BACKGROUND AND AIMS The tam (tardy asynchronous meiosis) mutant of Arabidopsis thaliana, which exhibits a modified cytokinesis with a switch from simultaneous to successive cytokinesis, was used to perform a direct test of the implication of cytokinesis in aperture-pattern ontogeny of angiosperm pollen grains. The aperture pattern corresponds to the number and arrangement of apertures (areas of the pollen wall permitting pollen tube germination) on the surface of the pollen grain. METHODS A comparative analysis of meiosis and aperture distribution was performed in two mutant strains of arabidopsis: quartet and quartet-tam. KEY RESULTS While the number of apertures is not affected in the quartet-tam mutant, the arrangement of the three apertures is modified compared with the quartet, resulting in a different aperture pattern. CONCLUSIONS These results directly demonstrate the relationship between the type of sporocytic cytokinesis and pollen aperture-pattern ontogeny.

We examined large-scale patterns of morphology, genetic structure and ecological correlates of Fraxinus excelsior and the closely related species Fraxinus angustifolia in France, in order to determine the degree of hybridization between them. We sampled 24 populations in two putative hybrid zones (Loire and Saône), and five control populations of each species. We measured foliar characteristics of adult trees and used five nuclear microsatellites as molecular markers. Canonical discriminant analysis indicated that the two species differ in morphology, but that intermediate types are common in the Loire region but less frequent in the Saône region. Bayesian population assignment identified one F. angustifolia and two F. excelsior gene pools. Most Loire individuals clustered genetically with the F. angustifolia gene pool. In contrast, the Saône region presented individuals belonging mostly to F. excelsior pools, although the F. angustifolia type was frequent in certain populations. The lowest F(ST) values were found between the Loire and F. angustifolia controls that also exhibited no significant isolation by distance. The proportion of the F. angustifolia gene pool in each locality was negatively correlated with winter temperatures, suggesting that a cold climate may be limiting. Hybridization is probably favoured by the intermediate climatic conditions in the Loire region that allow both species to occur, but is somewhat hampered by the harsher winters in the Saône area where morphological introgression has apparently not yet occurred.

The study of how parasites adapt to new hosts is of great importance for understanding the emergence of new diseases. Here, we report a study of the anther smut disease on Gypsophila repens (Caryophyllaceae). In contrast to what is usually found on other host species, infected natural populations of G. repens are extremely rare. Moreover, symptoms of diseased plants are incomplete and highly variable over the time. These results suggest that the fungus infecting G. repens is a case of a parasite not capable of exploiting its host optimally. Molecular analyses of Microbotryum violaceum strains infecting this and other Caryophyllaceae revealed that this sub-optimal behaviour probably resulted from a recent host shift from the morphologically similar plant Petrorhagia saxifraga. With its exceptionally low virulence and prevalence, but apparent self-sustainability, the disease on G. repens may thus represent an interesting case study for investigating the conditions leading to adaptation of parasites on new hosts.

In eudicot postmeiotic tetrads, apertures are usually joined in pairs in highly conserved areas. These appear to be located at the last points of contact persisting at the end of cytokinesis between the cytoplasm of the future microspores. In order to investigate the relationship between cytokinesis and aperture formation, aperture distribution within postmeiotic tetrads and the progression of meiosis were studied in Nicotiana tabacum cv. Ambalema. This variety (inbred line) produces about 85% tricolporate pollen and 15% tetracolporate pollen grains. In addition, about 7% of tetrads are composed of four equal-sized microspores and a supernumerary pseudomicrospore of small size and an equal proportion of tetrads exhibit unpaired apertures (these apertures are not joined in pairs within tetrads). Observation of cytokinesis indicates that both unpaired apertures and pseudomicrospores could result from the persistence of late communications between microsporocytes. Observations of tetrads indicate that an increase in the number of elements that are separated during cytokinesis is correlated with an increase in microspore aperture number. All data converge to support the hypothesis that aperture site determination is partly controlled by the number of walls formed to separate the different elements of the tetrad.

Restriction fragment length polymorphism, polymerase chain reaction-restriction fragment length polymorphism and simple sequence repeat (SSR or microsatellites) analyses were performed to detect chloroplast DNA polymorphisms between two ash species, Fraxinus excelsior and F. angustifolia. Only one SSR locus was found to be polymorphic, confirming the very close relatedness of these species. Inheritance of this marker was studied in hybrids obtained from controlled crosses between the two tree species. Results indicated, for the first time in Oleaceae, that chloroplasts are maternally inherited. This chloroplast SSR marker is now used concomitantly with nuclear markers to analyse ash populations in sympatric areas.

In most flowering plants, a single cytokinesis follows the two meiotic divisions during pollen-grain ontogeny. Aperture pattern (i.e., aperture number and distribution on pollen surface) ontogeny could be linked to the processes ensuring the apportionment of the cytoplasm to the four microspores.This apportionment is achieved by radial arrays of microtubules organized around the nuclei. The cleavage planes are defined in the overlapping regions of opposing arrays extending from different nuclei. We followed the establishment of these arrays in two different lines of plants belonging to the genus Nicotiana that produce pollen grains with different aperture numbers. Different distributions of the microtubules have been observed, which can be interpreted as resulting from variation in the interactions between nuclei; these distributions appear to be correlated with aperture number.As a consequence, we propose that simultaneous cytokinesis allows the formation of multiple pollen morphologies. This mechanism is consistent with aperture number distribution within angiosperms and provides clues to help our understanding of the evolution of aperture number.

Three different dissection treatments were applied to mature seeds of two European ash species: Fraxinus excelsior(L.) and Fraxinus angustifolia(Vahl.) in order to compare their relative efficiency for germination and further development. The in vitro embryo culture appeared to be the most efficient for both species (nearly 90% of viable plants). The two species expressed differences in germi- nation rate without embryo culture.

Pollen grains display a wide range of variation in aperture number and arrangement (pattern) in angiosperms. Apertures are well-defined areas of the pollen wall surface that permit pollen tube germination. For low aperture numbers, aperture patterns are characteristic of the major taxonomic divisions of angiosperms. This paper presents a developmental model that explains most of the aperture patterns that are recorded in angiosperms. It is based on the analysis of the different events that occur during meiosis and lead to microspore differentiation. It demonstrates that variation occurring during meiosis in angiosperms is sufficient to produce the core morphological set of the most commonly observed pollen morphologies.

A simplified method for the isolation of mitochondrial DNA (mtDNA) from several angiosperm tree species is described. The procedure does not require gradient ultracentrifugation or organic solvent extractions and can be performed in a single day. mtDNA was isolated from flower buds collected from adult trees just after the first pollen mitosis and purified by gravity flow through disposable columns containing an anion-exchange resin. The method led to restrictable mtDNA from the tree species analysed. Restriction patterns enabled us to distinguish between European and American ashes.

Individuals of Arabidopsis thaliana, collected in different natural populations, were grown in controlled and elevated CO2 in a glasshouse. Following germination, root growth of progeny of different lines of these populations was studied in control and elevated atmospheric CO2. No significant direct effect of atmospheric CO2 concentration could be demonstrated on root growth. An important parental effect was apparent, namely that root length and branching were decreased in seeds collected from a mother plant which had been grown in elevated CO2. This was correlated with smaller seeds, containing less nitrogen. These parental effects were genetically variable. We conclude that CO2 may affect plant fitness via parental effects on seed size and early root growth and that the genetic variability shown in our study demonstrates that Arabidopsis populations will evolve in the face of this new selective pressure. (C) 1998 Annals of Botany Company.

Ribosomal DNA polymorphisms were studied in 6 lines of Petunia hybrida using EcoRI, BamHI, HindIII, KpnI, Sad or XhoI. Each line carries several unit types, and 13 types were found in lines, which was not expected. We characterized the unit types and we determined the number of loci. Two kinds of unit types carrying no or several HindIII sites were revealed. The longest EcoRI and BamHI fragments in St43 correspond to a 11.4 kb unit type. Moreover, a 2.6 kb EcoRI fragment cannot be mapped in the 11.4 kb unit. It was found to be equivalent to the 2.45 kb EcoRI fragment carrying the 25 S rRNA coding sequence. Consequently, it was mapped in another unit 11.7 kb long. In Tlh1 the corresponding EcoRI and BamHI fragments enabled us to construct 8.8, 9.2 and 10.8 kb segments. These fragments are therefore considered to be part of the 11.4 kb unit length. Other lines display combinations of these length units. The inheritance of polymorphic fragments of lines (St43 and Tlh1) for 50 individuals of the 2 possible backcrosses [(St43 x Tlh1) x St43] and [(St43 x Tlh1) x Tlh1] indicated at least 2 loci. The presence in Sk176 of 6.2, 5.7 and 5.4 EcoRI fragments suggested 3 loci. The haploid plants obtained from the hybrid (St43 x Tlh1) display 1 individual carrying the 3 unit types present in the hybrid which proves the presence of 3 rDNA loci in Tlh1. Moreover, the segregation in the backcrosses corresponds to only 2 loci in St43. It carries a nulli-allele. Evidence for such hypotheses were obtained by in situ hybridization with a biotinylated probe. The Tlh1 and Tlh7 dihaploid lines display more unit types and, consequently, more polymorphisms than other lines.

The ovaries of two different Petunia species: Petunia hybrida (hort) and Petunia parodii (Steere) were irradiated with τ-ray doses ranging from 50 to 1,000 Gy before pollination. Seed setting occurred after 4 days preculture on a non-sterile medium. Ovaries transformed into fruits were then cultivated aseptically with the following results: (1) τ-ray doses ranging from 200 to 1,000 Gy led to the development of two types of plants: haploids 2n=x=7 and overdiploids 2n>2x=14. (2) The androgenetic origin of haploids was ascertained by using genetic markers. The origin of overdiploids is discussed. (3) Androgenetic haploids contained the chloroplasts of the irradiated female parent. No visible change of cp DNA patterns was observed after irradiation. (4) The four possible androgenetic events were successfully obtained between the two Petunia species: hybrida haploids with hybrida or parodii cytoplasm, and parodii haploids with parodii or hybrida cytoplasm.

Various sugars (sucrose, lactose, maltose, glucose, fructose and galactose) were tested as carbon sources for in vitro anther cultures of Petunia, with the following results:

the yield of androgenetic plants is conditionned in Petunia by two important distinct factors, i.e., yield of embryos and quality of these same embryos.