The mature embryo sac of wheat contains an egg apparatus composed of an egg cell and two synergids at the micropylar end, a central cell with two large polar nuclei in the middle, and a mass of 20 to 30 antipodals at the chalazal end. A comparison was made of the ultrastructural features of the various cells of the embryo sac. The features included the position of the nucleus and vacuoles, the number, structure, and distribution of organelles, and the extent of the cell walls surrounding each cell. The pollen tube enters one synergid through the filiform apparatus from the micropyle. The penetration and discharge of the pollen tube causes the further degeneration of that synergid, which had already undergone changes before pollination. The second synergid does not change further in appearance following the penetration of the first by the pollen-altered tube. Half an hour after pollination at 20–25 °C, two male nuclei are seen in the cytoplasm of the egg and the central cell. At about 1 h after pollination, one sperm has made contact with the egg nucleus, while the other sperm is fusing with one of the polar nuclei.
Many late embryogenesis abundant (Lea) protein genes in plants are regulated by abscisic acid (ABA). The RNA level of a carrot gene, DC8, increases in response to ABA in developing seeds. However, DC8 cannot be induced by ABA in adult tissues. We used chimeric genes made of various DC8 promoter fragments fused to beta-glucuronidase (GUS) to analyze the transcriptional regulation of DC8. DC8:GUS expression was measured in electroporated carrot protoplasts and in stably transformed carrots. The region of the DC8 promoter from -170 to -51 contained ABA-responsive sequences that required a 5' upstream region for high levels of expression in embryogenic callus protoplasts. 505 bp of the DC8 promoter conferred GUS expression in stably transformed somatic and zygotic embryos. DC8:GUS was expressed only in tissues formed in the seed. This includes cells in the embryo, the endosperm and the germinating seedlings. Gel retardation and competition experiments were performed to analyze the embryo nuclear protein-DNA binding activities in vitro. No binding activity was detected on the putative ABA-responsive region; however the 5' upstream regions located between -505 and -301 interacted with embryo nuclear factors. An additional site of DNA-protein interaction was located between positions -32 and +178. The nuclear proteins that bind these sequences were found in the embryo nuclei only, not in the nuclei from leaves or roots.
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