Nine polymorphic microsatellite markers in Centaurea stoebe L. [subspecies C. s. stoebe and C. s. micranthos (S. G. Gmelin ex Gugler) Hayek] and C. diffusa Lam. (Asteraceae)

Centaurea stoebe (subspecies C. s. stoebe and C. s. micranthos[S. G. Gmelin ex Gugler) Hayek] and Centaurea diffusa are Eurasian plant species that have invaded much of North America. We isolated seven microsatellite loci from C. stoebe and two loci from C. diffusa. All loci described here amplify in both species and have between six and 25 alleles each. These markers will be useful in examining population structure and addressing questions regarding these invasions.     

Revised methods for the mass-rearing of the spotted knapweed biological control agent,Cyphocleonus achates (Coleoptera: Curculionidae), in field corrals

Cyphocleonus achates (Fahraeus), a root-feeding weevil introduced from Eurasia, is an effective biological control agent against spotted knapweed, Centaurea stoebe L. ssp. micranthos. Because C. achates is univoltine and does not fly, distribution of the weevil has been slow. To hasten the weevil's distribution, a rearing effort using field corrals was initiated at a facility in Corvallis, Montana. Procedures for mass-rearing the weevil in field corrals are described, with an emphasis on improvements over earlier methods. The described field-corral approach is effective and appropriate for producing C. achates for distribution in the western United States.     

The lesser of two weevils: physiological responses of spotted knapweed (Centaurea stoebe) to above- and belowground herbivory by Larinus minutus and Cyphocleonus achates

The physiological responses of plants to variable levels of root and shoot herbivory in the field may yield valuable insights regarding potential compensation or tolerance responses for herbivory. In an infestation of Centaurea stoebe (spotted knapweed) located in the Colorado foothills, we measured physiology, biomass, and flower production of individual plants undergoing a natural range of herbivory by the above- and belowground biological control insects, Larinus minutus and Cyphocleonus achates. Over the growing season, net carbon assimilation rate, transpiration, stomatal conductance, and intercellular leaf [CO₂] (C _i) all decreased, while water use efficiency increased. The decrease in these physiological traits was due to an increase in the intensity of L. minutus damage over time; effects of C. achates root damage to plant physiology, including transpiration were only marginally significant. The effects of these two species on plant physiology were not interactive, and Larinus minutus was found to exert larger negative effects on this invasive plant in terms of plant physiology and potential reproductive output than C. achates. While previous studies have shown C. achates to have significant negative effects on population densities of spotted knapweed, the addition of Larinus minutus to the suite of insects used in biological control of spotted knapweed should facilitate continued or enhanced reduction in densities of this noxious weed.     

Interspecific interactions between the gall-fly Urophora affinis Frfld. (Diptera: Tephritidae) and the weevil Larinus minutus Gyll. (Coleoptera: Curculionidae), two biological control agents released against spotted knapweed, Centaurea stobe L. ssp. micranthos

Interspecific competition has been suggested as an explanation for the failure of some insects as biological control agents for weeds. Enclosure and exclusion cages were used, in southern British Columbia, Canada to evaluate the importance of interspecific competition between a seedhead weevil, Larinus minutus, and a gall-inducing fly, Urophora affinis, two biocontrol agents released against spotted knapweed in North America. At the seedhead scale, U. affinis, which is an inferior biological control agent based on knapweed seed mortality, was the superior competitor. Larinus minutus attack rates were significantly lower in the presence of U. affinis compared to release treatments where L. minutus was attacking alone. Reduced L. minutus attack rates were apparent in seed heads expected to contain both species, assuming insect distributions were random, but instead only contained U. affinis. L. minutus did not significantly affect U. affinis density. Although overall attack rates on knapweed seedheads were higher when both species were together at a site, the consequence of the antagonistic interaction is that overall seed destruction was not as high as it could have been if the weevil were attacking on its own. These results support minimizing the number of biocontrol agents released that use similar resources on the target weed, to avoid negative interactions between control agents and potential reductions in biocontrol efficacy resulting from competitive exclusion.     

Additive effects of aboveground and belowground herbivores on the dominance of spotted knapweed (Centaurea stoebe)

Spotted knapweed (Centaurea stoebe) is found in over 3 million ha of rangeland and forests across North America, and evidence supporting the use of biological control as a regional method to reduce infestations and their associated impacts remains inconclusive. Several species of insects have been reported to reduce plant densities in some areas; however, rigorous studies that test combinations of these species and the influence of resource availability are lacking. We examined the singular and combined effects of herbivory by a root weevil (Cyphocleonus achates) and a flower head weevil (Larinus minutus) on the growth and flower production of C. stoebe. We also manipulated soil resource fertility as an additional factor that could explain the outcomes of contradictory biological control herbivore effects on C. stoebe. In a greenhouse study, herbivory by C. achates decreased flower production for plants across all resource environments. In a caged common garden study, the negative effects of herbivory also did not interact with soil nutrient status. However, the presence of plant competition further decreased knapweed growth, and the negative effects of concurrent herbivory by C. achates and L. minutus on plant biomass and flower production were additive. Derived within the context of variable levels of soil nutrient availability and competing vegetation, these results support the cumulative stress hypothesis and the contention that combined above- and belowground herbivory can reduce spotted knapweed densities and reduce the ecological and economic impacts of this species in rangelands of western North America.     

Genetic patterns across multiple introductions of the globally invasive crab genus Carcinus

The European green crab Carcinus maenas is one of the world's most successful aquatic invaders, having established populations on every continent with temperate shores. Here we describe patterns of genetic diversity across both the native and introduced ranges of C. maenas and its sister species, C. aestuarii, including all known non‐native populations. The global data set includes sequences from the mitochondrial cytochrome c oxidase subunit I gene, as well as multilocus genotype data from nine polymorphic nuclear microsatellite loci. Combined phylogeographic and population genetic analyses clarify the global colonization history of C. maenas, providing evidence of multiple invasions to Atlantic North America and South Africa, secondary invasions to the northeastern Pacific, Tasmania, and Argentina, and a strong likelihood of C. maenas × C. aestuarii hybrids in South Africa and Japan. Successful C. maenas invasions vary broadly in the degree to which they retain genetic diversity, although populations with the least variation typically derive from secondary invasions or from introductions that occurred more than 100 years ago.     

Influence of Centaurea biebersteinii patch size on Urophora quadrifasciata (Dipt. Tephritidae) in Michigan, USA

Abstract:  Host plant patch size plays an important role in the distribution and densities of many insect species. Understanding how the host plant patch size influences the population of a biological control agent is necessary to monitor the success of a biological control programme. Urophora quadrifasciata (Dipt. Tephritidae) was released in the early 1970s to control the spotted knapweed, Centaurea biebersteinii (Asteraceae) in North America. The studies reported here investigated the influences of spotted knapweed patch size and stem density on U. quadrifasciata larval populations overwintering within capitula in the Upper Peninsula of Michigan, USA. An inverse relationship was found between the percentage of capitula with U. quadrifasciata larvae and spotted knapweed patch size in 2002 from Houghton county, MI. In 2003, an inverse relationship was also found in the south region of the study area in Houghton and Keweenaw counties, Michigan. The percentage of capitula with U. quadrifasciata larvae and spotted knapweed patch size were positively related for the central region. No relationship was found for the North region in 2003. Dispersal of U. quadrifasciata is directed north-east along the Keweenaw Peninsula, Michigan. With this directed spread, areas with well-established populations of U. quadrifasciata would provide source individuals for new colonization. Spotted knapweed patch size may play an important role in the overwintering populations of U. quadrifasciata as a result of dispersal.     

Evidence for multiple sources of invasion and intraspecific hybridization in Brachypodium sylvaticum (Hudson) Beauv. in North America

We compared the levels and distribution of genetic diversity in Eurasian and North American populations of Brachypodium sylvaticum (Huds.) Beauv. (false brome), a newly invasive perennial bunchgrass in western North America. Our goals were to identify source regions for invasive populations, determine the number of independent invasion events, and assess the possibility that postinvasion bottlenecks and hybridization have affected patterns of genetic diversity in the invaded range. We tested the hypothesis that this Eurasian grass was accidentally introduced into two areas in Oregon and one site in California by examining nuclear microsatellites and chloroplast haplotype variation in 23 introduced and 25 native populations. In the invaded range, there was significantly lower allelic richness (R(S)), observed heterozygosity (H(O)) and within-population gene diversity (H(S)), although a formal test failed to detect a significant genetic bottleneck. Most of the genetic variation existed among populations in the native range but within populations in the invaded range. All of the allelic variation in the invaded range could be explained based on alleles found in western European populations. The distribution of identified genetic clusters in the North American populations and the unique alleles associated with them is consistent with two historical introductions in Oregon and a separate introduction to California. Further analyses of population structure indicate that intraspecific hybridization among genotypes from geographically distinct regions of western Europe occurred following colonization in Oregon. The California populations, however, are more likely to be derived from one or perhaps several genetically similar regions in the native range. The emergence and spread of novel recombinant genotypes may be facilitating the rapid spread of this invasive species in Oregon.     

Evidence for multiple introductions of Phragmites australis to North America: detection of a new non-native haplotype

We found a new non-native haplotype of Phragmites australis in North America that provides convincing evidence for multiple introductions of this highly invasive reed from Europe. Prior to our detection of this new non-native haplotype, invasion of North America by this reed grass was thought to be limited to a single cp-DNA haplotype–haplotype M. However, we found two sites colonized by haplotype L1 in Quebec, Canada, a haplotype native to northern Europe, Great Britain and Romania. Because the invasion of North America by P. australis is ongoing, and because there is evidence for intra- and inter-specific hybridization and increased fecundity resulting from outcrossing, more attention should be paid to genetic differences and associated vigor of populations of introduced Phragmites across North America.     

Reduced genetic diversity, increased isolation and multiple introductions of invasive giant hogweed in the western Swiss Alps

The giant hogweed ( Heracleum mantegazzianum ) has successfully invaded 19 European countries as well as parts of North America. It has become a problematic species due to its ability to displace native flora and to cause public health hazards. Applying population genetics to species invasion can help reconstruct invasion history and may promote more efficient management practice. We thus analysed levels of genetic variation and population genetic structure of H. mantegazzianum in an invaded area of the western Swiss Alps as well as in its native range (the Caucasus), using eight nuclear microsatellite loci together with plastid DNA markers and sequences. On both nuclear and plastid genomes, native populations exhibited significantly higher levels of genetic diversity compared to invasive populations, confirming an important founder event during the invasion process. Invasive populations were also significantly more differentiated than native populations. Bayesian clustering analysis identified five clusters in the native range that corresponded to geographically and ecologically separated groups. In the invaded range, 10 clusters occurred. Unlike native populations, invasive clusters were characterized by a mosaic pattern in the landscape, possibly caused by anthropogenic dispersal of the species via roads and direct collection for ornamental purposes. Lastly, our analyses revealed four main divergent groups in the western Swiss Alps, likely as a consequence of multiple independent establishments of H. mantegazzianum .     

Strategies for ex situ conservation of Centaurea cineraria subsp. circae (Asteraceae), an endemic plant from Lazio (Italy)

Centaurea cineraria subsp. circae is an endemic plant with a distribution area limited to Circeo mountain (Lazio, Italy), whose population was estimated in a very low number of individuals. The aim of this work was to investigate ex situ conservation strategies such as achene collection and in vitro plant propagation, which will permit to carry out restoration programmes. The test carried out on the achenes demonstrated that only 5.5% of them were morphologically healthy. Seed germination tests showed that seeds do not display dormancy and that germination does not require pre-treatments. The higher germination rate (67.5%) was observed under a photoperiod of 12/12 h (light/dark) and temperature regime +20/+10°C. The in vitro studies demonstrated that micropropagation, acclimatization and the transfer outdoors of C. cineraria subsp. circae are not particularly difficult: 74% of shoot explants in a Murashige and Skoog (MS) medium added with 0.5 mg/l benzylaminopurine and 2 mg/l kinetin formed multiple shoots; 100% of shoots rooted in the MS medium added with 0.5 mg/l indole-3-butyric acid and over 90% survived the acclimatization phase. After been transferred outdoors, the totality of in vitro-propagated plants bloomed and appeared morphologically indistinguishable from wild plants. Preliminary chemical analyses showed a similar profile for in vitro-propagated and wild plants.     

Effects of the Invasive Forb <Emphasis Type="Italic">Centaurea maculosa</Emphasis> on grassland Carbon and Nitrogen Pools in Montana,USA

Invasions by exotic forbs are changing large areas of North American grasslands, but their biogeochemical impacts are not well characterized. Additionally, although many invasive plants may alter biogeochemistry, an invasive species effects have rarely been evaluated across physically diverse sites. We sampled nine sites containing the perennial Eurasian forb Centaurea maculosa to determine if this invasive species alters soil C and N pools in native grasslands in Montana, USA. We sampled surface soil in adjacent microsites with C. maculosa and native grasses and analyzed soil C and N pools with slow to rapid turnover. None of the pools evaluated in the laboratory showed significant differences between C. maculosa and grass microsites when analyzed across all sites. Some differences were found at individual sites, but they were infrequent and inconsistent: Four sites had no differences, four had differences in one or two pools with intermediate (particulate organic matter C or N) or rapid turnover rates (potentially mineralizable N), and just one site had differences encompassing pools with rapid, intermediate, and slow (total C and N, silt-and-clay-associated N) turnover rates. Where they differed, pools were usually smaller under C. maculosa plants than under native grasses, but the opposite was found at one site. In situ N availability, estimated using ion exchange resins, was significantly lower under C. maculosa than under grasses at one of three sites sampled. Results indicate that C. maculosa may sometimes reduce soil C and N pools, including those related to N availability, but they argue against generalizing about the impacts of C. maculosa in grasslands.     

Isolation and characterization of polymorphic microsatellites in the Potato Tuber Moth Tecia solanivora (Povolny, 1973) (Lepidoptera: Gelechiidae)

Nine polymorphic microsatellite markers were isolated from Tecia solanivora, one of the most serious pests of potato tubers in Central and South America. As found in other studies of Lepidoptera, development of microsatellites is a difficult task: in our case, despite the large number of clones sequenced (796), of which 70 were unique, only nine loci were found to be both variable, and in Hardy-Weinberg equilibrium, No null alleles were detected. The loci were tested in three other co-occurring Gelechiidae species, one of which was variable. These loci will be used to provide a greater understanding of the genetic changes occurring during the invasive process in this species.     

Isolation and characterization of polymorphic microsatellite markers from fireweed,Senecio madagascariensis Poir. (Asteraceae)

Ten polymorphic microsatellite loci were isolated and characterized from invasive fireweed (Senecio madagascariensis) populations in the Hawaiian archipelago. These loci provided markers with polymorphism of six to 24 alleles per locus within 96 individuals collected from two populations from the island of Maui. The expected and observed heterozygosities ranged from 0.31 to 0.91 and from 0.056 to 1. These markers should be useful to study the importance of genetic diversity in invasion success of this species.