Polyploidization is a widespread process that results in the merger of two or more genomes in a common nucleus. To investigate modifications of gene expression occurring during allopolyploid formation, the Brassica napus allotetraploid model was chosen. Large-scale analyses of the proteome were conducted on two organs, the stem and root, so that .1600 polypeptides were screened. Comparative proteomics of synthetic B. napus and its homozygous diploid progenitors B. rapa and B. oleracea showed that very few proteins disappeared or appeared in the amphiploids (,1%), but a strikingly high number (25-38%) of polypeptides displayed quantitative nonadditive pattern. Nonstochastic gene expression repatterning was found since 99% of the detected variations were reproducible in four independently created amphiploids. More than 60% of proteins displayed a nonadditive pattern closer to the paternal parent B. rapa. Interspecific hybridization triggered the majority of the deviations (89%), whereas very few variations ($3%) were associated with genome doubling and more significant alterations arose from selfing ($9%). Some nonadditive proteins behaved similarly in both organs, while others exhibited contrasted behavior, showing rapid organ-specific regulation. B. napus formation was therefore correlated with immediate and directed nonadditive changes in gene expression, suggesting that the early steps of allopolyploidization repatterning are controlled by nonstochastic mechanisms.
Summary• Polyploidy, or whole genome duplication, is a major evolutionary process that has shaped eukaryotic genomes, notably those of flowering plants. The mechanisms underlying the regulation of, and sharing of functions between, the duplicated genes originating from polyploidy events, which lead to novel phenotypes, remain to be elucidated.• A previous comparative proteomic study identified 360 proteins that were differentially regulated between the diploid Brassica progenitors and their synthetic allotetraploid derivatives. For 102 of these proteins, using the same resynthesized Brassica napus allotetraploids, we assayed the accumulation of the transcripts of the corresponding genes. We compared transcript levels quantified in the synthetic allotetraploids with the mid-parent expression values.• Although all of the genes surveyed encoded nonadditive proteins, we found that two-thirds of them had additive transcript levels, indicating that most of the differential protein regulation is not explained by transcriptional changes.• Our data suggest that differential protein regulation is mainly governed by posttranscriptional modifications. Summarizing available data from transcriptomic studies of other synthetic allopolyploid models, we describe the general trends of transcript regulation in an allopolyploid genome and discuss putative underlying molecular mechanisms, with particular emphasis on the small RNA pathway for the post-transcriptional control of gene expression.
Polyploidization is a major evolutionary process in eukaryotes. In plants, genetic and epigenetic changes occur rapidly after formation of allopolyploids. Hybridization, rather than genome doubling itself, is considered as the main cause for the resulting differential gene expression. We studied the consequences of genome doubling alone in an autopolyploid model, by comparing two-dimensional gel electrophoresis (2-DE) gels of haploid, diploid, and tetraploid Brassica oleracea cabbages. Two fully homozygous lines, HDEM and RC, as well as two organs, leaf and stem, were studied. For the 558 common spots found present in all the 29 2-DE gels of the experiment, inter-organ and -genotype differences were the major sources of the variation in protein amounts: 41 and 10-13%, respectively. HDEM leaf and stem proteomes were not significantly affected by the ploidy level, since no qualitative variation was detected and since the number of quantitative variations could be due to chance. For RC, no qualitative variations were observed, but a few spots were significantly variable in protein amount. However, the number of inter-ploidy variations was of the same range as the number of intra-ploidy variations. In conclusion, whatever the ploidy level, leaf and stem proteomes remained globally unchanged in both cabbage lines.
SummaryThe role played by whole-genome duplication (WGD) in evolution and adaptation is particularly well illustrated in allopolyploids, where WGD is concomitant with interspecific hybridization. This 'Genome Shock', usually accompanied by structural and functional modifications, has been associated with the activation of transposable elements (TEs). However, the impact of allopolyploidy on TEs has been studied in only a few polyploid species, and not in Brassica, which has been marked by recurrent polyploidy events.Here, we developed sequence-specific amplification polymorphism (SSAP) markers for three contrasting TEs, and compared profiles between resynthesized Brassica napus allotetraploids and their diploid Brassica progenitors. To evaluate restructuring at TE insertion sites, we scored changes in SSAP profiles and analysed a large set of differentially amplified SSAP bands.No massive structural changes associated with the three TEs surveyed were detected. However, several transposition events, specific to the youngest TE originating from the B. oleracea genome, were identified.Our study supports the hypothesis that TE responses to allopolyploidy are highly specific. The changes observed in SSAP profiles lead us to hypothesize that they may partly result from changes in DNA methylation, questioning the role of epigenetics during the formation of a new allopolyploid genome.
Background: Allopolyploidy is a preeminent process in plant evolution that results from the merger of distinct genomes in a common nucleus via inter-specific hybridization. Allopolyploid formation is usually related to genome-wide structural and functional changes though the underlying mechanisms operating during this "genomic shock" still remain poorly known. The aim of the present study was to investigate the modifications occurring at the proteomic level following an allopolyploidization event and to determine whether these changes are related to functional properties of the proteins. In a previous report, we applied comparative proteomics to synthetic amphiploids of Brassica napus and to its diploid progenitors B. rapa and B. oleracea. Although several hundred polypeptides displayed additivity (i.e. mid-parent values) in the amphiploids, many of them showed non-additivity. Here, we report the in silico functional characterization of the "non-additive" proteins (the ones with a non-additive pattern of regulation) in synthetic B. napus.
Several statistical methods were investigated for improving yield estimation for 72 genotypes of winter wheat tested in a single-row-plot (SRP) trial. The different analytical models were compared using several criteria: the residual mean square fit; the standard error of differences of genotype means; the correlations of genotype means with means from sixrow-plot (6RP) reference trial; the expected genetic gains relative to 6RP trial from selecting the 10 top yielding genotypes.There was no significant correlation between the unadjusted genotype means for SRP and 6RP. Genotype yields were positively correlated with height in SRP, but this correlation was reversed in 6RP. Using the mean height difference of a plot with its two neighbours as a covariate to adjust for competition in SRP reduced the standard error of difference of genotype means and substantially increased their correlation with the 6RP means. The predicted selection efficiency in SRP trial increased from zero, before adjustment, to 83 % of 6RP trial. Including the difference in heading date and the effect of guards at borders as covariates in the SRP analysis improved the residual mean square fit but had no effect on correlation with 6RP means or selection efficiency. A producer-competitor model in which individual competitor effects were estimated for each genotype gave a substantial improvement in fit over the covariate models but a much lower correlation with 6RP means and selection efficiency.Yield selection in early generation wheat trials based on single-row plots could be considerably strengthened by use of a plot covariate derived from height difference between neighbours.
Outcrossing rates within the wild green foxtail, Setaria viridis, and the cultivated foxtail millet, S. italica, are very low. However, spontaneous interspecific hybridizations in the experimental garden occurred in both directions at rates ranging from 0.002% to 0.6% according to plant density and distance between parents. Offtypes found in farmers' fields where foxtail millet is cultivated were shown to have originated from such interspecific crosses. Differences in the EcoR1 patterns of chloroplast DNA between cultivated and wild plants indicated that reciprocal crosses do occur in the field. These findings indicate that even a largely selfing cultivated species may exchange genetic information with wild relatives at rates that may cause problems if transgenic cultivars are released.
Strong selection within a given population locally reduces genetic variability not only in the selected gene itself but also in neighbouring loci. This so-called hitch-hiking effect is related to the initial linkage disequilibrium between markers and the selected gene, and depends mainly on the number of copies of the beneficial allele at the start of the selection phase. Contrary to the classical case, in which selection acts on a single, newly arisen beneficial mutation, we considered selection from standing variation (soft selective sweeps) on a gene (Rht-B1) with a major effect on plant height, a selected trait in an experimental wheat population grown for 17 generations, and we documented the evolution of gene diversity and linkage disequilibrium near this gene. As expected, Rht-B1 was found to be under strong selection (s = 0.15) and its variation in frequency accounted for 15% of the total trait evolution. This led to a smaller genetic effective population size at Rht-B1 (N(eg) = 18) compared to the whole genome estimation (N(eg) = 167). When compared with expectations under genetic drift only, no significant decrease in gene diversity was found at the closest loci. We computed expected di-locus frequencies for any linked marker-Rht-B1 pair due to hitch-hiking effects. We found that hitch-hiking was expected to affect the two most closely linked loci, but expected reduction in gene diversity was not greater than that due to genetic drift, which was consistent with the observations. Such limited effect was attributed to the low level of linkage disequilibrium (0.16) estimated after parental intercrosses, together with a relatively high initial frequency of the gene. This situation is favourable to candidate gene approaches where small linkage disequilibrium around selected genes is expected.
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