Wheat alleles introgress into selfing wild relatives: empirical estimates from approximate Bayesian computation in Aegilops triuncialis

Extensive gene flow between wheat (Triticum sp.) and several wild relatives of the genus Aegilops has recently been detected despite notoriously high levels of selfing in these species. Here, we assess and model the spread of wheat alleles into natural populations of the barbed goatgrass (Aegilops triuncialis), a wild wheat relative prevailing in the Mediterranean flora. Our sampling, based on an extensive survey of 31 Ae. triuncialis populations collected along a 60 km × 20 km area in southern Spain (Grazalema Mountain chain, Andalousia, totalling 458 specimens), is completed with 33 wheat cultivars representative of the European domesticated pool. All specimens were genotyped with amplified fragment length polymorphism with the aim of estimating wheat admixture levels in Ae. triuncialis populations. This survey first confirmed extensive hybridization and backcrossing of wheat into the wild species. We then used explicit modelling of populations and approximate Bayesian computation to estimate the selfing rate of Ae. triuncialis along with the magnitude, the tempo and the geographical distance over which wheat alleles introgress into Ae. triuncialis populations. These simulations confirmed that extensive introgression of wheat alleles (2.7 × 10^?4 wheat immigrants for each Ae. triuncialis resident, at each generation) into Ae. triuncialis occurs despite a high selfing rate (Fis ≈ 1 and selfing rate = 97%). These results are discussed in the light of risks associated with the release of genetically modified wheat cultivars in Mediterranean agrosystems.     

Spatial and temporal genetic dynamics of the grasshopper Oedaleus decorus revealed by museum genomics

Analyzing genetic variation through time and space is important to identify key evolutionary and ecological processes in populations. However, using contemporary genetic data to infer the dynamics of genetic diversity may be at risk of a bias, as inferences are performed from a set of extant populations, setting aside unavailable, rare, or now extinct lineages. Here, we took advantage of new developments in next‐generation sequencing to analyze the spatial and temporal genetic dynamics of the grasshopper Oedaleus decorus, a steppic Southwestern‐Palearctic species. We applied a recently developed hybridization capture (hyRAD) protocol that allows retrieving orthologous sequences even from degraded DNA characteristic of museum specimens. We identified single nucleotide polymorphisms in 68 historical and 51 modern samples in order to (i) unravel the spatial genetic structure across part of the species distribution and (ii) assess the loss of genetic diversity over the past century in Swiss populations. Our results revealed (i) the presence of three potential glacial refugia spread across the European continent and converging spatially in the Alpine area. In addition, and despite a limited population sample size, our results indicate (ii) a loss of allelic richness in contemporary Swiss populations compared to historical populations, whereas levels of expected heterozygosities were not significantly different. This observation is compatible with an increase in the bottleneck magnitude experienced by central European populations of O. decorus following human‐mediated land‐use change impacting steppic habitats. Our results confirm that application of hyRAD to museum samples produces valuable information to study genetic processes across time and space.     

Measuring host finding capacity and arrestment of natural enemies of the cassava mealybug,Phenacoccus manihoti, in the field

In unifested fields, 80 cassava tips were artificially infested with 0, 1, 2, 4, 8, 16, 32, or 64 third instars, and 20 or 100 eggs of cassava mealybug,Phenacoccus manihoti Matile-Ferrero (Hom., Pseudococcidae). Another 80 uninfested tips served as a control. Tips were arranged in a circle of 28 m diameter, in the centre of which the following exotic natural enemies ofP. manihoti were released:Apoanagyrus (Epidinocarsis) lopezi De Santis andA. diversicornis (Howard) (Hym., Encyrtidae),Hyperaspis notata (Mulsant) andDiomus hennesseyi Fürsch (Col., Coccinellidae), and others. This experiment was repeated six times. During the 4–14 days following release, all experimental tips were inspected at two-hour intervals during each day and the presence of exotic as well as indigenous natural enemies, likeExochomus troberti Mulsant (Col., Coccinellidae), ants and spiders was noted. The experiment was repeated six times measured the aggregative response by the natural enemies to different host densities, achieved through host attractance and arrestment. All exotic natural enemies, except the males ofApoanagyrus spp., were fast attracted to the host colonies. As compared to the control tips, they concentrated on the infested tips about 50-fold for the twoApoanagyrus spp. and 10 to 20-fold for the exotic coccinellids. By contrast, non-coevolved indigenous coccinellids, as well as generalist predators like ants and spiders were attracted to the infested tips only 2 to 5-fold.A. lopezi responded best to different host densities, followed byA. diversicornis and the coccinellids, followed by ants and spiders. None of the parasitoids or predators was particularly attracted to egg masses. These results correspond closely to the known efficiencies of these natural enemies,A. lopezi standing out among all candidates. The results of such aggregation studies are compared with those of life-table studies.     

Spontane und induzierte Metamorphose bei Larven des Krallenfrosches (Xenopus laevis Daud.) im Kälteversuch

Zusammenfassung Bei jungenXenopuslarven (Prämetamorphose) ist die Mortalität im Bereich von 8,5–22° C konstant, nimmt jedoch unterhalb von 8,5° C stark zu.Die Wirkung der Kälte (10° C) auf die spontane Metamorphose zeigt stadienspezifische Unterschiede. Eine vollkommene Metamorphosehemmung gelingt nur bei Prämetamorphoselarven; bei älteren Larven (Prometamorphose) ist die Umwandlung nur verzögert.Blockierte Larven können in der Kälte durch Behandlung mit Thyroxin bzw. TSH zur Metamorphose veranlaßt werden. Da weder die Ansprechbarkeit larvaler Gewebe auf Thyroxin, noch diejenige der Schilddrüse auf TSH betroffen sind, muß die kältebedingte Metamorphosehemmung auf einer Blockierung des übergeordneten Steuerungszentrums beruhen.

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Spontaneous and induced metamorphosis inXenopus larvae at low temperature

Summary In premetamorphicXenopus larvae mortality is not influenced by lowering the temperature from 22° to 8.5° C, but it rises dramatically below 8.5° C. At 10° C complete inhibition of spontaneous metamorphosis occurs only in premetamorphic larvae; beyond stage 56 cold treatment only delays the metamorphic changes.In cold-arrested premetamorphic larvae thyroxine and TSH elicit metamorphic responses. Since the responding capacity of the larval tissues to thyroxine is not affected, blockage of spontaneous metamorphosis at low temperature must result from the inhibition of the hypothalamic center.

     

Increased frequency of dividing microspores and improved maintenance of multicellular microspores of Coffea arabica in medium with coconut milk

The number of dividing microspores of Coffea arabica L. cv. Catuai and Catimor could be drastically increased in microspore media containing 16% (w/v) coconut milk, allowing cell divisions to continue in the microspore and multicellular microspores to survive until day 60. After a cold treatment, the microspores were mechanically isolated prior to cultivation in Murashige and Skoog medium supplemented with sucrose and maltose and (mg l^-1) 2,4-d: 2, BAP: 1 or a combination of kinetin: .5, 2,4-d: .5 and NAA: .5 as stationary suspension at a density of 1,200/ml. The crucial stage during microsporogenesis suitable for in vitro androgenesis proved to be mid uninucleate till early binucleate in flowerbuds with the size of 13–15 mm two to three days before anthesis. The initial steps of androgenesis were determined.Abbreviations BAP 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxy-acetic acid - NAA 1-naphthaleneacetic acid     

THE EVOLUTION OF XY RECOMBINATION: SEXUALLY ANTAGONISTIC SELECTION VERSUS DELETERIOUS MUTATION LOAD

Recombination arrest between X and Y chromosomes, driven by sexually antagonistic genes, is expected to induce their progressive differentiation. However, in contrast to birds and mammals (which display the predicted pattern), most cold‐blooded vertebrates have homomorphic sex chromosomes. Two main hypotheses have been proposed to account for this, namely high turnover rates of sex‐determining systems and occasional XY recombination. Using individual‐based simulations, we formalize the evolution of XY recombination (here mediated by sex reversal; the “fountain‐of‐youth” model) under the contrasting forces of sexually antagonistic selection and deleterious mutations. The shift between the domains of elimination and accumulation occurs at much lower selection coefficients for the Y than for the X. In the absence of dosage compensation, mildly deleterious mutations accumulating on the Y depress male fitness, thereby providing incentives for XY recombination. Under our settings, this occurs via “demasculinization” of the Y, allowing recombination in XY (sex‐reversed) females. As we also show, this generates a conflict with the X, which coevolves to oppose sex reversal. The resulting rare events of XY sex reversal are enough to purge the Y from its load of deleterious mutations. Our results support the “fountain of youth” as a plausible mechanism to account for the maintenance of sex‐chromosome homomorphy.     

The intrinsic ATPase activity of Mycobacterium tuberculosis DnaA promotes rapid oligomerization of DnaA on oriC

Oligomerization of the initiator protein, DnaA, on the origin of replication (oriC) is crucial for initiation of DNA replication. Studies in Escherichia coli (Gram-negative) have revealed that binding of DnaA to ATP, but not hydrolysis of ATP, is sufficient to promote DnaA binding, oligomerization and DNA strand separation. To begin understanding the initial events involved in the initiation of DNA replication in Mycobacterium tuberculosis (Gram-positive), we investigated interactions of M. tuberculosis DnaA (DnaA(TB)) with oriC using surface plasmon resonance in the presence of ATP and ADP. We provide evidence that, in contrast to what is observed in E. coli, ATPase activity of DnaA(TB) promoted rapid oligomerization on oriC. In support, we found that a recombinant mutant DnaA(TB) proficient in binding to ATP, but deficient in ATPase activity, did not oligomerize as rapidly. The corresponding mutation in the dnaA gene of M. tuberculosis resulted in non-viability, presumably due to a defect in oriC-DnaA interactions. Dimethy sulphate (DMS) footprinting experiments revealed that DnaA(TB) bound to DnaA boxes similarly with ATP or ADP. DnaA(TB) binding to individual DnaA boxes revealed that rapid oligomerization on oriC is triggered only after the initial interaction of DnaA with individual DnaA boxes. We propose that ATPase activity enables the DnaA protomers on oriC to rapidly form oligomeric complexes competent for replication initiation.     

Picogram cloning and direct in situ sequencing of DNA from gel pieces.

We describe a simple and rapid procedure for cloning and sequencing of DNA fragments separated by gel electrophoresis, using novel hydrophilic gels, Clearose BG, Spreadex, and Poly(NAT), that do not melt at 95 degrees C. For cloning, a band of interest is excised precisely and incubated in an extraction buffer containing 5-10 mM MgCl2 at 70 degrees C for 15-45 min. The eluted DNA is added directly to the plasmid solution. Using a topoisomerase-based ligation system, we were able to transform bacteria with a few picograms of DNA and isolate recombinant clones. For in situ sequencing, the DNA in the gel serves as the template. No treatment before cycle sequencing is necessary for fragments up to 500 bp.