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Changes in the microtubular cytoskeleton of Dactylorhiza majalis (Rchb.) Hunt et Summerh. (Orchidaceae) during microsporogenesis were investigated by the immunofluorescence method. Dactylorhiza microsporocytes in microsporangia divide into tetrads after a regular meiosis, which is completed with simultaneous cytokinesis. Three typical configurations of the microtubular cytoskeleton appear during microsporogenesis: the microtubular system in the cytoplasm and at the nuclear envelope, meiotic spindles, and phragmoplasts. Microtubules of the early prophase I microsporocyte are dispersed throughout the cortical cytoplasm, and later the arrays of microtubules are visible at the nuclear envelope. During metaphase I, the microtubules form the spindle which also acts during anaphase I. At telophase I, interzonal microtubules of the first meiotic spindle disappear, and new microtubular arrays extend from the nuclei towards the equatorial plane of the microsporocyte. There, these microtubules form the phragmoplast, which disintegrates before the second meiotic division. During the second meiotic division, the microtubular cytoskeleton repeats the configurations from the first meiotic division. The microtubular arrays emanating from the telophase II nuclei form interconnections of all non-sister and sister nuclei. During the formation of the cell plates between the future microspores, these microtubular arrays disappear. The results support the view that cytoskeletal configurations participate in the formation of the nuclear-cytoplasmic domains of the dividing microsporocyte in its transition from mononucleate microsporocyte to undivided dyad, and to four microspore domains after the second meiotic division.
Colobanthus quitensis (Kunth) Bartl, produced two types very small bisexual flowers. In the Antarctic natural conditions chasmogamic and cleistogamic flowers most often form five stamina with short filaments. Two microsporangia with a three-layer wall form in the anther. Microspore mother cells, which develop into microspores after meiosis, form inside the microsporangium. Microsporocytes of Colobanthus quitensis are surrounded with a thick callose layer, the special wall. After meiosis, the callose wall is dissolved and microspores are released from the tetrad. The production of proorbicules, orbicules and peritapetal membrane, and the construction of a complex sporoderm with numerous apertural sites were observed. When microspore and pollen protoplasts underwent necrosis, probably as a result of temperature and osmotic stress, sporoderm layers formed around microspores, and the cell tapetum did not disintegrate. However, woody wall layers did not accumulate in endothecium cells.
The microtubular cytoskeleton in dividing microsporocytes and developing pollen grains of Gagea lutea (L.) Ker.-Gaw. (Liliaceae) was investigated with a modified indirect immunofluorescence method. Meiotic and mitotic stages were identified by DAPI staining. The microtubular cytoskeleton was compared in plants originating from natural localities and others grown in the laboratory. In natural conditions, microsporocytes and pollen grains of wild early-spring Gagea lutea plants are subjected to abiotic factors including cold exposure and lack of water. The persistent influence of these factors can disturb microtubular cytoskeleton functioning. The following disturbances were observed in the course of microsporogenesis and pollen development: abnormal chromosome configurations in the metaphase of meiosis I; abnormally divided dyads with irregular, radial microtubule systems around the nuclei; the formation of differently sized microspores with irregular shapes, and irregular division; and the formation of pollen grains with vacuoles abnormal for their development stage. Similar kinds of disturbances were observed after 1.5 months of cold treatment (4°C) and drying in the laboratory. These abiotic factors simulated in laboratory conditions caused more disturbances in the course of microsporogenesis and produced more frequent defective pollen grains than in the sample that had experienced cold and drying in natural conditions.
This study investigated patterns of cytoskeletal organization during microsporogenesis in Chondrilla juncea L., an autonomous apomict with a triploid chromosome number (2n = 15). The distribution of microtubules and organelles is not typical. The microtubules do not form a normal phragmoplast and consequently the organelle equatorial plate observed in many taxons is not present. The organelles are dispersed randomly in both the central and peripheral parts of the cytoplasm.
In onion, seven stages of umbel development were distinguished with regard to the size of umbels, flower buds and developmental stages of male and female gametophytes. At the beginning of blooming, the stage at which buds are most often collected for induction of gynogenesis, flower bud length ranged from 2.0 to 5.0 mm, and the stages from archesporial cell to mature embryo sac were observed in the ovules. Microspores and bicellular pollen grains were observed in the anthers. Flower bud size was not an accurate indicator of pollen development. Buds of the same size but originating from umbels at different developmental stages were characterized by different stages of pollen development.
The role of histone H1 in the regulation of flower growth and development and in microsporogenesis was investigated in previously obtained transgenic tobacco plants expressing the antisense histone H1 cDNA. These plants exhibited a characteristic male-sterility phenotype caused by altered proportions of the native histone H1 in chromatin. The type and frequency of chromosomal aberrations during male meiosis as well as the distortions affecting tetrad formation in transgenic plants are reported.
Calmodulin distribution was studied in the maturing male fertile and male sterile anther of Allium cepa L., using commercial monoclonal anti-CaM antibodies. The main sites of its occurrence were the tapetum and the generative cell lines. During microsporogenesis, calmodulin localization in the anthers of both onion lines was similar. Anti-CaM was localized in dividing microsporocytes, and low levels of it were found in the tapetum cytoplasm. Significant differences in calmodulin level and distribution between male fertile and male sterile anthers were observed after completion of meiosis. In the tapetum of the male fertile anther, throughout its existence, calmodulin level was higher than in the tapetum of the male sterile anther. The prematurely degenerating male sterile tapetum, starting from the free-microspore stage, did not bind anti-CaM. Differences in calmodulin localization between the functional and nonfunctional tapetum were found already in the tetrad stage, that is, before the start of degeneration of microspores. In functional tapetum cells, anti-CaM was concentrated in the cytoplasm on the loculus side; in the nonfunctional tapetum of the male sterile anther, calmodulin was evenly distributed over the cells. In the microspores of the male sterile line, calmodulin was dispersed in the cytoplasm. During pollen maturation, calmodulin was observed to undergo polarization - anti-CaM was localized mainly in the region of one of the pollen poles, near the plasma membrane. In the male sterile anther, progressive degeneration of microspores was associated with decreasing calmodulin level in the cytoplasm; microspores in the abortion stage did not bind anti-CaM.
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