Wolbachia induced cytogenetical effects as evidenced in Chorthippus parallelus (Orthoptera).

The cytoplasmic incompatibility induced by the bacterial endosymbiont Wolbachia is attributed to chromatin modification in the sperm of infected individuals and is only 'rescued' by infected females after fertilization. Chorthippus parallelus is a grasshopper with 2 subspecies that form a hybrid zone in the Pyrenees in which this Wolbachia-generated cytoplasmic incompatibility has recently been described. The analysis of certain cytogenetic traits (sex chromosome-linked heterochromatic bands, nucleolar organizing region expression, spermatid size and morphology, and number of chiasmata formed) in pure and hybrid Chorthippus parallelus that are infected and not infected by this bacterium indicates that the infection affects some of these traits and, in the case of the spermatids, reveals a synergism between the infection and the hybrid condition. These results are interpreted as being secondary effects of the chromatin modification induced by Wolbachia which thereby support this model of modification/rescue. The possible effects of these cytogenetic variations on affected individuals are also considered.

Identification of genes specifically expressed in cauliflower reproductive meristems. Molecular characterization of BoREM1.

Using the meristems of the cauliflower curd as a source of tissue and a series of subtractive hybridizations and amplification reactions, we have constructed a cDNA library highly enriched in cDNAs expressed in reproductive meristems. The analysis of a sample of 250 clones from this library identified 22 cDNA clones corresponding to genes specifically expressed in these cauliflower meristems. Apart from two clones that corresponded to APETALA1, and two other ones showing similarity to different aminoacyl-tRNA synthetases, the remaining clones showed no similarity to any sequence in the databases and may correspond to novel genes. One of these clones, BoREM1, was further characterized and found to correspond to a gene encoding a protein with features of regulatory proteins that follows a expression pattern very similar to the LEAFY transcripts.

Relationship between the levels of wheat-rye metaphase I chromosomal pairing and recombination revealed by GISH.

The metaphase I and anaphase I stages of meiosis of wheat x rye hybrids carrying the ph1b mutation were analyzed by genomic in situ hybridization. This technique allows distinction between three different types of wheat-rye associations in metaphase I configurations as well as detection of wheat-rye recombinant chromosomes in anaphase I cells. The frequency of associations between wheat and rye chromosomes greatly exceeded the level of wheat-rye recombination found in the three hybrids examined. Extremely distal associations, which account for about 50% of the total wheat-rye metaphase I chromosomal pairing, can explain such a discrepancy between metaphase I and anaphase I data. It is further discussed whether these associations reflect very distally located chiasmata or nonchiasmatic pairing. The sizes of the segments exchanged in wheat-rye recombinant chromosomes provide cytological evidence that wheat-rye recombination is restricted to the distal chromosomal regions.

Equilocality and heterogeneity of constitutive heterochromatin: in situ localization of two families of highly repetitive DNA in Dociostaurus genei (Orthoptera).

The chromosome complement of the grasshopper Dociostaurus genei is characterized by the presence of constitutive heterochromatin (C-bands) located in the centromeric regions of all the chromosomes and in the distal regions of some autosomes in the form of supernumerary segments. A sequence analysis was carried out to obtain information about the molecular characteristics of both heterochromatic regions. Two families of tandemly repetitive DNA (DgT2 and DgA3) from D. genei were cloned and characterized. Data obtained from in situ hybridization indicate that these families are located solely in the regions of constitutive heterochromatin. The DgT2 clone is representative of a family of sequences which mainly forms the centromeric C-bands in each chromosome of the complement. The DgA3 family is the major component of the distal C-bands (supernumerary segments) present in most of the autosomal pairs. These results show the existence in D. genei of two different families of repetitive DNA restricted to different chromosomal domains. We discuss these results in the light of the possible role of chromosomal disposition in the maintenance of the differences between heterochromatic DNA from different chromosomal regions and the homogenization of DNA sequences from equilocal chromosomal domains.

Meiotic pairing in wheat-rye derivatives detected by genomic in situ hybridization and C-banding--A comparative analysis.

Meiosis of triticalextetraploid rye hybrids (genome constitution ABRRR) was analysed by genomic in situ hybridization (GISH) and C-banding. The results obtained reveal a considerable difference between these techniques with regard to their efficiency in detecting any type of pairing, either homologous or homoeologous. Thus the percentage of pollen mother cells containing wheat/rye homoeologous associations determined by C-banding and GISH was 2.5 and 9.2, respectively. Such a discrepancy can be ascribed to a certain proportion of wheat/rye associations not being identified by C-banding. The potential and limitations of the two techniques for meiotic analysis are discussed.

Metaphase I-bound arms frequency and genome analysis in wheat-Aegilops hybrids. 3. Similar relationships between the B genome of wheat and S or S (l) genomes of Ae. speltoides, Ae. longissima and Ae. sharonensis.

The meiotic behaviour of Triticum aestivum × Aegilops speltoides, T. aestivum × Ae. sharonensis and T. aestivum × Ae. longissima tetraploid hybrids (genome constitution ABDS, ABDS (l) , and ABDS (l) , respectively) has been analysed by the C-banding technique. Of the six types of pairing normally occurring, at metaphase I three were recognized: A-D, AD-BS/AD-BS (l) and B-S/B-S (l) . The relative order observed in the low pairing hybrid, A-D> B-S (l) >AD-BS (l) , as well as that found in high-pairing 'Chinese Spring' × Ae. speltoides hybrids, A-D>AD-BS>ß-S, revealed the existence of preferential pairing patterns among the different genomes that are in competition. In all of the hybrids analysed the mean number of bound arms per cell for the A-D type was significantly higher than the mean number of associations between the B and S/S (l) genomes. Usually the relative contribution of each type of pairing is maintained among hybrids with different Aegilops species. These results indicate that the genomes of Ae. speltoides, Ae. sharonensis and Ae. longissima show a similar affinity with the genomes of hexaploid wheat; therefore none of these species can be considered to be a distinct donor of the B genome of wheats.

Metaphase-I bound-arm frequency and genome analysis in wheat-Aegilops hybrids. 2. Cytogenetical evidence for excluding Ae. Sharonensis as the donor of the B genome of polyploid wheats.

Genome affinities were analyzed at meiosis in C-banded metaphase-I cells of wheat x Ae. Sharonensis hybrid plants. The results showed that the most frequent type of pairing occurred between chromosomes of the A and D genomes in all plants, as well as in cells with different numbers of associations. These findings clearly indicated that Ae. Sharonensis can be excluded as the donor of the B genome of wheat.

Mapping of genes controlling seed storage-proteins and cytological markers on chromosome 1R of rye.

Rye secalins, telomeric C-bands, and telocentric chromosomes were used as markers in the progeny of a test-cross in order to determine the position of seed storage-protein genes Glu-R1 and Gli-R1 with respect to the centromere and both telomeres of chromosome 1 R in rye. These genes were linked to the centromere (32.35±3.28% and 36.27±3.37% recombination, respectively). Glu-R1 was loosely linked to the telomere of the long arm (43.63±3.47% recombination), while Gli-R1 was closely linked to the telomere of the short arm (9.80±2.08% recombination). This finding supports the possibility that rye ω- and γ-secalin genes may be located on the satellites, as has been described in wheat for genes that code similar proteins. The importance of metaphase-I pairing failure and its consequences for the estimation of the recombination fraction are also discussed.

Metaphase I bound arms frequency and genome analysis in wheat-Aegilops hybrids : 1. Ae. variabilis-wheat and Ae. kotschyi-wheat hybrids with low and high homoeologous pairing.

Meiotic associations of different wheat-Aegilops variabilis and wheat-Ae. kotschyi hybrid combinations with low and high homoeologous pairing were analyzed at metaphase I. Five types of pairing involving wheat and Aegilops genomes were identified by using C-banding. A genotype that seems to promote homoeologous pairing has been found in Ae. variabilis var. cylindrostachys. Its effect is detectable in the low pairing hybrids but not in the high ones. Pairing affinity has been analyzed on the basis of metaphase I associations in the low and high homoeologous pairing hybrids, and in bivalents and multivalents in the high pairing hybrids. The results indicate that the amount of bound arms of each type of identifiable association relative to the total associations formed (relative contribution) was not maintained, either between the different levels of pairing (low and high) or between different meiotic configurations (bivalents and multivalents). These findings seem to indicate that quantifications of genomic relationships based on the amount of chromosome pairing at metaphase I must be carefully done in this type of hybrid combinations.