Feral populations of Brassica oleracea along Atlantic coasts in western Europe.

There has been growing emphasis on the role that crop wild relatives might play in supporting highly selected agriculturally valuable species in the face of climate change. In species that were domesticated many thousands of years ago, distinguishing wild populations from escaped feral forms can be challenging, but reintroducing variation from either source could supplement current cultivated forms. For economically important cabbages (Brassicaceae: Brassica oleracea), "wild" populations occur throughout Europe but little is known about their genetic variation or potential as resources for breeding more resilient crop varieties. The main aim of this study was to characterize the population structure of geographically isolated wild cabbage populations along the coasts of the UK and Spain, including the Atlantic range edges. Double-digest restriction-site-associated DNA sequencing was used to sample individual cabbage genomes, assess the similarity of plants from 20 populations, and explore environment-genotype associations across varying climatic conditions. Interestingly, there were no indications of isolation by distance; several geographically close populations were genetically more distinct from each other than to distant populations. Furthermore, several distant populations shared genetic ancestry, which could indicate that they were established by escapees of similar source cultivars. However, there were signals of local adaptation to different environments, including a possible relationship between genetic diversity and soil pH. Overall, these results highlight wild cabbages in the Atlantic region as an important genetic resource worthy of further research into their relationship with existing crop varieties.

PepA and ArgR do not regulate Cre recombination at the bacteriophage P1 loxP site.

In the lysogenic state, bacteriophage P1 is maintained as a low copy-number circular plasmid. Site-specific recombination at loxP by the phage-encoded Cre protein keeps P1 monomeric, thus helping to ensure stable plasmid inheritance. Two Escherichia coli DNA-binding proteins, PepA and ArgR, were recently reported to be necessary for maintenance or establishment of P1 lysogeny. PepA and ArgR bind to regulatory DNA sequences upstream of the ColE1 cer recombination site to regulate site-specific recombination by the XerCD recombinases. This recombination keeps ColE1 in a monomeric state and helps to ensure stable plasmid maintenance. It has been suggested that ArgR and PepA play a similar role in P1 maintenance, regulating Cre recombination by binding to DNA sequences upstream of loxP. Here, we show that ArgR does not bind to its proposed binding site upstream of loxP, and that Cre recombination at loxP in its natural P1 context is not affected by PepA and ArgR in vitro. When sequences upstream of loxP were mutated to allow ArgR binding, PepA and ArgR still had no effect on Cre recombination. Our results demonstrate that PepA requires specific DNA sequences for binding, and that PepA and ArgR have no direct role in Cre recombination at P1 loxP.

Determinants of product topology in a hybrid Cre-Tn3 resolvase site-specific recombination system.

Many natural DNA site-specific recombination systems achieve directionality and/or selectivity by making recombinants with a specific DNA topology. This property requires that the DNA architecture of the synapse and the mechanism of strand exchange are both under strict control. Previously we reported that Tn3 resolvase-mediated synapsis of the accessory binding sites from the Tn3 recombination site res can impose topological selectivity on Cre/loxP recombination. Here, we show that the topology of these reactions is profoundly affected by subtle changes in the hybrid recombination site les. Reversing the orientation of loxP relative to the res accessory sequence, or adding 4 bp to the DNA between loxP and the accessory sequence, can switch between two-noded and four-noded catenane products. By analysing Holliday junction intermediates, we show that the innate bias in the order of strand exchanges at loxP is maintained despite the changes in topology. We conclude that a specific synaptic structure formed by resolvase and the res accessory sequences permits Cre to align the adjoining loxP sites in several distinct ways, and that resolvase-mediated intertwining of the accessory sequences may be less than has been assumed previously.