Satellite sequence turnover in parthenogenetic systems: the apomictic triploid hybrid Bacillus lynceorum (Insecta, Phasmatodea)

In the genus Bacillus (Insecta, Phasmatodea) the Bag320 satellite DNA family is present in the bisexual B. grandii and in the related automictic nonhybrid B. atticus; it is lacking in the other bisexual taxon of the genus, B. rossius. This family of highly repeated sequences was analyzed for 11 populations of the apomictic triploid hybrid B. lynceorum. In the neighbor-joining dendrogram, B. lynceorum nucleotide sequences distribute, regardless of geographical origin, among two clusters, one also including all clones of the three B. atticus races, and the other including sequences of the B. grandii grandii subspecies. Thus, B. lynceorum is a trihybrid taxon: as the molecular approach definitively demonstrates, it embodies one haploid complement each of both B. grandii grandii and B. atticus, which must be added to that of B. rossius. The contribution of the latter species has already been assessed on karyological and allozymic grounds. A statistical analysis performed on p-distances shows that for the parental taxa, nucleotide substitution values are of comparable magnitudes at the population level but differ at the subspecific level, being higher for the bisexual taxon. In the apomictic hybrid, atticus- and grandii grandii- like sequences coexist with significantly different p-distance values. For three clones, the nucleotide compositions at the diagnostic loci suggest that gene conversion can occur between atticus- and grandii grandii-like monomers. On the whole, this supports bisexuality as a driving force in variant fixation and suggests that in Bacillus, different gametogenetic processes and different origins of the unisexuals are mirrored in genomic turnover rates of satellite DNA.      

The mitochondrial cytochrome oxidase II gene in Bacillus stick insects: ancestry of hybrids, androgenesis, and phylogenetic relationships

Sequencing of a cytochrome oxidase II (COII) gene fragment in Bacillus taxa provided evidence that the bisexual B. rossius is the maternal ancestor of the hybridogenetic B. rossius-grandii strains and revealed the same ancestry for both parthenogenetic hybrids: the diploid B. whitei (B. rossius/grandii grandii) and the triploid B. lynceorum (B. rossius/grandii grandii/atticus). Present data clearly demonstrate that all Bacillus unisexuals arose through asymmetrical hybridization events and realized a paraphyletic derivation from the B. rossius redtenbacheri subspecies. The invention of B. rossius mitochondrial DNA haplotypes in specimens with B. grandii grandii nuclear genomes revealed the occurrence of androgenesis in nature. Natural androgens represent a peculiar escape from hybridity and can help maintain the hybridogenetic system through the production of the fathering taxon via hybrid females. Results from the COII gene support the phyletic relationships among taxa suggested by previous taxonomical approaches, but also indicate a departure of B. grandii subspecies from the established taxonomy. Assuming the existence of a molecular clock, the evaluated substitution rate brings the splitting between B. rossius and B. grandii/B. atticus back to 22.79 +/- 2.65 myr before present, while the origin of hybrids appears to be much more recent (1.06 +/- 0.53 myr).     

New DAPI and FISH findings on egg maturation processes in related hybridogenetic and parthenogenetic Bacillus hybrids (Insecta, Phasmatodea)

Bacillus stick insects have proved adequate for studying a wide array of reproductive modes: sexual, parthenogenetic, hybridogenetic, androgenetic. Hybridogenetic strains (B. rossius-grandii) were thought to discard the paternal "grandii" haploset during first meiotic division and keep the "rossius" hemiclone, whereas the clonal B. whitei (=rossius/grandii) would maintain its hybrid structure by fusing back two nonsister nuclei-each derived from previously segregated heterospecific complements-by the end of the 2(nd) meiotic division. New investigations on laid eggs and ovariole squashes, either DAPI stained or FISH labeled, revealed that in hybridogens the "grandii" set is excluded from the germ line prior to meiosis and that a DNA extra-synthesis should occur to produce hemiclonal eggs after two cytologically normal meiotic divisions. On the other hand, in B. whitei eggs no genome segregation appears to occur and an intrameiotic DNA extra-synthesis must take place to produce 2n tetrachromatidic oocytes I; these divide twice and give unreduced clonal eggs. The new findings bring hybridogenetic oogenesis of Bacillus to be coincident with that of the known hemiclonal organisms and point to an independent onset of B. whitei from hemiclonal strains. In addition, B. whitei gains a closer resemblance to B. lynceorum owing to the sharing of a cytologically identical egg maturation mechanism, of the same maternal ancestor and of peculiar chromosomal features. It is here suggested that B. lynceorum originated from the incorporation of an "atticus" genome into a B. whitei egg, according to a pathway of repeated hybridization often occurred with other polyploid hybrids.     

Linkage between sexual and asexual lineages: genome evolution in Bacillus stick insects

The sexually reproducing stick insects Bacillus rossius and B. grandii are sharply differentiated in terms of allozyme gene alleles; B. atticus is a polyclonal automictic parthenogen sister to B. grandii grandii. Although well differentiated for coding genes, these hybridize to produce diploid (B. whitei=rossius/grandii) or triploid (B. lynceorum=rossius/grandii/atticus) clonal forms which reproduce apomictically. Allozyme analyses of unisexual Bacillus clearly establish their relationships from bisexual ancestor species as does the existence in all of them of several clones (especially in B. atticus) whose egg maturation allows regular recombination to occur. Bacillus taxa share the Bag320 satellite DNA family within different reproductive frameworks, allowing satellite variant homogenization to be uncoupled from fixation. The nested analysis of monomers reveals different patterns of sequence diversity: sexual reproduction includes both homogenization and variant fixation, whereas the slowing of molecular turnover processes and the absence of syngamy in the parthenogens realizes a similar range of sequence diversity at the level of the individual and supra‐individual, but with no fixation. On the other hand, the actual values of sequence diversity appear mostly linked to species traits – range size, copy number of repeats, number of hybrid crosses – and possibly transposon activity, rather than to the reproductive mode. In addition, the mitochondrial genome reveals a comparable level of cox2 sequence variability in sexual and parthenogenetic taxa, thus adding to clonal variability. From Bacillus and other stick insect complexes, an overall picture of genomic diversification of parthenogens is therefore beginning to emerge. To define those animals that reproduce by non‐canonical sexual modes (i.e. parthenogenesis, hybridogenesis), but make use of egg and meiotic mechanisms, the term meta‐sexual is proposed. © 2003 The Linnean Society of London. Biological Journal of the Linnean Society, 2003, 79 , 137–150.     

The Bag320 satellite DNA family in Bacillus stick insects (Phasmatodea): different rates of molecular evolution of highly repetitive DNA in bisexual and parthenogenic taxa

The Bag320 satellite DNA (satDNA) family was studied in seven populations of the stick insects Bacillus atticus (parthenogenetic, unisexual) and Bacillus grandii (bisexual). It was characterized as widespread in all zymoraces of B. atticus and in all subspecies of B. grandii. The copy number of this satellite is higher in the bisexual B. grandii (15%-20% of the genome) than in the parthenogenetic B. atticus (2%-5% of the genome). The nucleotide sequences of 12 Bag320 clones from B. atticus and 17 from B. grandii differed at 13 characteristic positions by fixed nucleotide substitutions. Thus, nucleotide sequences from both species cluster conspecifically in phylogenetic dendrograms. The nucleotide sequences derived from B. grandii grandii could be clearly discriminated from those of B. grandii benazzii and B. grandii maretimi on the basis of 25 variable sites, although all taxa come from Sicily. In contrast, the Bag320 sequences from B. atticus could not be discriminated accordingly, although they derive from geographically quite distant populations of its three zymoraces (the Italian and Greek B. atticus atticus, the Greek and Turkish B. atticus carius, and the Cyprian B. atticus cyprius). The different rate of evolutionary turnover of the Bag320 satDNA in both species can be related to their different modes of reproduction. This indicates that meiosis and chromosome segregation affect processes in satDNA diversification.      

Chromosomal and NOR patterns in the polyclonal stick insect Bacillus atticus atticus (Insecta; Phasmatodea).

Bacillus atticus atticus is a complex of thelytokous parthenogens, related to the bisexual Bacillus grandii, that ranges from Sardinia to Near Eastern countries. Karyotypic and cytogenetic differentiation of the B. atticus atticus diploid unisexual "isolates" is really higher than expected. Its standard karyotype has 2n = 34 chromosomes, but several instances of repatterned or even aneuploid complements have been found. The number and location of silver-stained NORs are particularly intriguing, since in addition to homozygous NOR patterns, simple or double hemizygous strains are found spread over specific and wide regions. The odd patterns are not due to Ag-NOR staining technique artifacts, since the FISH method, using rDNA probes, apparently labels the same ribosomal clusters. Transpositions and translocations have been suggested to account for some NOR patterns, but hybridizations between different NOR-bearing races are also a possible cause. This chromosomal survey clearly contributes to a better understanding of B. atticus phylogeny.     

Genome exclusion and gametic DAPI-DNA content in the hybridogenetic Bacillus rossius-grandii benazzii complex (Insecta Phasmatodea).

Among Sicilian stick insects, two hybridogenetic complexes have been discovered: Bacillus rossius-grandii benazzii and B. rossius-grandii grandii, which also produce androgenetic offspring. The egg maturation of the former is analyzed here through DAPI fluorometry, which, besides the assessment of the meiotic stages, also allows their DNA measurements and the analysis of sperm-head evolution into male pronuclei in these polyspermic eggs. Hybridogenetic eggs undergo an extrasynthesis of chromosomes, because two groups of n autobivalents (4C each) are segregated at metaphase 1st; the two groups must correspond to the pure parental species haplosets. Then the grandii chromosomes degenerate (1st polar body), while the rossius chromosomes divide further to produce two groups of n autodiads (2C each); one of them degenerates (2nd polar body), and the other is ready to perform syngamy (female pronucleus). Meanwhile, several B. grandii sperm evolve into male pronuclei by doubling their DNA (from 1C to 2C content) and assuming an interphase nucleus appearance. If regular mixis occurs, the F1 hybrid constitution is restored but, if it fails, a fusion between two sperms may occur, originating fully paternal descendants (natural androgenesis). The genome exclusion mechanism of stick-insect hybridogens appears to be more primitive than those observed in the already known hybridogenetic complexes of Poeciliopsis and Rana esculenta. Unfertilized eggs of hybridogens are capable of self-activation, but the cytology of the related clonally reproducing B. whitei indicates that its parthenogenetic mechanism stems from the hybridization event (hybrid theory) rather than from tychoparthenogenetic potentialities (spontaneous theory).     

HYBRIDOGENESIS AND ANDROGENESIS IN THE STICK-INSECT BACILLUS ROSSIUS-GRANDII BENAZZII (INSECTA,PHASMATODEA)

In northwestern Sicily interspecific hybrid females between Bacillus rossius and B. grandii benazzii (Insecta, Phasmatodea) are sympatric with facultatively parthenogenetic demes of the former and bisexual populations of the latter. Preliminary observations suggested that hybrid females are maintained by hybridogenetic reproduction, not by current F₁ hybrid production nor through parthenogenesis. Being hybridogens, a complex of hemiclonal lineages, we informally refer to them as B. rossius-grandii benazzii, according to Schultz's proposal. In this study B. rossius-g. benazzii females were crossed with males of B. g. benazzii, B. g. grandii, B. g. maretimi, and B. rossius. Allozyme analysis of the progeny showed that the great majority of them were actually produced by hybridogenesis with a hemiclonal inheritance of the maternal B. rossius genotype (Br^m) and actual syngamy with a sperm from the fathering male, so that Br^m-gb^p, Br^m-gg^p, Br^m-gm^p, and Br^m-r^p offspring were obtained in the respective crosses. All-paternal progeny (androgenetics) were also produced (Bgb^pgb^p, Bgm^pgm^p, Br^pr^p) and two gynogenetic descendants were observed. Cytological investigations on virgin eggs that failed to hatch revealed in most of them a haploid-diploid blocked blastoderm; this rudimentary parthenogenesis appears to be an important prerequisite for further evolution of this hybridogen. Reproductive modes of descendants were also analyzed; although Br^m-g^p hybrids are still able to reproduce by hybridogenesis, a progressive disruption of the hybridogenetic-androgenetic system takes place in synthetic B. rossius (Br^m-r^p, Br^pr^p) and abundant thelytokous parthenogenetic offspring are obtained from females of androgenetic origin. The evolutionary role of these hybridogens appears to be linked to their shift towards parthenogenesis; this has apparently occurred in the southeastern Sicilian hybrid B. whitei (=B. rossius/g. grandii), which exhibits both hybridogenesis and parthenogenesis.     

The library model for satellite DNA evolution: a case study with the rodents of the genus Ctenomys (Octodontidae) from the Iberá marsh, Argentina

On the basement of the library model of satellite DNA evolution is the differential amplification of subfamilies through lineages diversification. However, this idea has rarely been explored from an experimental point of view. In the present work, we analyzed copy number and sequence variability of RPCS (repetitive PvuII Ctenomys sequence), the major satellite DNA present in the genomes of the rodents of the genus Ctenomys, in a closely related group of species and forms inhabiting the Iberá marsh in Argentina. We studied the dependence of these two parameters at the intrapopulation level because in the case of interbreeding genomes, differences in RPCS copy number are due to recent amplification/contraction events. We found an inverse relationship among RPCS copy number and sequence variability: amplifications lead to a decrease in sequence variability, by means of biased homogenization of the overall satellite DNA, prevailing few variants. On the contrary, the contraction events that involve tandems of homogeneous monomers contribute-by default-minor variants to become "evident", which otherwise were undetectable. On the other hand, all the RPCS sequence variants are totally or partially shared by all the studied populations. As a whole, these results are comprehensible if these RPCS variants preexisted in the common ancestor of this Ctenomys group.     

Allozyme analysis and phyletic relationships of two new stick-insects from north-west Sicily: Bacillus grandii benazzii and B. rossius-grandii benazzii (Insecta Phasmatodea)

Two new Sicilian stick-insects have been discovered within the genus Bacillus. Evidence concerning the subspecific differentiation of Bacillus grandii benazzii and the hybrid constitution of B. rossius-grandii benazzii by means of allozyme analysis is given, and is consistent with morphological and karyological data on the differentiation of these two north-west Sicilian morphs from south-eastern B. g. grandii and B. whitei. B. g. benazzii is more polymorphic than B. g. grandii and B. rossius-g. benazzii embodies its genetic variability, thus differing sharply from the south-eastern parallel hybrid B. whitei ( = B. rossius × B. g. grandii). Reproductive mechanisms also appear to be different in the two interspecific hybrids, involving parthenogenesis in the latter and hybridogenesis in the former. Finally, the phyletic relationships among all Sicilian Bacillus taxa are summarized in a revised scheme, which also takes into account new evidence that B. atticus has contributed to hybrid constitution of the triploid B. lynceorum.     

Calamagrostis hakonensis (Poaceae): Distribution and differentiation of cytotypes

Chromosome counts were made for a total of 540 collections ofCalamagrostis hakonensis from 66 localities ranging from Kyushu to Hokkaido. Distribution and habitat preferences of the cytotypes involved are described. Sympatric occurrence of tetraploids (=semidiploids, 2n=28) and higher polyploids, as well as that of septaploids and plants at hexaploid and/or octoploid levels, was confirmed in several localities, and mixtures of hexaploids and octoploids within a population were frequently observed. Plants at hexaploid and octoploid levels were the most abundant and widespread. An examination of pollen of the voucher specimens showed that tetraploids (amphimictic) had good pollen, while higher polyploids (apomictic) were generally devoid of pollen and very rarely produced moderately good pollen, the grains of higher polyploids being larger than those of tetraploids. Distributions of tetraploids and higher polyploids were more precisely delineated through an examination of pollen of many herbarium specimens. In relation to the processes by which the complicated internal structure ofC. hakonensis has been established, the following subjects are discussed: infraspecific hybridization between ecotypically differentiated populations, enrichment of variability through occasional sexual reproduction expected in plants with more than 2n=42, persistence of the variants by apomictic reproduction, and probable roles of some extinct taxa.     

Chromosomal evidence of hemiclonal and all-paternal offspring production in Bacillus rossius-grandii benazzii (Insecta Phasmatodea)

The stick insects Bacillus rossius-grandii benazzii and B. rossius-grandii grandii naturally reproduce by hybridogenesis and androgenesis. The hybrid karyotype of the former (2n=35, XX female; 34, X0 male) clearly sums up a B. rossius haploset (r) with n=18 and a B. grandii benazzii one (gb) with n=17. The two sets keep the parental features for C-heterochromatin amount (much larger in the gb complement) and satellites/NORs (nuclear organizer regions) (more numerous and variably located in the r set); hybridogenetically produced males always show severely impaired gametogenesis and are therefore sterile, whereas hybridogenetic females are fertile. Reproductive, karyological and cytogenetical properties of the hybridogenetic system have been exploited to obtain the chromosomal evidence of whole haploset exchanges. In progenies obtained by crossing B. rossiusgrandii benazzii females to B. rossius males with either standard or repatterned (with Robertsonian fusions) karyotypes, there has always been complete agreement between electrophoretically genotyped and karyologically analyzed hybridogenetic offspring: the unassorted maternal r haploset (r ^m) is transmitted and the gb ^m haploset replaced by that of the fathering male (r ^p), thus evidencing the hemiclonal reproduction and the new r ^m-r ^p chromosomal constitution. New karyotype traits of the offspring relate to chromosome number (2n=36, female; 35, male), C-heterochromatin pattern (the heterochromatin-rich gb haploset completely disappears) and satellite/NOR features (corresponding to r ^m plus r ^p locations). The same crosses also produce genetically and chromosomally all-paternal descendants (androgenetics), of both sexes and fully fertile, with an r ^p r ^p structure. These androgenetic progeny show segregation for alleles and chromosomes at which fathering males are heterozygous: it was therefore possible to demonstrate that androgenetics can derive from syngamy of two sperm nuclei, of the several present in the polyspermic hybridogenetic egg. The production of androgenetics from field fertilized females of B. rossius-grandii benazzii, B. rossius-grandii grandii and parthenogenetic Bacillus whitei (=B. rossius/grandii grandii) suggests the occurrence of unsuspected relationships between hybrids and their parental species, so that the hybrids cannot be simply considered as sexual parasites. Furthermore, there is a suggestion of evolution of parthenogenetic clonal species from selection of initially hybridogenetic strains. The ability to produce uniparental progeny naturally from the spermatic genome may open a new field of investigation on genomic imprinting.     

Identification of differentially expressed cDNA sequences in ovaries of sexual and apomictic plants of Brachiaria brizantha

The isolation of genes associated with apomixis would improve understanding of the molecular mechanism of this mode of reproduction in plants as well as open the possibility of transfer of apomixis to sexual plants, enabling cloning of crops through seeds. Brachiaria brizantha is a highly apomictic grass species with 274 tetraploid apomicts accessions and only one diploid sexual. In this study we have compared gene expression in ovaries at megasporogenesis and megagametogenesis of sexual and apomictic accessions of B. brizantha by differential display (DD-PCR), with 60 primer combinations. Specificity of 65 cloned fragments, checked by reverse northern blot analysis, showed that 11 clones were differentially expressed, 6 in apomictic ovaries, 2 in sexual and 3 in apomictic and sexual, but at different stages. Of the 6 sequences isolated that were preferentially expressed in the apomictic accession: one sequence was from ovaries at megasporogenesis stage; three were from megagametogenesis stage; two were from both stages. Of the two sequences isolated from the sexual accessions, one showed expression in ovaries at megagametogenesis, while the other sequence was shown to be specific to both stages. Three sequences were from megasporogenesis stage in apomicts but were also detected at megagametogenesis in sexual plants. Sequence analysis showed that 5 of the 11 clones had no apparent homologues in the protein database. Some of the clones identified as apomictic-specific shared homology with known genes enabling their functional annotation. The relationships of these functions to the generation of the apomictic trait are discussed.     

Sexual reproduction development in apomictic Eulaliopsis binata (Poaceae)

Apomixis is a particular mode of reproduction that allows progeny formation without meiosis and fertilization. Eulaliopsis binata, a tetraploid apomictic species, is widely used for making paper, rope and mats. There is great potential for fixation of heterosis in E. binata due to autonomous endosperm formation in this species. Although most of its embryo sac originates from nucellus cells, termed apospory, we observed sexual reproduction initiation in 86.8 to 96.8% of the ovules, evidenced by callose deposition on the walls of cells undergoing megasporogenesis. However, only 2-3% mature polygonum-type sexual embryo sacs were confirmed by embryological investigation. Callose was not detected on aposporous initial cell walls. The aposporous initial cells differentiated during pre- and post-meiosis of the megaspore mother cell, while the sexual embryo sac degenerated at the megaspore stage. DNA content ratio of embryo and endosperm in some individuals was 2C:3C, based on flow cytometry screening of seed, similar to that of normal sexual seed. These results confirm that apomictic E. binata has conserved sexual reproduction to a certain degree, which may contribute to maintaining genetic diversity. The finding of sexual reproduction in apomictic E. binata could be useful for research on genetic mechanism of apomixis in E. binata.     

Sexual Hieracium pilosella plants are better inter-specific,while apomictic plants are better intra-specific competitors

Apomixis, asexual reproduction through seeds, occurs in over 40 plant families. This widespread phenomenon can lead to the fixation of successful genotypes, resulting in a fitness advantage. On the other hand, apomicts are expected to lose their fitness advantage if the environment changes because of their limited evolutionary potential, which is due to low genetic variability and the potential accumulation of deleterious somatic mutations. Nonetheless, some apomicts have been extremely successful, for example certain apomictic accessions of Hieracium pilosella L. from New Zealand, where the plant is invasive. Here, we investigate whether the success of these apomictic accessions could be due to a fitness advantage by comparing the vegetative competitiveness of apomictic H. pilosella from New Zealand with sexual accessions of H. pilosella from Europe. Sexual and apomictic plants were grown either (A) alone (no competition), (B) in competition with the other type (intra-specific competition), (C) in competition with the grass Bromus erectus (inter-specific competition), and (D) in competition with the other type and the grass B. erectus (intra- and inter-specific competition). To distinguish effects of apomixis and the region of origin, different H. pilosella lineages were compared. Furthermore, experiments were carried out to investigate effects of the ploidy level. We show that sexual plants are better inter-specific competitors than apomicts in terms of vegetative reproduction (number of stolons) and vegetative spread (stolon length), while apomicts do better than sexuals in intra-specific competition. The magnitude of the effect was in some cases dependent on the ploidy levels of the plants. Furthermore, apomicts always produced more stolons than sexuals, suggesting potential displacement of sexuals by apomicts where they co-occur.     

Reproductive strategy and population variability in the facultative apomict Hieracium pilosella (Asteraceae)

Molecular studies of apomictic plant species often detect more genotypic variation than predicted from their assumed reproductive mode. The two most commonly invoked mechanisms to explain these high levels of variation are recombination, via facultative sexuality, and mutation. The potential for sexual reproduction in the facultative apomict Hieracium pilosella (Asteraceae) was determined at three field sites by artificially pollinating with the closely related, but morphologically distinct, H. aurantiacum. The level of genotypic variation at the three sites was recorded using inter-simple sequence repeats (ISSRs). There was a significant, positive relationship between the measured potential for sexual reproduction and population genotypic variability, indicating that sex has played a role in the structuring of these populations; however, a causal relationship cannot be stated because of the use of regression. We also applied the recently developed method of compatibility analysis. Compatibility analysis can determine, using the occurrence of "character incompatibilities," whether patterns of variation observed in populations are most parsimoniously explained by mutation or recombination. Compatibility analysis also indicated that sexual reproduction had played a role in generating genotypic diversity in these populations. Combining these different types of data may give a greater understanding of the potential for the generation of genotypic diversity in facultative apomictic populations.     

Genetic tests of ancient asexuality in Root Knot Nematodes reveal recent hybrid origins

Background  

The existence of "ancient asexuals", taxa that have persisted for long periods of evolutionary history without sexual recombination, is both controversial and important for our understanding of the evolution and maintenance of sexual reproduction. A lack of sex has consequences not only for the ecology of the asexual organism but also for its genome. Several genetic signatures are predicted from long-term asexual (apomictic) reproduction including (i) large "allelic" sequence divergence (ii) lack of phylogenetic clustering of "alleles" within morphological species and (iii) decay and loss of genes specific to meiosis and sexual reproduction. These genetic signatures can be hard to assess since it is difficult to demonstrate the allelic nature of very divergent sequences, divergence levels may be complicated by processes such as inter-specific hybridization, and genes may have secondary roles unrelated to sexual reproduction. Apomictic species of Meloidogyne root knot nematodes have been suggested previously to be ancient asexuals. Their relatives reproduce by meiotic parthenogenesis or facultative sexuality, which in combination with the abundance of nematode genomic sequence data, makes them a powerful system in which to study the consequences of reproductive mode on genomic divergence.     

The Pattern of Genetic Variability in Apomictic Clones of Taraxacum officinale Indicates the Alternation of Asexual and Sexual Histories of Apomicts

Dandelions (genus Taraxacum) comprise a group of sexual diploids and apomictic polyploids with a complicated reticular evolution. Apomixis (clonal reproduction through seeds) in this genus is considered to be obligate, and therefore represent a good model for studying the role of asexual reproduction in microevolutionary processes of apomictic genera. In our study, a total of 187 apomictic individuals composing a set of nine microspecies (sampled across wide geographic area in Europe) were genotyped for six microsatellite loci and for 162 amplified fragment length polymorphism (AFLP) markers. Our results indicated that significant genetic similarity existed within accessions with low numbers of genotypes. Genotypic variability was high among accessions but low within accessions. Clustering methods discriminated individuals into nine groups corresponding to their phenotypes. Furthermore, two groups of apomictic genotypes were observed, which suggests that they had different asexual histories. A matrix compatibility test suggests that most of the variability within accession groups was mutational in origin. However, the presence of recombination was also detected. The accumulation of mutations in asexual clones leads to the establishment of a network of clone mates. However, this study suggests that the clones primarily originated from the hybridisation between sexual and apomicts.     

Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha

Background  

Brachiaria brizantha is an important forage grass. The occurrence of both apomictic and sexual reproduction within Brachiaria makes it an interesting system for understanding the molecular pathways involved in both modes of reproduction. Quantitative real time PCR (qRT-PCR) has emerged as an important technique to compare expression profile of target genes and, in order to obtain reliable results, it is important to have suitable reference genes. In this work, we evaluated eight potential reference genes for B. brizantha qRT-PCR experiments, isolated from cDNA ovary libraries. Vegetative and reproductive tissues of apomictic and sexual B. brizantha were tested to validate the reference genes, including the female gametophyte, where differences in the expression profile between sexual and apomictic plants must occur.