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The meiotic division of microsporocytes and pollen grain development in Gagea lutea (L.) Ker.-Gaw. (Liliaceae) with fluorescence microscope (excitation light 400 nm) was observed after squashing the anthers in DAPI solution (fluorochrom dying DNA). Up to 70% of microsporocytes and pollen grains during the microsporogenesis and pollen grain development take a regular course. In the remaining microsporocytes and pollen grains (30%) the disturbances in course of both processes were observed. The most often observed disturbances are "late" chromosomes and a presence of micronuclei. The divisions of microsporocytes in the anther loculi show a big asynchrony, which, like the disturbances during the course of microsporogenesis and pollen grain development, may be caused by the external factors. The microsporogenesis takes place during autumn and winter months: the pollen grains develop in winter. At this time the dividing microsporocytes and developing pollen grains are under the influence of abiotic factors as low temperature and a lack of water. These factors disturb the formation of microtubular cytoskeleton of the dividing microsporocytes and pollen grains, which causes the formation of sterile pollen grains.
Immunogold labelling revealed the presence of lipoxygenase (LOX) in different parts and types of anther cells of Gagea lutea. LOX was found in the cytoplasm and close to ER elements in epidermal and endothecial cells, and close to the cell walls of the latter. The positive immunoreaction to LOX was less intense in the middle layers and the loculus of the anther, where single immunogold particles were concentrated at the cell walls of these layers and in the protoplast masses, in vacuoles, close to mitochondria, inside plastids, and in the liquid of the anther cavity. LOX occurred in the cytoplasm and around ER elements of pollen grains as well as in the exine layer, particularly in contact regions between the outer and inner exine layers. The correlations between LOX localization in different anther cells and the functioning of particular anther parts are discussed.
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.
Localization of lipoxygenase (LOX) in the microspore of Gagea lutea (L.) Ker.-Gaw. was investigated with the immunogold labelling method. The enzyme was found in the cytoplasm, nucleus and sporoderm. The most intensive reaction was observed in the cytoplasm, where the immunogold particles were sometimes grouped into clusters of several or more and showed the highest density. The smallest amount of particles occured in the sporoderm. The role of lipoxygenase in the microspore is discussed.
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