Development of polymorphic markers for Cirsium arvense,Canada thistle,and their amplification in closely related taxa

Suppression of invasive Canada thistle, Cirsium arvense, with biological control agents has stalled because introduced agents were not host‐specific. To aid in the development of more effective management strategies, molecular markers are needed to examine the genetic structure of Canada thistle populations. Microsatellite (simple sequence repeat) markers were developed and intersimple sequence repeat (ISSR) markers were tested for North American populations. An average of nine polymorphic alleles per microsatellite locus and 11 per ISSR locus were detected. These will be used to examine the genetic structure of C. arvense in the northern Great Plains and their transferability to endemic Cirsium spp.     

Host finding and recognition by Altica carduorum, a defoliator of Cirsium arvense

Altica carduorum Guer. (Coleoptera: Chrysomelidae) populations aggregate on vigorously growing stands of its host, the thistle Cirsium arvense (L.) Scop. (Asteraceae). The beetle needs contact to recognize a Cirsium species and biting to distinguish its host C. arvense. Feeding or mechanical damage on C. arvense, but not other Cirsium species, aggregates the beetles. Also feces from feeding on C. arvense, but not other Cirsium species, aggregates the opposite sex. Adults also aggregate on larval feces. We suggest that initial host location in the spring depends on random encounter by beetles overwintering in a stand of C. arvense. The beetles commonly make short flights after feeding and either land on another C. arvense plant, which establishes a secondary aggregation centre, or return to the original one. In this manner beetles released on a stand of C. arvense in China spread a radius of 600 m in a year, but those released 35 m away from their host failed to find it. Altica carduorum will develop on any Cirsium species to which it is confined, but is monophagous in the field because host finding is dependent on aggregation to wound and feces substances that are specific to C. arvense.     

Infestation of a novel host plant by <Emphasis Type="Italic">Tephritis conura</Emphasis> (Diptera: Tephritidae) in northern Britain: host-range expansion or host shift?

The addition of a novel host plant to a phytophagous insect’s diet may result in subsequent host-plant specialisation, and is believed to be a key cause for speciation in this trophic group. In northern Britain, the tephritid fly Tephritis conura has experienced a unique host-plant expansion, from the melancholy thistle Cirsium heterophyllum to the marsh thistle C. palustre. Here, we examine whether the incorporation of C. palustre in the repertoire of British T. conura flies has caused genetic divergence between populations infesting the old host and the novel host, and how British populations differ from populations infesting C. heterophyllum in continental Europe where C. palustre is not infested. No evidence for restricted gene flow among British C. palustre and C. heterophyllum flies was found. Significant differentiation between British and continental T. conura was found at only one allozyme locus, hexokinase, and caused by a new allele, Hex_95. Hexokinase is related to host-race formation in continental European flies infesting C. heterophyllum and C. oleraceum, and might be linked to loci determining host choice. Based on morphological and phenological data from previous studies, we suggest that T. conura in Britain has adapted to the novel host but that host-race formation is impeded by similar plant phenologies.     

Microsatellite markers for evolutionary studies in the sexually deceptive orchid genus Chiloglottis

The orchids in the genus Chiloglottis are pollinated exclusively by sexual deception. We describe 20 variable microsatellite loci that will facilitate the study of the evolution and ecology of this specialized pollination system. Two loci were isolated from Chiloglottis trilabra, with the rest derived from Chiloglottis valida. All loci were polymorphic in C. valida (H_E 0.07–0.87). Most loci amplified, and were variable within close relatives of C. valida, and a number amplified more broadly, including outside the genus.     

Microsatellite markers for <Emphasis Type="Italic">Pseudopanax</Emphasis> trees and their cross-species amplification in the Araliaceae

In order to study hybridisation, taxonomic boundaries and phylogeography in the genus Pseudopanax (Araliaceae), we developed seven novel microsatellite loci from enriched genomic libraries of P. lessonii and P. crassifolius. These loci were characterised in 16 individuals from a single population of P. crassifolius, and displayed 2–11 alleles per locus. Observed heterozygosities ranged from 0.063 to 0.688. Most loci were polymorphic in the closely related Pseudopanax species, and several loci amplified widely across the Araliaceae.     

Isolation and characterization of microsatellite loci in the marine gastropod Nucella lapillus

Our paper deals the cloning and characterization of microsatellites from Nucella lapillus, and tests cross‐species amplification in a congener and in two species of the confamilial genus Thais. Fourteen of 31 microsatellite loci tested were polymorphic, with 4–9 (mean 5.93) alleles per locus. The observed heterozygosity per locus varied from 0.10 to 0.85 (mean 0.37) and expected heterozygosity from 0.48 to 0.85 (mean 0.65). Most primer pairs were successfully amplified in N. freycineti, although only one primer pair was successfully amplified in both species of Thais. The markers are potentially useful for other species of Nucella.     

Effects of pasture competition and specialist herbivory on the performance of Cirsium arvense

Combining specialist herbivory with interspecific plant competition can be an effective means of controlling pasture weeds. Cirsium arvense (Canada thistle, Californian thistle, creeping thistle) is one of the worst weeds of pastoral production systems in New Zealand (NZ). The oligophagous leaf-feeding beetle, Cassida rubiginosa, was recently released in NZ for control of C. arvense. To assess the impact of this biocontrol agent we conducted an outdoor potted-plant experiment with low and high densities of Cassida larvae combined with different levels of interspecific competition from typical NZ pasture species. Secondly, we carried out a field-release experiment to quantify the impact of high densities of Cassida under more natural conditions. Interspecific competition reduced all measured plant parameters of C. arvense except mean shoot height and base diameter. Herbivory by Cassida only reduced root biomass, and showed a weak additive response when combined with competition. All other measured parameters of C. arvense showed a substitutive response, with competition being the only factor having a significant impact on the weed. There were no significant synergistic interactions with competition and herbivory on C. arvense. Interestingly, the number of root buds per plant was significantly greater in the presence of herbivory by Cassida, suggesting that C. arvense may compensate for defoliation. Similar to the potted-plant experiment, Cassida had no significant effect on shoot growth and development in the field-release experiment. The results of this study indicate that competition from typical NZ pasture species is a more important factor than herbivory by Cassida, and unless Cassida reaches outbreak densities, it will likely have an insignificant impact on this weed.     

Eighteen microsatellite loci for endemic African weakly electric fish (Campylomormyrus,Mormyridae) and their cross species applicability among related taxa

We describe isolation and characterization of the first microsatellite loci specifically developed for African weakly electric fish (Mormyridae), for the genus Campylomormyrus. Seventeen of our 18 loci are polymorphic within the Campylomormyrus numenius species complex. The polymorphic loci showed four to 15 alleles per locus, an expected heterozygosity between 0.46 and 0.94, and an observed heterozygosity between 0.31 and 1.00. Most primers also yield reproducible results in several other mormyrid species. These loci comprise a set of molecular markers for various applications, from moderately polymorphic loci suitable for population studies to highly polymorphic loci for pedigree analysis in mormyrids.     

Development of SSR markers for typing cultivars in the mushroom Auricularia auricula-judae

Auricularia auricula-judae, also known as black wood ear, is one of the most popular edible mushrooms in China. But the confusion about cultivars has limited the development of A. auricula-judae production. In this article, 17 polymorphic SSR markers were cloned and used to differentiate the cultivars of A. auricula-judae. The polymorphism information content (PIC) of these SSR ranged from 0.10 to 0.84, while the average was 0.47. The number of alleles detected for each locus was 2–11, with an average of 4.7 alleles per locus. The dendrogram, based on 17 SSR markers by UPGMA clustering, could differentiate the 16 A. auricula-judae cultivars in this study. In fact, the 16 cultivars analyzed in this study could be efficiently differentiated using a combination of three polymorphic SSR loci with high PIC. The total of 17 polymorphic SSR loci could also be amplified correctly in the A. polytricha strains surveyed. This is the first report on the development of SSR markers in the genus Auricularia.     

Microsatellite markers for two species of dampwood termites in the genus Zootermopsis (Isoptera: Termopsidae)

Dampwood termites in the genus Zootermopsis inhabit forested areas in western North America. To better understand the colony composition and breeding structure of Zootermopsis, we identified polymorphic microsatellite loci to use in population analysis. Microsatellite loci were isolated from Zootermopsis nevadensis nevadensis (Hagen); however, all primers amplified homologous loci in Zootermopsis angusticollis (Hagen) and Zootermopsis nevadensis nuttingi (Hagen). Twelve loci were polymorphic in one or more of the above subspecies and species. The number of alleles per locus ranged from one to six, with some allelic differences among subspecies and species. We are currently utilizing the microsatellite markers to investigate the population genetics of Zootermopsis.     

Characterization of microsatellite loci in village indigobirds Vidua chalybeata and cross‐species amplification in estrildid and ploceid finches

We characterized 11 microsatellite primer pairs for the village indigobird Vidua chalybeata. The loci were highly polymorphic, with 7–13 alleles per locus. Gene diversity, estimated as expected heterozygosity, ranged from 0.52 to 0.86, and was generally matched by levels of observed heterozygosity (0.49–0.91). Many of these primer pairs amplified polymorphic loci in cross‐species amplification trials with a variety of estrildid and ploceid finches and a sparrow, Passer griseus. These primers will be valuable for genetic analyses of the brood parasitic indigobirds and whydahs (genus Vidua) as well as other Old World finches.     

Isolation and characterization of polymorphic microsatellite loci from black sea bass (Centropristis striata) and cross-species amplification

Black sea bass (Centropristis striata) is an economically important serranid species. A number of 39 microsatellite loci were isolated from two enriched genomic library of C. striata. Eleven of these loci were polymorphic in a test population with alleles per locus ranging from 3 to 8, and observed and expected heterozygosities per locus from 0.26 to 1.00 and from 0.61 to 0.84, respectively. Four loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. Cross-species amplification of these polymorphic microsatellite loci was performed in additional two related species. These polymorphic microsatellite loci would be useful for investigating genetic resource of C. striata and other related species.     

Development of microsatellite loci for Pelophylax plancyi and cross-amplification in other ranid species

Thirteen polymorphic microsatellite loci were obtained from the gold-striped pond frog, Pelophylax plancyi. The number of alleles per locus ranged from 5 to 13 with an average of 8.38. The observed and expected heterozygosities (H _O and H _E) ranged from 0.533 to 0.890 and from 0.620 to 0.897, with averages of 0.686 and 0.751, respectively. In order to assess interspecific amplification, all primer pairs were tested with the same PCR conditions on three other Pelophylax species: Pelophylax hubeiensis, Pelophylax fukienensis, Pelophylax nigromaculata and two species from the genus Hylarana and Hoplobatrachus, respectively. All of these loci can be amplified successfully in the Pelophylax species, with eight loci amplified in all species tested.     

Isolation and characterization of polymorphic microsatellite loci in Acanthoscelides obtectus Say (Coleoptera: Bruchidae)

Six microsatellite loci were isolated from the bruchid Acanthoscelides obtectus Say (Coleoptera: Bruchidae). Each locus was polymorphic, with the number of alleles ranging from 3 to 18. We found high levels of within‐population variation at most loci, with heterozygosities ranging from 0 to 0.75. Cross‐species amplification of these loci was tested in two other species of the genus Acanthoscelides, A. obvelatus Bridwell and A. argillaceus Sharp.     

Polymorphic microsatellite loci in Plantago lanceolata

The genus Plantago is particularly interesting for evolutionary studies because of its wide range of mating systems. We have developed primers for five highly polymorphic microsatellite loci isolated from P. lanceolata. All five loci amplified and were polymorphic in the two populations examined, Lowsteads Beach in the United Kingdom and Duke in the United States. These new markers will allow a comparison of population structure between the outcrossing species P. lanceolata, and the highly selfing species P. major.     

Mowing during rainfall enhances the control of Cirsium arvense

Pastoral farmers in New Zealand have described dramatic demises in populations of the weed Cirsium arvense (L.) Scop., a perennial herb indigenous to Eurasia, following its mowing during rainfall. To test the hypotheses (1) that the mowing of C. arvense during rainfall increases the control of this weed and (2) that the causal organism in this ‘mowing-in-the-rain’ effect is the vascular wilt fungus Verticillium dahliae, two series of field experiments were carried out in C. arvense-infested pastures in New Zealand, one in autumn 2008 (Experiment series 1, 9 farms), and another in spring/summer 2008–2009 (Experiment series 2, 12 farms). The effect of mowing in the rain as compared to mowing in the dry was to reduce the % ground cover of the thistle in the spring following treatment by 21 and 32% in Experiment series 1 and 2, respectively. Correlations of this ‘rain versus dry’ effect with the incidence of V. dahliae in the subterranean parts of C. arvense shoots sampled in each of the two field experiment series provided no statistical evidence that the effect increased with V. dahliae incidence. Thus these experiments provide no support for the hypothesis that V. dahliae is the biological mechanism for the ‘mowing-in-the-rain’ effect. Nevertheless, they do support mowing during rainfall as a simple and effective management tactic for C. arvense.     

Isolation and characterization of polymorphic microsatellite loci in Acanthoscelides obvelatus Bridwell (Coleoptera: Bruchidae)

Five microsatellite loci were isolated from the bruchid Acanthoscelides obvelatus Bridwell (Coleoptera: Bruchidae). Each locus was polymorphic, with the number of alleles ranging from two to 15. We found high levels of within‐population variation at most loci, with heterozygosity ranging from 0.182 to 0.900. Cross‐species amplification of these loci was tested in two other species of the genus Acanthoscelides, A. obtectus Say and A. argillaceus Sharp.     

Polymorphic microsatellite markers for the Iberian rock lizard species,Iberolacerta cyreni,and cross‐species priming in other Iberolacerta species

Eight polymorphic microsatellite loci are described for the Iberian rock lizard species, Iberolacerta cyreni. Loci were isolated from a partial genomic library that had been enriched for AAAG repeat sequence. Number of alleles per locus ranged from two to 13 in a sample of 24 individuals from the Sierra de Guadarrama (central Spain). Observed heterozygosity ranged from 0.13 to 0.96. At least three loci were amplified and polymorphic in four other Iberian rock lizard species: Iberolacerta monticola, Iberolacerta bonnali, Iberolacerta aranica and Iberolacerta aurelioi. These markers will be used to study mating strategies and reproductive success in I. cyreni.     

Characterization of microsatellite loci in White‐chinned Petrel (Procellaria aequinoctialis) and cross‐amplification in six other procellariiform species

Six polymorphic dinucleotide microsatellite loci were isolated and characterized from the White‐chinned Petrel Procellaria aequinoctialis, using a degenerate primer and PCR‐based technique to construct and screen an enriched genomic library. Preliminary data on three populations show heterozygosity levels ranging from 0.22 to 0.67 and allele numbers from three to nine. Preliminary data also suggest genetic distance between these three populations (F_ST 0.088). Cross‐species amplification of these six microsatellite loci and one further locus were tested in six other procellariiform species of the genus Procellaria, Macronectes, Thalassarche and Diomedea.     

Localization of 5S rRNA Loci in Three Coregonid Species (Salmonidae)

In the present study the chromosome distribution of the 5S rDNA loci and its relation to the major rDNA genes were investigated in three Coregonid species (Salmonidae): Coregonus lavaretus, Coregonus peled and Coregonus albula, a family which has experienced large karyotype rearrangements along its evolution starting from a tetraploid ancestor. 5S PRINS/CMA₃ sequential staining together with previous data enabled us to locate 5S rRNA genes and nucleolar organizer regions (NORs) in the three species analyzed. PRINS revealed the 5S rDNA cluster at the distal part of the long arm of a similar submetacentric chromosome pair in the three species. Our data indicate that 5S rDNA clusters have probably conserved chromosomal location in the genus Coregonus, whereas 45S rDNA (NOR) sites are clearly differentiated, from a single locus in C. peled, to multiple loci in C. lavaretus and highly polymorphic multichromosomal location in C. albula.