in Taxus baccata L

1988 ◽  
Vol 89 (4) ◽  
pp. 551-560
Author(s):  
ROGER I. PENNELL ◽  
PETER R. BELL

A study of fertilization in Taxus baccata in the electron microscope has revealed novel features. Insemination of the archegonium is facilitated by local perforation of the wall of the young pollen tube. Digestion of the wall begins before the pollen tube pierces the megaspore membrane but is not completed until its tip makes contact with the neck cells of the archegonium. As soon as a pore is formed a single sperm nucleus and some cytoplasm of the male gametophyte enter the archegonium. Which of the paired sperm nuclei move from the pollen tube into the archegonium appears to be a matter of chance. Close apposition of sperm nucleus and egg nucleus is followed by the formation of numerous points of contact between the two. The membranes fuse at these points and pores are rapidly formed. The progressive enlargement of these pores ultimately eliminates any partitions and yields the zygotic nucleus. There is a possibility that, as in some other gymnosperms, the plastids and mitochondria of the zygote come in part from the male gametophyte, but whether from the remains of the spermatogenous cell cytoplasm or from the. pollen tube lumen is not clear.

The development of the male gametophyte of Taxus baccata has been studied over a period of 20 weeks, from germination of the microspore in February to spermatogenesis in July. A few days after germination the microspore nucleus divides and a transverse wall forms at the equator cutting off the small generative cell and a large tube cell. The latter immediately begins to expand to form the pollen tube. The first division thus establishes the polarity of the gametophyte and the generative cell is regarded as proximal. The transverse wall is ephemeral, and within six weeks it has disappeared. The nucleus of the generative cell divides while still at the proximal pole. The two daughter nuclei are unequal in size, but they remain associated and together move distally. The larger nucleus eventually becomes the nucleus of the spermatogenous cell, and the smaller the sterile nucleus. The spermatogenous cell acquires a distinctive cytoplasm and becomes surrounded by a wall which arises de novo . The nucleus of the spermatogenous cell enlarges, but always remains towards one side of the cell so that at mitosis the spindle is contained within one hemisphere. After division the wall of the spermatogenous cell is ruptured and the two sperms are released as naked nuclei of equal size. The cytoplasm of the spermatogenous cell degenerates as it enters the tube, but remains recognizable until fertilization.


2015 ◽  
Vol 42 (2) ◽  
pp. 323-343 ◽  
Author(s):  
J. Telżyńska ◽  
H. Telżyński

After controlled pollination of <i>Helianthus annuus</i> L. florets, the whole course of fertilization is described and documented on 24 microphotos. The timing of events is evaluated. The average cell cycle in the proembryo is 2 hours and the nuclear cycle in endosperm - 60 minutes.Plasmoptysis is suggested as the mechanism of pollen tube opening in the synergid. The structure of the thread-like sperm nucleus is interpreted as an end to end union of chromosomes, and the morphological changes of the sperm nuclei are explained as folding and coiling, based on a spiralization mechanism of chromosomes. Cytochemical observations indicating ribosome formation in the course of the nuclear cycles in the endosperm are described. The mechanisms accelerating nuclear cycles in the endosperm are discussed.


Author(s):  
A. Sosa ◽  
L. Calzada

The dependence of nuclear metabolism on the function of the nuclear membrane is not well understood. Whether or not the function of the nuclear membrane is partial or totally responsible of the repressed template activity of human sperm nucleus has not at present been elucidated. One of the membrane-bound enzymatic activities which is concerned with the mechanisms whereby substances are thought to cross cell membranes is adenosintriphosphatase (ATPase). This prompted its characterization and distribution by high resolution photogrammetry on isolated human sperm nuclei. Isolated human spermatozoa nuclei were obtained as previously described. ATPase activity was demonstrated by the method of Wachstein and Meisel modified by Marchesi and Palade. ATPase activity was identified as dense and irregularly distributed granules confined to the internal leaflet of the nuclear membrane. Within the nucleus the appearance of the reaction product occurs as homogenous and dense precipitates in the interchromatin space.


2021 ◽  
Vol 40 (2) ◽  
pp. 205-222
Author(s):  
Monica Scali ◽  
Alessandra Moscatelli ◽  
Luca Bini ◽  
Elisabetta Onelli ◽  
Rita Vignani ◽  
...  

AbstractPollen tube elongation is characterized by a highly-polarized tip growth process dependent on an efficient vesicular transport system and largely mobilized by actin cytoskeleton. Pollen tubes are an ideal model system to study exocytosis, endocytosis, membrane recycling, and signaling network coordinating cellular processes, structural organization and vesicular trafficking activities required for tip growth. Proteomic analysis was applied to identifyNicotiana tabacumDifferentially Abundant Proteins (DAPs) after in vitro pollen tube treatment with membrane trafficking inhibitors Brefeldin A, Ikarugamycin and Wortmannin. Among roughly 360 proteins separated in two-dimensional gel electrophoresis, a total of 40 spots visibly changing between treated and control samples were identified by MALDI-TOF MS and LC–ESI–MS/MS analysis. The identified proteins were classified according to biological processes, and most proteins were related to pollen tube energy metabolism, including ammino acid synthesis and lipid metabolism, structural features of pollen tube growth as well modification and actin cytoskeleton organization, stress response, and protein degradation. In-depth analysis of proteins corresponding to energy-related pathways revealed the male gametophyte to be a reliable model of energy reservoir and dynamics.


Development ◽  
1976 ◽  
Vol 36 (1) ◽  
pp. 67-72
Author(s):  
M. N. Skoblina

The behaviour of sperm nuclei was studied both in the cytoplasm of intact toad oocytes undergoing maturation and the cytoplasm of oocytes matured without germinal vesicles. The behaviour of the nuclei of pronase-treated sperm injected in the mature egg cytoplasm was shown to be exactly similar to that of the sperm nucleus after fertilization, i.e. they swelled, synthesized DNA, and divided. No changes in such sperm nuclei could be detected in the cytoplasm of the oocytes matured without germinal vesicles.


Horticulturae ◽  
2021 ◽  
Vol 7 (12) ◽  
pp. 529
Author(s):  
Emmy Dhooghe ◽  
Dirk Reheul ◽  
Marie-Christine Van Labeke

Hybridization in flowering plants depends, in the first place, on the delivery of pollen to a receptive stigma and the subsequent growth of pollen tubes through the style to the ovary, where the sperm nucleus of the pollen grain can ultimately fertilize the egg cell. However, reproductive failure is often observed in distant crosses and is caused by pre- and/or post-zygotic barriers. In this study, the reproductive pre-fertilization barriers of intertribal crosses between Anemone coronaria L. and Ranunculus asiaticus L., both belonging to the Ranunculaceae, were investigated. Despite the incongruity of intertribal crosses between A. coronaria and R. asiaticus having been of low intensity at the stigmatic level, interstylar obstructions of the pollen tube growth occurred, which confirmed the presence of pre-fertilization barriers. We show that these barriers could be partially bypassed by combining pollination with a stigma treatment. More specifically, a significantly higher ratio of the pollen tube length to the total style length and a better seed set were observed when the stigma was treated with the auxin 2,4-dichlorophenoxyacetic acid (2,4-D, 1 mg.mL−1) together with the cytokinin kinetin (KIN, 0.5 mg.mL−1) 24 h after pollination, irrespective of the cross direction. More specifically, the stigma treatments with any form of auxin (combined or not combined with cytokinin) resulted in a full seed set, assuming an apomictic fruit set, because no pollination was needed to obtain these seeds.


1982 ◽  
Vol 53 (1) ◽  
pp. 227-244
Author(s):  
T.C. Rodman ◽  
F.H. Pruslin ◽  
V.G. Allfrey

A standardized cytological preparation of mature mouse sperm has been devised to serve as an in vitro system for probing the intra-ooplasmic molecular events of transformation of the fertilizing sperm. Two parameters of the early phase of transformation in vivo are defined at the resolution of the light microscope: deletion of sperm-unique nuclear proteins, detectable by immunofluorescence, and retention of homogeneity of the residual DNA complex, with intact chromatin boundaries detectable by ethidium bromide staining. These studies show that both parameters are conserved when in vitro sperm preparations are treated with NaCl under reducing conditions. The deletion of 2 different classes of the unique basic proteins of mouse sperm nuclei is specified by the NaCl concentration: 0.7 M-NaCl displaces the non-protamine class but not the protamines, while 1 M-NaCl displaces both. On the other hand the effects of treatment with trypsin at various concentrations and intervals are less consistent with the in vivo parameters, indicating fragmentation and displacement, not only of the sperm-unique basic proteins, but also of structural proteins believed to maintain the fundamental cohesive organization of the DNA matrix. These observations suggest that mechanisms other than proteolysis, e.g. localized changes in ionic concentrations, may participate in the post-fertilization displacement of the sperm-unique nuclear proteins in vivo. This study also supports the validity of the in vitro simulation as a model with which to probe the progression of transformation of the sperm nucleus to the zygote pronucleus.


1939 ◽  
Vol 16 (4) ◽  
pp. 409-424 ◽  
Author(s):  
EDWARD L. CHAMBERS

The observations described in this paper indicate that the sperm nucleus is carried to the centre of the egg by the progressive enlargement of a gelated body, the sperm aster, and that the egg nucleus is carried to the sperm nucleus by a centripetal cytoplasmic flow directed toward the astral lake, within which the sperm nucleus lies. The curved path which the egg nucleus describes results from the continued change in direction of a cytoplasmic flow streaming toward the moving astral lake. The egg nucleus, when within the aster, is not only carried towards the lake by cytoplasmic streaming, but it is also moved centrally by the migration of the sperm aster toward the approximate centre of the egg.


1986 ◽  
Vol 61 (6) ◽  
pp. 382-383 ◽  
Author(s):  
Gabriella Bergamini Mulcahy ◽  
David L. Mulcahy
Keyword(s):  

1986 ◽  
Vol 102 (3) ◽  
pp. 1039-1046 ◽  
Author(s):  
H J Clarke ◽  
Y Masui

Zona-free oocytes of the mouse were inseminated at prometaphase I or metaphase I of meiotic maturation in vitro, and the behavior of the sperm nuclei within the oocyte cytoplasm was examined. If the oocytes were penetrated by up to three sperm, maturation continued during subsequent incubation and became arrested at metaphase II. Meanwhile, each sperm nucleus underwent the following changes. First, the chromatin became slightly dispersed. By 6 h after insemination, this dispersed chromatin had become coalesced into a small mass, from which short chromosomal arms later became projected. Between 12 and 18 h after insemination, each mass of chromatin became resolved into 20 discrete metaphase chromosomes. In contrast, if oocytes were penetrated by four to six sperm, oocyte meiosis was arrested at metaphase I, and each sperm nucleus was transformed into a small mass of chromatin rather than into metaphase chromosomes. If oocytes were penetrated by more than six sperm, the maternal chromosomes became either decondensed or pycnotic, and the sperm nuclei were transformed into larger masses of chromatin. As control experiments, immature and fully mature metaphase II oocytes were inseminated. In the immature oocytes, which were kept immature by exposure to dibutyryl cyclic AMP, no morphological changes in the sperm nucleus were observed. On the other hand, in the fully mature oocytes, which were activated by sperm penetration, the sperm nucleus was transformed into the male pronucleus. Therefore, the cytoplasm of the maturing oocyte develops an activity that can transform the highly condensed chromatin of the sperm into metaphase chromosomes. However, the capacity of an oocyte is limited, such that it can transform a maximum of three sperm nuclei into metaphase chromosomes. Furthermore, the presence of more than six sperm causes a loss of the ability of the oocyte to maintain the maternal chromosomes in a metaphase state.


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