TL;DR: Results suggest that gcH2A and gc H3 mRNAs are products of transcriptional activation of the generative cell nucleus and thus provide the first evidence for male germ line cell-specific gene expression in flowering plants.
Abstract: Formation of the generative cell in flowering plants initiates the male spermatogenesis pathway which eventually culminates in the process of double fertilization Little is known about the molecular mechanisms operative in the generative cell Here, we report the isolation and characterization of cDNA clones encoding generative cell-specific histones, gcH2A and gcH3 In situ hybridization analysis revealed that both genes are activated after formation of the generative cell and accumulation of transcripts increases progressively during generative cell maturation These results suggest that gcH2A and gcH3 mRNAs are products of transcriptional activation of the generative cell nucleus and thus provide the first evidence for male germ line cell-specific gene expression in flowering plants
TL;DR: There is no requirement for sustained RNA synthesis by the generative cell nucleus for normal gametophytic development in pollen embryogenesis in cultured anthers of H. niger, it is concluded.
Abstract: The pattern of RNA synthesis during maturation and germination of pollen grains ofHyoscyamus niger was studied using3H-uridine autoradiography. Incorporation of label during pollen maturation was periodic with peak RNA synthesis occurring in the uninucleate, nonvacuolate pollen grains and in the vegetative cell of the bicellular pollen grains. During the early stages of germination, isotope incorporation occurred predominantly in the nucleus of the vegetative cell with little or no incorporation in the generative cell. With the appearance of the pollen tube, incorporation of3H-uridine in the vegetative cell nucleus decreased and completely disappeared at later stages of germination. No incorporation of isotope was observed in the sperms formed in the pollen tube by the division of the generative cell. From a comparison of the results of this study with those of previous works on RNA synthesis during pollen embryogenesis in cultured anthers ofH. niger, it is concluded that in contrast to embryogenic development, there is no requirement for sustained RNA synthesis by the generative cell nucleus for normal gametophytic development.
TL;DR: This study demonstrates that specific histone variants are present in the male gametic nucleus of a higher plant, as they are in the sperm nucleus of animals, and distinct differences in histone composition exist between the nuclei of generative and vegetative cells in pollen.
Abstract: Two proteins that resemble core histones and might be specific to the male gametic (generative) nucleus within the pollen of Lilium longiflorum Thumb, (originally designated p22.5 and p18.5; K. Ueda and I. Tanaka, 1994, Planta, 192, 446–452) were characterized biochemically and immunochemically. Patterns of digestion of p22.5 and p18.5 by Staphylococcus aureus V8 protease closely resembled those of somatic histones H2B and H3, respectively. However, peptide fragments that were unique to p22.5 or p18.5 were also detected. Antibodies raised against these proteins did not cross-react with any somatic histones. These results indicate that p22.5 and p18.5 are different from somatic histones in terms of primary structure. Analysis of their amino-acid compositions revealed that p22.5 is a moderately lysine-rich protein while p18.5 is an arginine-rich protein. From these results, we conclude that p22.5 is a variant of histone H2B and p18.5 is a variant of histone H3. Immunofluorescence staining of pollen grains using the specific antibodies revealed that both p22.5 and p18.5 are only present in the generative cell nucleus and are not to be found in the vegetative cell nucleus. This study demonstrates that (i) specific histone variants are present in the male gametic nucleus of a higher plant, as they are in the sperm nucleus of animals, and (ii) distinct differences in histone composition exist between the nuclei of generative and vegetative cells in pollen. These novel histones (p22.5 and p18.5), specific to male gametic nuclei, have been designated gH2B and gH3, respectively.
TL;DR: DNA synthetic patterns in the generative and vegetative cell nuclei confirm the ontogeny of embryoids described in this plant.
Abstract: Continued DNA synthesis in the generative cell nucleus, followed by mitosis and cytokinesis, results in the formation of pollen embryoids in cultured anthers of H. niger. In contrast, the nucleus of the vegetative cell undergoes no DNA synthesis after it is cut off, or synthesizes DNA only during a limited number of cell cycles. DNA synthetic patterns in the generative and vegetative cell nuclei confirm the ontogeny of embryoids described in this plant.
TL;DR: The reproduction system of this species represents a new type of single fertilization (non-double fertilization) between egg cell and sperm cell nuclei caused by the omission of generative cell division and the formation of only one sperm cell nucleus.
Abstract: The reproduction system inSpiranthes sinensis (Orchidaceae), collected at various localities in Japan, was revealed to be of a peculiar new type which is not to be found in other angiosperms. None of generative cell nuclei has been observed to dividede novo throughout the progamic phase, although they do participate in fertilization. Neither fertilization of the generative cell nucleus with the central nucleus nor formation of endosperm occurred in this plant. Although the chromosome number in the developing proembryonic cells numbered 2n=30, exhibiting diploidy, the number of marker chromosomes was equal to the sum of half the number of marker chromosomes of the parental clones.