Multiple hydroxyphenethyl glucosinolate isomers and their tandem mass spectrometric distinction in a geographically structured polymorphism in the crucifer Barbarea vulgaris.

Two distinct glucosinolate (GSL) chemotypes (P and G-types) of Barbarea vulgaris (Brassicaceae) were known from southern Scandinavia, but whether the types were consistent in a wider geographic area was not known. Populations (26) from Eastern and Central Europe were analyzed for GSLs in order to investigate whether the two types were consistent in this area. Most (21) could be attributed to one of the previously described GSL profile types, the P-type (13 populations) and the G-type (8 populations), based on differences in the stereochemistry of 2-hydroxylation, presence or absence of phenolic glucobarbarin derivatives, and qualitative differences in indole GSL decoration (tested for a subset of 8+6 populations only). The distinction agreed with previous molecular genetic analysis of the same individuals. Geographically, the P-type typically occurred in Eastern Europe while the G-type mainly occurred in Central Europe. Of the remaining five populations, minor deviations were observed in some individuals from two populations genetically assigned to the G-type, and a hybrid population from Finland contained an additional dihydroxyphenethyl GSL isomer attributed to a combinatorial effect of P-type and G-type genes. Major exceptions to the typical GSL profiles were observed in two populations: (1) A G-type population from Slovenia deviated by a high frequency of a known variant in glucobarbarin biosynthesis ('NAS form') co-occurring with usual G-type individuals. (2) A population from Caucasus exhibited a highly deviating GSL profile dominated by p-hydroxyphenethyl GSL that was insignificant in other accessions, as well as two GSLs investigated by NMR, m-hydroxyphenethylGSL and a partially identified m,p disubstituted hydroxy-methoxy derivative of phenethylGSL. Tandem HPLC-MS of seven NMR-identified desulfoGSLs was carried out and interpreted for increased certainty in peak identification and as a tool for partial structure elucidation. The distinct, geographically separated chemotypes and rare variants are discussed in relation to future taxonomic revision and the genetics and ecology of GSLs in B. vulgaris.

Interactive impacts of a herbivore and a pathogen on two resistance types of Barbarea vulgaris (Brassicaceae).

It is well known that pathogens and arthropod herbivores attacking the same host plant may affect each other. Little is known, however, about their combined impact on plant fitness, which may differ from simple additive expectations. In a 2-year common garden field experiment, we tested whether the pathogen Albugo sp. (white blister rust) and the herbivorous flea beetle Phyllotreta nemorum affected each other's performance on two resistance types (G-type and P-type) of the crucifer Barbarea vulgaris ssp. arcuata, and whether biomass, reproduction and survival of the plants were affected by interactive impacts of the antagonists. Most of the insect-resistant G-plants were severely affected by white rust, which reduced biomass and reproductive potential compared to the controls. However, when also exposed to flea beetles, biomass loss was mitigated in G-plants, even though apparent disease symptoms were not reduced. Most of the insect-susceptible P-plants were resistant to white rust; however, the number of flea beetle mines tended to increase in plants also exposed to Albugo, and biomass at the last harvest was slightly lower in the combined treatment. Thus, interactive impacts of the herbivore and pathogen differed between the two resistance types, with an antagonistic combined impact in G-plants, which lasted surprisingly long, and a slight synergistic impact in P-plants.

Different geographical distributions of two chemotypes of Barbarea vulgaris that differ in resistance to insects and a pathogen.

The interactions of plants with herbivores and pathogens have been suggested to drive the evolution of resistances in plants and in some cases new lineages and taxa. However, such divergence may require reproductive isolation, e.g., in allopatry. In the crucifer Barbarea vulgaris, some plants are resistant to the flea beetle Phyllotreta nemorum, due to production of specific saponins, whereas others are susceptible. Resistant and susceptible plants additionally differ in resistance to the pathogen Albugo candida, content of glucosinolates, and leaf pubescence, and they are genetically strongly divergent and partially reproductively incompatible. This suggests that at some point they were separated for a considerable length of time. Previously, the insect susceptible P-type had been described only from Denmark, Sweden, and Estonia, whereas the resistant G-type is widely distributed in Western Europe. Here, we tested whether the two plant types have divergent geographical distributions and maintain their distinct trait associations throughout their range. The insect-susceptible type was found in Russia, the Baltics, and parts of Fennoscandia, but not in Central Europe. In contrast, the insect resistant type was found from Finland and westwards. Their different trait associations were consistent within the two ranges. We therefore suggest that the two plant types diverged in allopatry at some time in the past, and evolved different resistances in response to local antagonists. The two plant types probably maintain their distinctness due to a hybridization barrier. Thus, the present distributions of the two types may be shaped by both historical processes and current differential biotic selection.

Phylogeny of an Albugo sp. infecting Barbarea vulgaris in Denmark and its frequency of symptom development in natural populations of two evolutionary divergent plant types.

The oomycete Albugo candida has long been considered a broad spectrum generalist pathogen, but recent studies suggest that it is diverged into several more specialized species in addition to the generalist Albugo candida sensu stricto. Whereas these species cause the disease white blister rust in many crucifer plants, asymptomatic endophytic infections may be important in the epidemiology of others. One of the plant species attacked by Albugo sp. is the wild crucifer Barbarea vulgaris ssp. arcuata, which is diverged into two phytochemically and genetically different types with different geographical distributions in Europe. These were previously shown to differ strongly in propensity to develop white rust upon controlled infections in the greenhouse. Here, we analyse the phylogenetic relatedness of this local Albugo sp. field isolate to other species and lines of Albugo spp., including others collected on B. vulgaris. We further ask whether the difference in incidence of white rust between the two types of B. vulgaris are also expressed in natural populations. Phylogenetically, the local Albugo sp. field isolate clustered tightly together with previously analysed samples from B. vulgaris, supporting that the Albugo sp. infecting B. vulgaris may indeed be an independent specialized species. White blister rust and Albugo DNA was only detected in two populations of the plant type that frequently develops symptoms upon controlled inoculations. The lack of white rust and Albugo sp. DNA in the other plant type may be due to either resistance, preventing infection, or asymptomatic infection of other tissues than leaves, which we analysed.