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12 recordsLinked to original sources
PubMed1991

Nucleic acid and protein synthesis during lateral root initiation in Marsilea quadrifolia (Marsileaceae).

The pattern of DNA, RNA, and protein synthesis during lateral root initiation in Marsilea quadrifolia L. was monitored by autoradiography of incorporated of 3H-thymidine, 3H-uridine, and 3H-leucine, respectively. DNA synthesis was associated with the enlargement of the lateral root initial prior to its division. Consistent with histological studies, derivatives of the lateral root initial as well as the cells of the adjacent inner cortex and pericycle of the parent root also continued to synthesize DNA. RNA and protein synthetic activities were found to be higher in the lateral root initials than in the endodermal initials of the same longitudinal layer. The data suggest a role for nucleic acid and protein synthesis during cytodifferentiation of a potential endodermal cell into a lateral root initial.

Autoradiography↗
PubMed1999

Phylogeny of Marsileaceous Ferns and Relationships of the Fossil Hydropteris pinnata Reconsidered.

Recent phylogenetic studies have provided compelling evidence that confirms the once disputed hypothesis of monophyly for heterosporous leptosporangiate ferns (Marsileaceae and Salviniaceae). Hypotheses for relationships among the three genera of Marsileaceae (Marsilea, Regnellidium, and Pilularia), however, have continued to be in conflict. The phylogeny of Marsileaceae is investigated here using information from morphology and rbcL sequence data. In addition, relationships among all heterosporous ferns, including the whole-plant fossil Hydropteris pinnata are reconsidered. Data sets of 71 morphological and 1239 rbcL characters for 23 leptosporangiate ferns, including eight heterosporous ingroup taxa and 15 homosporous outgroup taxa, were subjected to maximum parsimony analysis. Morphological analyses were carried out both with and without the fossil Hydropteris, and it was excluded from all analyses with rbcL data. An annotated list of the 71 morphological characters is provided in the appendix. For comparative purposes, the Rothwell and Stockey (1994) data set was also reanalyzed here. The best estimate of phylogenetic relationships for Marsileaceae in all analyses is that Pilularia and Regnellidium are sister taxa and Marsilea is sister to that clade. Morphological synapomorphies for various nodes are discussed. Analyses that included Hydropteris resulted in two most-parsimonious trees that differ only in the placement of the fossil. One topology is identical to the relationship found by Rothwell and Stockey (1994), placing the fossil sister to the Azolla plus Salvinia clade. The alternative topology places Hydropteris as the most basal member of the heterosporous fern clade. Equivocal interpretations for character evolution in heterosporous ferns are discussed in the context of these two most-parsimonious trees. Because of the observed degree of character ambiguity, the phylogenetic placement of Hydropteris is best viewed as unresolved, and recognition of the suborder Hydropteridineae, as circumscribed by Rothwell and Stockey (1994), is regarded as premature. The two competing hypotheses of relationships for heterosporous ferns are also compared with the known temporal distribution of relevant taxa. Stratigraphic fit of the phylogenetic estimates is measured by using the Stratigraphic Consistency Index and by comparison with minimum divergence times.

Journal Article↗
PubMed1979

Developmental mechanisms in heterospory: cytochemical demonstration of spore-wall enzymes associated with beta-lectins, polysaccharides and lipids in water ferns.

Cytochemical methods are used to examine the distribution and localization of acid phosphatase, non-specific esterase, ribonuclease and peroxidase activity in the walls of the spores of the heterosporous Marsileaceae before and during germination. In the quiescent spore, the principal activity is associated with the perine layer of the wall and the intine, with some activity in the outer, gelatinous wall layer, but none in the exine. The microspores of Marsilea and Pilularia have non-specific esterase activity concentrated in the intine inthe immediate vicinity of the germinal site; that is, above the position of the future male gametangia. The enzymes are not leached from the wall during hydration of the spores, although ribonuclease is redistributed during imbibition with a high concentration of activity remaining in or around the germinal site. The wall enzymes occur together with PAS-reactive and acidic carbohydrates, lipids, and in the microspore perine, beta-lectins. Following the enzyme pattern, the beta-lectins are found to be concentrated in the region of the germinal site. beta-Lectin activity is absent from the megaspore wall. Acidic carbohydrates are confined to the gelatinous wall layer and this layer also binds concanavalin A. In contrast to what has been found for other plant cells, the spore-wall beta-lectins are not water-labile; the activity is not significantly diminished after hydration. This surprising stability suggests that these molecules, together with the enzymes, may be retained in position in the wall by the waterproof overlay of lipid. From the evidence of preliminary developmental studies, it appears that the enzymes associated with the perine layer of the wall originate in the sporophytic tapetal periplasmodium and inclusion of the activity is concurrent with wall differentiation, while the activity associated with the intine is derived from the gametophyte. It is possible, however, in the megaspore at least, that the distribution of the activity may to some extent be influenced by a system of exine channels which communicates between the two domains of the wall during sporogenesis. No definite information is obtained concerning the utility of the enzymes and associated molecules in the life of the spore. Acting separately or in co-operation, their role could conceivably be connected with one or more of four processes; wall differentiation, gametophyte nutrition, gametophyte protection or reproduction. Each of these possibilities is discussed.

Acid Phosphatase↗
PubMed2003

Changes in the abundance and distribution of conserved centrosomal, cytoskeletal and ciliary proteins during spermiogenesis in Marsilea vestita.

Spermiogenesis in the male gametophytes of the water fern Marsilea vestita is a precise and rapid process resulting in the production of ciliated gametes. Development begins from a single cell within the microspore wall that undergoes nine rapid cell division cycles in distinct planes to produce 32 spermatids that are surrounded by 7 sterile cells. Thereafter, the de novo formation of basal bodies occurs in a discrete cytoplasmic particle known as a blepharoplast, with the subsequent formation of a complex ciliary apparatus in elongating spermatids. The rate and extent of development appear to be controlled at a post-transcriptional level, where the sudden translation of specific stored mRNAs (e.g., centrin) results in the formation of particular structures in the cells (e.g., blepharoplasts). We show here that additional centrosomal and cytoskeletal antigens known as SF assemblin, p95 kDa protein, delta tubulin, gamma tubulin, Xgrip109, Aik, CTR453, RanBPM, BX63, RSP6, and alpha tubulin each exhibit specific localization patterns both on immunoblots of gametophyte protein isolates and in fixed cells. BAp90, PP4, and RLC exhibit specific localization patterns in fixed cells. We show that the antigens exhibit complex patterns of abundance during spermiogenesis. In an attempt to identify regulatory agents involved in spermiogenesis, we employed a RNAi-based screen of 41 randomly selected gametophyte cDNAs on developing populations of synchronously growing gametophytes. The gametophytes treated with each of the RNAi probes exhibited arrest at a specific stage of development. A consequence of anomalous development was the block to assembly of the ciliary apparatus, an effect highlighted by altered staining with anti-centrin, anti-beta-tubulin, and anti-RSP6 antibodies. Our results show that complex, integrated processes of translation and protein partitioning apparently underlie the assembly of the ciliary apparatus during spermiogenesis in male gametophytes of M. vestita.

Antibodies↗
PubMed2004

Differential segregation and modification of mRNA during spermiogenesis in Marsilea vestita.

We are interested in the mechanisms that underlie cell fate determination in the endosporic male gametophytes of the fern, Marsilea vestita. Synchronous development is initiated by placing dry spores into water and involves the translation of stored mRNAs, with little transcription. Nine division cycles produce 32 spermatids surrounded by 7 sterile cells, and then each spermatid differentiates into a multiciliate gamete. Here, we focus on changes in the distribution of particular proteins, mRNAs, and patterns of polyadenylation as essential prerequisites for cell fate determination and gametogenesis. Earlier, we showed that alpha- and beta-tubulin proteins become concentrated in spermatogenous initials, and that centrin mRNA is translated only in spermatogenous initials. In situ hybridizations reveal that centrin, cyclin B, and beta-tubulin mRNAs are present in both sterile and spermatogenous cells, but that transcripts encoding RNA helicase and PRP-19 (a spliceosome component) become localized in spermatogenous cells. The targeted destruction of these two transcripts by RNAi treatments does not affect the numbers of division cycles, but the gametophytes exhibit anomalous patterns of cytokinesis, and a subsequent failure of spermatid differentiation. Thus, cell fate determination in the gametophyte involves localized translation, and the localization of mRNAs for proteins involved in transcript processing. We found differences in polyadenylation levels in sterile and spermatogenous cells that match the distribution of cytoplasmic poly(A) polymerase (PAP), which, in immunolocalizations, is abundant in spermatogenous cells, but undetectable in sterile cells. The activation of translation in spermatogenous initials, but not in sterile cells, may be under the control of mRNA processing enzymes, which become localized either as proteins or mRNAs in the spermatogenous subdomains before any divisions occur.

Cell Differentiation↗
PubMed2005

Abscisic acid regulation of heterophylly in Marsilea quadrifolia L.: effects of R-(-) and S-(+) isomers.

The plant hormone abscisic acid (ABA) induces a developmental switch in the aquatic fern Marsilea quadrifolia, causing the formation of aerial type characteristics, including the elongation of petioles and roots, a change in leaf morphology, the expansion of leaf surface area, and the shortening of the internodes. A number of ABA-responsive heterophylly (ABRH) genes are induced early during the transition. Using optically pure isomers of ABA, it was found that both the natural S-(+)-ABA and the unnatural R-(-)-ABA are capable of inducing a heterophyllous switch and regulating ABRH gene expression. When dose responses are compared, the unnatural ABA gives stronger morphogenic effects than the natural ABA at the same concentration, it is effective at lower concentrations, and its optimal concentration is also lower compared with the natural ABA. Deuterium-labelled ABA enantiomers were used to trace the fate of applied ABA and to distinguish the applied compound and its metabolites from the endogenous counterparts. In tissues, the supplied (+)-ABA was metabolized principally to dihydrophaseic acid, while the supplied (-)-ABA was converted at a slower rate to 7'-hydroxy abscisic acid. Treatment with either enantiomer resulted in increased biosynthesis of ABA, as reflected in the accumulation of endogenous dihydrophaseic acid. Taken together, these results suggest two distinct mechanisms of action for (-)-ABA: either (-)-ABA is intrinsically active, or its activity is due to the stimulation of ABA biosynthesis.

Abscisic Acid↗
PubMed2004

An easy and versatile embedding method for transverse sections.

In several research areas, transverse sections are indispensable for studying structural aspects of specimens. However, the oriented embedding of small cylindrical samples can become problematic, especially when transverse sections at right angles to the main axis of the object are desired. Here, we describe an easy and low-cost technique for oriented embedding of small (psi < 500 micro m) as well as of larger specimens (psi > 500 micro m). The usefulness of the technique is demonstrated for roots and stamens of Arabidopsis thaliana and for adventitious roots of Asplenium demerkense, as examples of small and larger cylindrical samples, respectively. Furthermore, several types of resin (glycol methacrylate, epoxy and acrylic resins) were successfully tested, showing the applicability of the technique for light and electron microscopy and for immunolocalizations. In conclusion, the principle of the technique can be extended to several resins and a wide variety of specimen types, such as stems, leaves and textile fibres. The originality of the technique lies in its simplicity combined with its high efficiency to produce well-oriented transverse sections.

Arabidopsis↗