Studies on Vicia faba root meristems irradiated with a pion beam.
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Results obtained by use of cytophotometric, autoradiographic and electron-microscopic methods indicate that periodic changes occur during the course of the cell cycle in synthesis, accumulation and transport of rRNA. At the final periods of G1, S, and G2 phases an increased transport of rRNA into cytoplasm takes place; owing to an abrupt loss of nucleolar contents a transparent zone ("vacuole") appears and the majority of nucleoli becomes visible as ring-shaped. Due to an intensive synthesis and accumulation of rRNA some ring nucleoli transform into compact nucleoli during middle S and middle G2. Compact nucleoli which coexist with ring nucleoli also undergo periodic changes; they consist in the decrease in content and concentration of RNA by the final periods of G1, S, and prophase, as well as the reverse process, i.e. an increase in both content and concentration of RNA at early stages of S and G2.
The frequency of different types of pairs and triads of sister cells with labelled or unlabelled cells in mitosis was determined at different times after short-term incubation with 3H-thymidine to establish whether the mitotic cycle and G2-period duration depends on the cell length. Out of two sister cells of the pair, the larger cell used to have shorter mitotic cycle and G2-period than the smaller one. No direct dependence of the different ratios between their lengths was, however, shown. This is due to the absence of direct correlation between the mitotic cycle and G2-period durations. Possible causes of the absence of such a correlation are discussed.
Germination of seeds in cadmium chloride solution resulted in certain disturbances in nuclear division and stopped cytokinesis. Cystein added to cadmium chloride solution decreased the number of these disturbances. The effect of cadmium ions is supposed to be associated with the SH-groups blocking in contractile proteins of cellular spindle or in enzymes responsible for mitosis.
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Many higher plants have shoot apical meristems that possess discrete cell layers, only one of which normally gives rise to gametes following the transition from vegetative meristem to floral meristem. Consequently, when mutations occur in the meristems of sexually reproducing plants, they may or may not have an evolutionary impact, depending on the apical layer in which they reside. In order to determine whether developmentally sequestered mutations could be released by herbivory (i.e., meristem destruction), a characterized genetic mosaic was subjected to simulated herbivory. Many plants develop two shoot meristems in the leaf axils of some nodes, here referred to as the primary and secondary axillary meristems. Destruction of the terminal and primary axillary meristems led to the outgrowth of secondary axillary meristems. Seed derived from secondary axillary meristems was not always descended from the second apical cell layer of the terminal shoot meristem as is expected for terminal and primary shoot meristems. Vegetative and reproductive analysis indicated that secondary meristems did not maintain the same order of cell layers present in the terminal shoot meristem. In secondary meristems reproductively sequestered cell layers possessing mutant cells can be repositioned into gamete-forming cell layers, thereby adding mutant genes into the gene pool. Herbivores feeding on shoot tips may influence plant evolution by causing the outgrowth of secondary axillary meristems.
Root meristem structure and root branching in three species of Cladopus were investigated from developmental and anatomical perspectives. Cladopus fukiensis has a compressed bell-shaped meristem at the apex of a compressed subcylindrical root, while C. javanicus and perhaps C. nymanii, with a ribbon-like root, have a half lozenge-shaped ( subset as seen from above) meristem composed of an apical meristem of cubic cells and a marginal meristem of rectangular cells. The dorsiventrality of the meristem results in root dorsiventrality, and a marginal meristem contributes to the broadening of the root. Comparisons of meristem structure and root morphology suggest that the ribbon-like root of, e.g. C. javanicus, evolved towards the foliose root of Hydrobryum, sister to the genus Cladopus, by loss of an indeterminate apical meristem. The lateral root of C. javanicus initiates within the meristem of a parent root. The dorsal dermal layer and inner cells of the lateral-root meristem appear endogenously under the dermal layer of the parent root, while the ventral layer is derived exogenously from a ventral dermal layer continuous with the parent-root meristem. This mosaic pattern of exogenous and endogenous root formation differs from the truly exogenous formation seen in Hydrobryum and Zeylanidium. The dorsiventral mosaic origin of the root meristem may account for root cap asymmetry.
The flowering process in grapevine (Vitis vinifera) takes place in buds and extends for two consecutive growing seasons. To understand the genetic and molecular mechanisms underlying this process, we have characterized grapevine bud development, cloned the grapevine FLORICAULA/LEAFY (FLO/LFY) ortholog, VFL, and analyzed its expression patterns during vegetative and reproductive development. Flowering induction takes place during the first season. Upon induction, the shoot apical meristem begins to produce lateral meristems that will give rise to either inflorescences or tendrils. During the second season, after a winter dormancy period, buds reactivate and inflorescence meristems give rise to flower meristems. VFL is expressed in lateral meristems that give rise to inflorescence and flower meristems, consistent with a role in reproductive development. Furthermore, VFL is also detected in other meristematic regions such as the vegetative shoot apical meristem and the lateral meristems that will give rise to tendrils. VFL is also expressed in leaf primordia and in growing leaf margins until later stages of development. Accumulation of VFL transcripts in cell-proliferating regions suggests a role for VFL not only in flower meristem specification, but also in the maintenance of indeterminacy before the differentiation of derivatives of the apical meristem: flowers, leaves, or tendrils.
A main determinant of inflorescence architecture is the site where floral meristems are initiated. We show that in wild-type Petunia bifurcation of the inflorescence meristem yields two meristems of approximately equal size. One terminates into a floral meristem and the other maintains its inflorescence identity. By random transposon mutagenesis we have generated two mutants in which the architecture of the inflorescence is altered. In the extra petals- (exp) mutant the inflorescence terminates with the formation of a single terminal flower. Phenotypic analysis showed that exp is required for the bifurcation of inflorescence meristems. In contrast, the aberrant leaf and flower- (alf) mutant is affected in the specification of floral meristem identity while the branching pattern of the inflorescence remains unaltered. A weak alf allele was identified that, after bifurcation of the inflorescence meristem, yields a 'floral' meristem with partial inflorescence characteristics. By analysing independent transposon dTph1 insertion alleles we show that the alf locus encodes the Petunia FLORICAULA/LEAFY homolog. In situ hybridisation shows that alf is expressed in the floral meristem and also in the vegetative meristem. Differences and similarities between these Petunia mutants and mutations affecting inflorescence architecture in other species will be discussed.
In Arabidopsis, floral meristems arise in continuous succession directly on the flanks of the inflorescence meristem. Thus, the pathways that regulate inflorescence and floral meristem identity must operate both simultaneously and in close spatial proximity. The TERMINAL FLOWER 1 (TFL1) gene of Arabidopsis is required for normal inflorescence meristem function, and the LEAFY (LFY), APETALA 1 (AP1), and APETALA 2 (AP2) genes are required for normal floral meristem function. We present evidence that inflorescence meristem identity is promoted by TFL1 and that floral meristem identity is promoted by parallel developmental pathways, one defined by LFY and the other defined by AP1/AP2. Our analysis suggests that the acquisition of meristem identity during inflorescence development is mediated by antagonistic interactions between TFL1 and LFY and between TFL1 and AP1/AP2. Based on this study, we propose a simple model for the genetic regulation of inflorescence development in Arabidopsis. This model is discussed in relation to the proposed interactions between the inflorescence and the floral meristem identity genes and in regard to other genes that are likely to be part of the genetic hierarchy regulating the establishment and maintenance of inflorescence and floral meristems.
The narrow leaf and shortened stem phenotypes of the maize mutant narrow sheath (ns) are postulated to result from the lack of founder cell initialization in a region of the meristem that gives rise to leaf and stem margins. To test this model, a lineage map of the maize meristem is presented which compares the development of leaf margins in the narrow leaf mutant, narrow sheath (ns), and wild-type sibling plants. X-irradiation of mature seeds produced aneuploid albino sectors in wild-type and ns mutant plants. Of particular interest are sectors occurring in more than one leaf, which reflect a meristematic albino cell lineage. Analyses of these sectors indicated that: (1) a region of the ns meristem does not contribute to the founder cell population of the incipient leaf; (2) the margins of ns mutant leaves are derived from a lateral region of the meristem different from those in wild-type siblings; (3) founder cells in wild-type, juvenile-staged vegetative meristems encircle the meristem to a greater extent than do founder cells in adult-staged meristems; and (4) meristematic leaf founder cells may be subdivided into specific lateral domains, such that the position of a sector on the meristem correlates with a particular cell lineage. These data support our model for ns gene function in a specific domain of the meristem.
The mechanisms that establish the floral meristem are now becoming clearer, but the way in which flowering is maintained is less well understood. Impatiens balsamina provides a unique opportunity to address this question because reversion to vegetative growth can be obtained in a predictable way by transferring plants from inductive to non-inductive conditions. Following increasing amounts of induction, reversion takes place at progressively later stages of flower development. Partial flower induction and defoliation experiments show that a floral signal is produced in the cotyledon in response to inductive conditions and that this signal progressively diminishes after transfer to non-inductive conditions, during reversion. Therefore reversion in Impatiens is most likely due to the failure of leaves to become permanent sources of inductive signal in addition to the lack of meristem commitment to flowering. Analysis of the expression of the Impatiens homologues of the meristem identity genes floricaula and squamosa indicates that a change in floricaula transcription is not associated with the establishment or maintenance of the floral meristem in this species. Squamosa transcription is associated with floral development and petal initiation, and is maintained in existing petal or petaloid primordia even after the meristem has reverted. However, it is not expressed in the reverted meristem, in which leaves are initiated in whorled phyllotaxis and without axillary meristems, both characteristics usually associated with the floral meristem. These observations show that squamosa expression is not needed for the maintenance of these floral characters. The requirement for the production of the floral signal in the leaf during the process of flower development may reflect an additional function separate to that of squamosa activation; alternatively the signal may be required to ensure continued transcriptional activation in the meristem.
We have analyzed two mutants that exhibit altered panicle architecture in rice (Oryza sativa L.). In lax1-2, which is a new and stronger allele of the previously reported lax mutant, initiation and/or maintenance of rachis-branches, lateral spikelets, and terminal spikelets was severely prevented. In situ hybridization analysis using OSH1, a rice knotted1 (kn1) ortholog, confirmed the absence of lateral meristems in lax1-2 panicles. These defects indicate that the LAX1 gene is required for the initiation/maintenance of axillary meristems in the rice panicle. In addition to its role in forming lateral meristems, the wild-type LAX1 gene acts as a floral meristem identity gene which specifies the terminal spikelet meristem. A comparison of the defects in lax1-1 and lax1-2 plants suggested that the sensitivities to reduced LAX1 activity were not uniform among different types of meristems. In the fzp2 mutant panicle, the basic branching pattern of the panicle was indistinguishable from that of the wild type; however, specification of both terminal and lateral spikelet meristems was blocked, and sequential rounds of branching occurred at the point where the spikelet meristems are initiated in the wild-type panicle. This resulted in the generation of a panicle composed of excessive ramification of rachis-branches. The lax1-1 fzp2 double mutants exhibited a novel, basically additive, phenotype, which suggests that LAX1 and FZP2 function in genetically independent pathways.