sidecar pollen, an Arabidopsis thaliana male gametophytic mutant with aberrant cell divisions during pollen development

Development ◽  
1996 ◽  
Vol 122 (10) ◽  
pp. 3243-3253 ◽  
Author(s):  
Y.C. Chen ◽  
S. McCormick

During pollen development each product of meiosis undergoes a stereotypical pattern of cell divisions to give rise to a three-celled gametophyte, the pollen grain. First an asymmetric mitosis generates a larger vegetative cell and a smaller generative cell, then the generative cell undergoes a second mitosis to give rise to two sperm cells. It is unknown how this pattern of cell divisions is controlled. We have identified an Arabidopsis gene, SIDECAR POLLEN, which is required for the normal cell division pattern during pollen development. In the genetic background of the NoO ecotype, sidecar pollen heterozygotes have about 45% wild-type pollen, 48% aborted pollen and 7% pollen with an extra cell. Homozygous sidecar pollen plants have about 20% wild-type pollen, 53% aborted pollen and 27% extra-celled pollen. Similar ratios of sidecar pollen phenotypes are seen in the Columbia ecotype but sidecar pollen is a gametophytic lethal in the Landsberg erecta ecotype. Thus this allele of sidecar pollen shows differential gametophytic penetrance and variable expressivity in different genetic backgrounds. The extra cell has the cell identity of a vegetative cell and is produced prior to any asymmetric microspore mitosis. Pollen tetrad analysis directly demonstrates that SIDECAR POLLEN is indeed expressed in male gametophytes. To our knowledge, scp is the first male gametophytic mutation to be described in Arabidopsis.

1992 ◽  
Vol 70 (7) ◽  
pp. 1369-1376 ◽  
Author(s):  
G. Hause ◽  
B. Hause ◽  
A. A. M. Van Lammeren

The structures of the microtubular and microfilamental cytoskeletons were investigated during the development of microspores and pollen grains of Brassica napus L. cv. Topas. Microfilaments were observed directly with rhodamine–phalloidin and microtubules with FITC by indirect immunofluorescent staining and transmission electron microscopy. We observed microtubules in all developmental stages and noted several changes in the configuration of the microtubular cytoskeleton during microspore development, microspore mitosis, and pollen development. A preprophase band before microspore mitosis was not observed. The arrest of the microspore nucleus in an eccentric position is likely caused by microtubules as is the shape of the phragmoplast at microspore mitosis. Despite the application of various staining methods, i.e., labelling of fixed and unfixed fresh and cryosectioned microspores and pollen with rhodamine–phalloidin, microfilaments could not be observed in all developmental stages. Prominent microfilamental arrays were observed during cytokinesis of microspore mitosis and during the free generative cell stage. They mark the stages with different configurations. Key words: Brassica napus, immunolabelling, cytoskeleton, microspore and pollen development.


Author(s):  
John R. Rowley

The morphology of the exine of many pollen grains, at the time of flowering, is such that one can suppose that transport of substances through the exine occurred during pollen development. Holes or channels, microscopic to submicroscopic, are described for a large number of grains. An inner part of the exine of Epilobium angustifolium L. and E. montanum L., which may be referred to as the endexine, has irregularly shaped channels early in pollen development although by microspore mitosis there is no indication of such channeling in chemically fixed material. The nucleus in microspores used in the experiment reported here was in prophase of microspore mitosis and the endexine, while lamellated in untreated grains, did not contain irregularly shaped channels. Untreated material from the same part of the inflorescence as iron treated stamens was examined following fixation with 0.1M glutaraldehyde in cacodylate-HCl buffer at pH 6.9 (315 milliosmoles) for 24 hrs, 4% formaldehyde in phosphate buffer at pH 7.2 (1,300 milliosmoles) for 12 hrs, 1% glutaraldehyde mixed with 0.1% osmium tetroxide for 20 min, osmium tetroxide in deionized water for 2 hrs and 1% glutaraldehyde mixed with 4% formaldehyde in 0.1M cacodylate-HCl buffer at pH 6.9 for two hrs.


2021 ◽  
Vol 7 (4) ◽  
pp. eabe2299 ◽  
Author(s):  
Na Wang ◽  
Jonathan I. Gent ◽  
R. Kelly Dawe

The production of haploids is an important first step in creating many new plant varieties. One approach used in Arabidopsis involves crossing plants expressing different forms of centromeric histone H3 (CENP-A/CENH3) and subsequent loss of genome with weaker centromeres. However, the method has been ineffective in crop plants. Here, we describe a greatly simplified method based on crossing maize lines that are heterozygous for a cenh3 null mutation. Crossing +/cenh3 to wild-type plants in both directions yielded haploid progeny. Genome elimination was determined by the cenh3 genotype of the gametophyte, suggesting that centromere failure is caused by CENH3 dilution during the postmeiotic cell divisions that precede gamete formation. The cenh3 haploid inducer works as a vigorous hybrid and can be transferred to other lines in a single cross, making it versatile for a variety of applications.


Genetics ◽  
2002 ◽  
Vol 160 (1) ◽  
pp. 63-73
Author(s):  
Michael J McEachern ◽  
Dana Hager Underwood ◽  
Elizabeth H Blackburn

Abstract Telomerase adds telomeric DNA repeats to telomeric termini using a sequence within its RNA subunit as a template. We characterized two mutations in the Kluyveromyces lactis telomerase RNA gene (TER1) template. Each initially produced normally regulated telomeres. One mutation, ter1-AA, had a cryptic defect in length regulation that was apparent only if the mutant gene was transformed into a TER1 deletion strain to permit extensive replacement of basal wild-type repeats with mutant repeats. This mutant differs from previously studied delayed elongation mutants in a number of properties. The second mutation, TER1-Bcl, which generates a BclI restriction site in newly synthesized telomeric repeats, was indistinguishable from wild type in all phenotypes assayed: cell growth, telomere length, and in vivo telomerase fidelity. TER1-Bcl cells demonstrated that the outer halves of the telomeric repeat tracts turn over within a few hundred cell divisions, while the innermost few repeats typically resisted turnover for at least 3000 cell divisions. Similarly deep but incomplete turnover was also observed in two other TER1 template mutants with highly elongated telomeres. These results indicate that most DNA turnover in functionally normal telomeres is due to gradual replicative sequence loss and additions by telomerase but that there are other processes that also contribute to turnover.


Development ◽  
1997 ◽  
Vol 124 (15) ◽  
pp. 2875-2888 ◽  
Author(s):  
L.A. Wrischnik ◽  
C.J. Kenyon

In C. elegans, six lateral epidermal stem cells, the seam cells V1-V6, are located in a row along the anterior-posterior (A/P) body axis. Anterior seam cells (V1-V4) undergo a fairly simple sequence of stem cell divisions and generate only epidermal cells. Posterior seam cells (V5 and V6) undergo a more complicated sequence of cell divisions that include additional rounds of stem cell proliferation and the production of neural as well as epidermal cells. In the wild type, activity of the gene lin-22 allows V1-V4 to generate their normal epidermal lineages rather than V5-like lineages. lin-22 activity is also required to prevent additional neurons from being produced by one branch of the V5 lineage. We find that the lin-22 gene exhibits homology to the Drosophila gene hairy, and that lin-22 activity represses neural development within the V5 lineage by blocking expression of the posterior-specific Hox gene mab-5 in specific cells. In addition, in order to prevent anterior V cells from generating V5-like lineages, wild-type lin-22 gene activity must inhibit (directly or indirectly) at least five downstream regulatory gene activities. In anterior body regions, lin-22(+) inhibits expression of the Hox gene mab-5. It also inhibits the activity of the achaete-scute homolog lin-32 and an unidentified gene that we postulate regulates stem cell division. Each of these three genes is required for the expression of a different piece of the ectopic V5-like lineages generated in lin-22 mutants. In addition, lin-22 activity prevents two other Hox genes, lin-39 and egl-5, from acquiring new activities within their normal domains of function along the A/P body axis. Some, but not all, of the patterning activities of lin-22 in C. elegans resemble those of hairy in Drosophila.


2014 ◽  
Vol 50 (3) ◽  
pp. 367-380 ◽  
Author(s):  
Elżbieta Bednarska

DNA and histone synthesis in five consecutive morphological stages of <em>Hyacinthus orientalis</em> L. pollen grain differentiation were studied autoradiographically. DNA synthesis was found to occur in both the generative and the vegetative cell. DNA replication in the generative cell took place when the generative cell was still adhered to the pollen grain wall but already devoid of callose wall. DNA synthesis in the generative cell slightly preceded that in the vegetative cell. Histones were synthesized in phase S of the generative and vegetative cell. In the generative cell histone synthesis also continued at a lower level after completion of DNA replication. In the developmental stages under study the nuclei of the generative cells were decidedly richer in lysine histones than vegetative cell nuclei.


2009 ◽  
Vol 2009 ◽  
pp. 1-12 ◽  
Author(s):  
K. Zienkiewicz ◽  
E. Bednarska

The aim of the present work was the characterization of nuclear bodies in the microspore and developing pollen cells ofHyacinthus orientalisL.. The combination of Ag-NOR, immunofluorescence and immunogold techniques was used in this study. The obtained results showed the presence of highly agyrophylic extranucleolar bodies in microspore and developing pollen cells, which were finally identified as Cajal bodies. In all cases, a strong accumulation of snRNP-indicating molecules including TMG cap, Sm proteins and U2 snRNA, was observed in the examined nuclear bodies. In contrast to their number the size of the identified structures did not change significantly during pollen development. In the microspore and the vegetative cell of pollen grains CBs were more numerous than in the generative cell. At later stages of pollen development, a drastic decrease in CB number was observed and, just before anthesis, a complete lack of these structures was indicated in both pollen nuclei. On the basis of these results, as well as our previous studies, we postulate a strong relationship between Cajal body numbers and the levels of RNA synthesis and splicing machinery elements in microspore and developing pollen cells.


Development ◽  
2001 ◽  
Vol 128 (4) ◽  
pp. 581-590 ◽  
Author(s):  
M. Herman

In Caenorhabditis elegans, Wnt signaling pathways are important in controlling cell polarity and cell migrations. In the embryo, a novel Wnt pathway functions through a (beta)-catenin homolog, WRM-1, to downregulate the levels of POP-1/Tcf in the posterior daughter of the EMS blastomere. The level of POP-1 is also lower in the posterior daughters of many anteroposterior asymmetric cell divisions during development. I have found that this is the case for of a pair of postembryonic blast cells in the tail. In wild-type animals, the level of POP-1 is lower in the posterior daughters of the two T cells, TL and TR. Furthermore, in lin-44/Wnt mutants, in which the polarities of the T cell divisions are frequently reversed, the level of POP-1 is frequently lower in the anterior daughters of the T cells. I have used a novel RNA-mediated interference technique to interfere specifically with pop-1 zygotic function and have determined that pop-1 is required for wild-type T cell polarity. Surprisingly, none of the three C. elegans (beta)-catenin homologs appeared to function with POP-1 to control T cell polarity. Wnt signaling by EGL-20/Wnt controls the migration of the descendants of the QL neuroblast by regulating the expression the Hox gene mab-5. Interfering with pop-1 zygotic function caused defects in the migration of the QL descendants that mimicked the defects in egl-20/Wnt mutants and blocked the expression of mab-5. This suggests that POP-1 functions in the canonical Wnt pathway to control QL descendant migration and in novel Wnt pathways to control EMS and T cell polarities.


Development ◽  
2000 ◽  
Vol 127 (18) ◽  
pp. 3931-3940 ◽  
Author(s):  
J.D. Walker ◽  
D.G. Oppenheimer ◽  
J. Concienne ◽  
J.C. Larkin

Cell differentiation is generally tightly coordinated with the cell cycle, typically resulting in a nondividing cell with a unique differentiated morphology. The unicellular trichomes of Arabidopsis are a well-established model for the study of plant cell differentiation. Here, we describe a new genetic locus, SIAMESE (SIM), required for coordinating cell division and cell differentiation during the development of Arabidopsis trichomes (epidermal hairs). A recessive mutation in the sim locus on chromosome 5 results in clusters of adjacent trichomes that appeared to be morphologically identical ‘twins’. Upon closer inspection, the sim mutant was found to produce multicellular trichomes in contrast to the unicellular trichomes produced by wild-type (WT) plants. Mutant trichomes consisting of up to 15 cells have been observed. Scanning electron microscopy of developing sim trichomes suggests that the cell divisions occur very early in the development of mutant trichomes. WT trichome nuclei continue to replicate their DNA after mitosis and cytokinesis have ceased, and as a consequence have a DNA content much greater than 2C. This phenomenon is known as endoreduplication. Individual nuclei of sim trichomes have a reduced level of endoreduplication relative to WT trichome nuclei. Endoreduplication is also reduced in dark-grown sim hypocotyls relative to WT, but not in light-grown hypocotyls. Double mutants of sim with either of two other mutants affecting endoreduplication, triptychon (try) and glabra3 (gl3) are consistent with a function for SIM in endoreduplication. SIM may function as a repressor of mitosis in the endoreduplication cell cycle. Additionally, the relatively normal morphology of multicellular sim trichomes indicates that trichome morphogenesis can occur relatively normally even when the trichome precursor cell continues to divide. The sim mutant phenotype also has implications for the evolution of multicellular trichomes.


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