Characterization of eight microsatellite loci in Steller sea lions (Eumetopias jubatus)

Steller sea lions (Eumetopias jubatus) are listed as an endangered species in western Alaska and have exhibited a significant population decline throughout their range. Eight microsatellite loci were isolated from genomic DNA libraries. In addition, all these markers were found to be variable in nine individuals of the California sea lion (Zalophus californicus). This panel of markers was developed to analyse population structure in Steller sea lions throughout their range.     

Toxicity of some insecticides to <Emphasis Type="Italic">Tetranychus urticae</Emphasis>, <Emphasis Type="Italic">Neoseiulus californicus</Emphasis> and <Emphasis Type="Italic">Tydeus californicus</Emphasis>

Three mite species are frequently found on vegetable crops in Italy: the pest Tetranychus urticae Koch (Acari: Tetranychidae), the predator Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) and the unspecialised feeder Tydeus californicus (Banks) (Acari: Tydeidae). In laboratory trials, the direct and residual effects of six insecticides recommended for the control of aphids, whiteflies and thrips in vegetable crops, (Biopiren® plus (pyrethrins), Confidor® (imidacloprid), Oikos® (azadirachtin), Plenum® (pymetrozine), Naturalis® (Beauveria bassiana) and Rotena® (rotenone)), were evaluated for the three mite species. All the products affected the mites and their effect was often favourable towards T. urticae and unfavourable towards N. californicus and T. californicus. Rotenone was more toxic to eggs than females of T. urticae. It was highly toxic to N. californicus and caused the death of all treated females of T. californicus. Pyrethrins and imidacloprid increased T. urticae fecundity, but decreased fecundity of N. californicus. Imidacloprid decreased T. californicus fecundity more than pyrethrins. Beauveria bassiana was not toxic to T. urticae and T. californicus, but induced high mortality in the progeny of treated females of N. californicus. Azadirachtin and pymetrozine were the least toxic to T. urticae and N. californicus, but decreased production of larvae in T. californicus. Implications for integrated pest management on vegetables are discussed.     

Characterization of microsatellite DNA markers in a critically endangered species,Mekong giant catfish,Pangasianodon gigas

Microsatellite DNA markers for a critically endangered Mekong giant catfish (Pangasianodon gigas Roberts and Vidthayanon, 1991) were developed from fin clips collected from captive fish using (GT)₁₅ probe. The number of alleles per locus ranged from two to four. The expected heterozygosities ranged from 0.13 to 0.68. Also, these primers were successfully amplified in four closely related species, Pangasius bocourti, Pangasius conchophilus, Pangasius larnaudii and Pangasius sanitwongsei with the number of alleles per locus ranged from 1 to 13, 1 to 16, 1 to 12 and 1 to 4, respectively. These markers should prove to be very useful for the evaluation of genetic diversity for this species and other related Pangasius species.     

Intraguild Predation and Feeding Preferences in Three Species of Phytoseiid Mite Used for Biological Control

The ecological impact of introduced biological control agents on native species of arthropods is a matter of considerable debate. This study investigated the ability of the non-native predatory mite Neoseiulus californicus to feed on the native Typhlodromus pyri and vice versa, as both species now co-occur in UK orchards. Typhlodromips montdorensis is a candidate for introduction into the UK as a glasshouse biological control agent. The ability of T. montdorensis to feed on the widely used N. californicus was investigated to identify possible intraguild predation, which might influence the effectiveness of either or both species as predators of Tetranychus urticae. Both N. californicus and T. pyri consumed larval stages of each other, but in choice experiments both showed a preference for T. urticae. Both N. californicus and T. montdorensis also fed on each other, but whereas N. californicus again showed a preference for T. urticae, T. montdorensis fed equally on T. urticae and N. californicus. Interactions between N. californicus and T. pyri and N. californicus and T. montdorensis are discussed in relation to their effectiveness as biological control agents in the glasshouse and the natural control of spider mite in the field.     

The hidden history of the snowshoe hare,Lepus americanus: extensive mitochondrial DNA introgression inferred from multilocus genetic variation

Hybridization drives the evolutionary trajectory of many species or local populations, and assessing the geographic extent and genetic impact of interspecific gene flow may provide invaluable clues to understand population divergence or the adaptive relevance of admixture. In North America, hares (Lepus spp.) are key species for ecosystem dynamics and their evolutionary history may have been affected by hybridization. Here we reconstructed the speciation history of the three most widespread hares in North America – the snowshoe hare (Lepus americanus), the white‐tailed jackrabbit (L. townsendii) and the black‐tailed jackrabbit (L. californicus) – by analysing sequence variation at eight nuclear markers and one mitochondrial DNA (mtDNA) locus (6240 bp; 94 specimens). A multilocus–multispecies coalescent‐based phylogeny suggests that L. americanus diverged ~2.7 Ma and that L. californicus and L. townsendii split more recently (~1.2 Ma). Within L. americanus, a deep history of cryptic divergence (~2.0 Ma) was inferred, which coincides with major speciation events in other North American species. While the isolation‐with‐migration model suggested that nuclear gene flow was generally rare or absent among species or major genetic groups, coalescent simulations of mtDNA divergence revealed historical mtDNA introgression from L. californicus into the Pacific Northwest populations of L. americanus. This finding marks a history of past reticulation between these species, which may have affected other parts of the genome and influence the adaptive potential of hares during climate change.     

Molecular and morphological boundaries of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae)

Neoseiulus californicus (McGregor) is a natural enemy of pest mites used worldwide in many crops. Its correct identification is thus essential to ensure biological control success. The present study aimed to characterize molecular and morphological intraspecific variations for assisting in the diagnosis of the species and to build baseline information about expected variations within a commercially important phytoseiid species. Morphological and molecular [12S rRNA, cytochrome b mitochondrial (mt)DNA, and internal transcribed spacer] analyses were carried out on fourteen populations collected worldwide and on one mass‐reared strain. The genetic distances between the specimens of N. californicus and another related species were high and no overlap was observed, sustaining the reliability of such molecular methods for assisting a specific diagnosis. Furthermore, the genetic distances between populations of N. californicus were very low and overlap between intra‐ and interpopulations distances was observed, except for two populations collected in France (Marsillargues and Midi‐Pyrénées). The high mitochondrial differentiation between these two latter populations and the others questions their specific status: do they belong to the species N. californicus or to another cryptic species? However, using nuclear DNA marker analyses, no distinct differences were observed. Furthermore, even if significant morphological differences were observed between the populations, these differences were very small and the standard errors within each population were very low. We thus concluded that all the populations studied belong to the species N. californicus, despite unexpected high mitochondrial variations. The present study thus displays the importance of an integrative taxonomic approach for avoiding misidentifications. A discussion on morphological and mtDNA variations in relation to diagnostic reliability is developped. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 393–406.     

Development of polymorphic microsatellite markers for the sorghum plant bug, Stenotus rubrovittatus (Heteroptera: Miridae)

Nine polymorphic microsatellite DNA markers from the sorghum plant bug, Stenotus rubrovittatus, were isolated and characterized. These markers were used to analyse 22 individuals from a single field population. The number of alleles at these nine loci ranged from two to 28 (mean = 11.4) and heterozygosity ranged from 0.27 to 0.86 (mean = 0.58). Stenotus rubrovittatus has shown rapid population growth in the decades since the first report in the 1980s of serious damage to a rice crop. These microsatellite markers will be of value for studying both the population genetics and population dynamics of S. rubrovittatus.     

Mitochondrial DNA and RAPD polymorphisms in the haploid mite Brevipalpus phoenicis (Acari: Tenuipalpidae)

Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) is recognized as the vector of citrus leprosis virus that is a significant problem in several South American countries. Citrus leprosis has been reported from Florida in the past but no longer occurs on citrus in North America. The disease was recently reported in Central America, suggesting that B. phoenicis constitutes a potential threat to the citrus industries of North America and the Caribbean. Besides B. phoenicis, B. obovatus Donnadieu, and B. californicus (Banks) have been incriminated as vectors of citrus leprosis virus and each species has hundreds of host plants. In this study, Brevipalpus mite specimens were collected from different plants, especially citrus, in the States of Florida (USA) and São Paulo (Brazil), and reared on citrus fruit under standard laboratory conditions. Mites were taken from these colonies for DNA extraction and for morphological species identification. One hundred and two Random Amplified Polymorphic DNA (RAPD) markers were scored along with amplification and sequencing of a mitochondrial cytochrome oxidase subunit I gene fragment (374 bp). Variability among the colonies was detected with consistent congruence between both molecular data sets. The mites from the Florida and Brazilian colonies were morphologically identified as belonging to B. phoenicis, and comprise a monophyletic group. These colonies could be further diagnosed and subdivided geographically by mitochondrial DNA analysis.     

Integration of the predatory mite Neoseiulus californicus with petroleum spray oil treatments for control of Tetranychus marianae on eggplant

The red spider mite, Tetranychus marianae McGregor (Acari: Tetranychidae), a pest of several vegetable crops and perennials, is widespread in the Pacific Islands, like the Mariana Islands, where it was first reported. Recently, T. marianae has become a serious pest, particularly affecting eggplant. Efforts were therefore made to develop an integrated approach to control of T. marianae. First, indoor experiments on eggplant, Solanum melongena L. (Solanaceae), determined the optimal combination of petroleum spray oil (PSO; Volck® oil spray) with release of Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) for the management of T. marianae. Additionally, studies evaluated the viability of the predatory mites on Guam. The combination of N. californicus with PSOs produced significant control of T. marianae and did not affect the survival of N. californicus. Although dicofol and carbaryl reduced populations of T. marianae, they caused high mortality of N. californicus. Integrating PSO with N. californicus yielded better control of T. marianae than did N. californicus alone. The release of N. californicus at 200 individuals per plant reduced populations of T. marianae more than did other release rates. In addition, N. californicus was able to survive and become established after being released on Guam.     

Isolation and characterization of microsatellite loci in <Emphasis Type="Italic">Cryptocarya chinensis</Emphasis> in lower subtropical China

We report on the isolation and characterization of eight microsatellite markers from repetitive DNA enriched libraries for Cryptocarya chinensis from lower subtropical China. These are the first microsatellite loci reported for Cryptocarya species. The number of alleles ranged from three to nine. Observed and expected heterozygosities ranged from 0.0518 to 0.9910 and 0.5241 to 0.7935 for polymorphic loci, respectively. These markers will allow analyses of the baseline genetic variability and population structure of C. chinensis to enrich our scientific understanding of forest fragmentation on genetic health of this species and provide strategies for effective conservation and management in this area.     

Microsatellite DNA markers for the intertidal copepod Tigriopus californicus

Eleven microsatellite loci were isolated and characterized for the intertidal copepod Tigriopus californicus. Primers reliably amplify alleles in inbred lines derived from two divergent populations previously shown to have genetic distances of 18% for the mitochondrial cytochrome oxidase I gene. The 11 loci provided markers for eight of the species’ 12 chromosomes.     

Call Variation in the Bufo microscaphus Complex: Implications for Species Boundaries and the Evolution of Mate Recognition

Since the Modern Synthesis, many species concepts have considered characters mediating mate recognition essential to both the identification and definition of species. To explore divergence in mate recognition systems, calls were recorded and toads measured from all three species of the monophyletic Bufo microscaphus species complex including five populations of Bufo californicus, three populations of B. microscaphus, and one population of B. mexicanus. Call duration, dominant frequency, and pulse rate were significantly related to temperature, but not snout-vent length. When adjusted for temperature, calls of B. californicus had a significantly longer call duration, higher dominant frequency, and slower pulse rate than B. mexicanus and B. microscaphus, which did not differ from one another. However, the magnitude of variation among populations of B. californicus was similar to that between species. Discriminant analysis using call variables provided some separation of B. californicus and B. mexicanus from B. microscaphus, and cross-validation analysis correctly classified approximately 75% of B. californicus and B. mexicanus. Given only slight divergence in mate recognition systems between B. californicus and the other two taxa, the biological significance of this difference remains ambiguous. Comparisons among B. americanus group members indicate that hybridizing taxa may or may not exhibit divergence in advertisement calls. Maintenance of independently evolving lineages may be driven by other evolutionary mechanisms. These results support the notion that species recognition is best viewed as an effect of mate recognition.     

Characterization of new microsatellite loci from Vitis vinifera and their conservation in some Vitis species and hybrids

We present here characterization data for seven new microsatellite markers designed from new microsatellite loci isolated from a microsatellite‐enriched DNA library from Vitis vinifera. The observed heterozygosity varied from 0.73 up to 0.93 and the number of alleles per locus ranged from 12 to 26. This high polymorphism makes these new markers interesting for use in genotyping studies and completing the set of microsatellite markers already available for V. vinifera. Additionally these seven new markers appear to be conserved in four other Vitis species and 15 Vitis hybrids used as rootstocks for V. vinifera cultivation.     

Isolation and characterization of 22 polymorphic microsatellite DNA markers of Japanese sea bass (Laterolabrax japonicus)

Japanese sea bass (Laterolabrax japonicus) inhabits the coast of East Asia and is cage‐cultured currently in China as well. Twenty‐two microsatellite DNA markers were developed and used to type 35 individuals collected along the Chinese coast. The number of alleles per locus ranged from three to 23. The expected heterozygosity ranged from 0.111 to 0.938, while the observed heterozygosity ranged from 0.114 to 0.914. All 22 loci are neutral and independent of each other; two deviated significantly from Hardy–Weinberg equilibrium. These microsatellite DNA markers are moderately informative, which will certainly facilitate the management and exploitation of the genetic resource of L. japonicus.     

Biological control of twospotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis>, with predatory mite, <Emphasis Type="Italic">Neoseiulus californicus</Emphasis>, in strawberries

Greenhouse and field experiments were conducted from 2005 to 2007 to determine the effectiveness of different release times with the predatory mite, Neoseiulus californicus (McGregor), for control of the twospotted spider mite (TSSM), Tetranychus urticae Koch, in strawberries (Fragaria x ananassa Duchesne). The effect of N. californicus releases over time and on development of TSSM populations during a growing season were evaluated. Our hypothesis was that repeated applications of N. californicus, which is currently recommended by biological control companies, might be unnecessary to attain season-long control of TSSM. In greenhouse trials, three treatments consisting of releases of N. californicus at five-day intervals: day 0, day 5, and day 10, and an untreated control were evaluated. The treatment releases significantly reduced TSSM below the control within five days of each release. Neoseiulus californicus significantly reduced TSSM in treatments with high densities (leaflets with ≥ 40 TSSM) below that of treatments with lower densities (leaflets with ≤ 10 TSSM) demonstrating that if released at a predator: prey ratio of 1:10, timing of release does not alter the effectiveness of N. californicus in controlling TSSM. However, we found that if the ratio of predator: prey remains adequate, N. californicus is a more efficient predator at high TSSM densities. Field studies included three treatments consisting of releases of N. californicus at one-month intervals. All treatments significantly reduced TSSM compared with the control plots (no releases). Releases applied early in the season sustained TSSM significantly below those in the control plots for the whole season. Our results indicate that one release of N. californicus is able to sustained control of TSSM in strawberry throughout a growing season if released when TSSM populations are low early in the season in the southeastern United States.     

Development and characterization of 13 microsatellite markers for Cryptocarya concinna in lower subtropical China

We report on the development and characterization of 13 microsatellite markers from repetitive DNA enriched libraries for Cryptocarya concinna from lower subtropical China. The number of alleles ranged from 3 to 12. Observed and expected heterozygosities ranged from 0.4286 to 0.8571, and 0.4725 to 0.8820, respectively. These markers will allow analysis of the baseline genetic variability and population structure of C. concinna to enrich our scientific understanding of forest fragmentation on genetic health of this species and provide strategies for effective conservation and management in this area.     

Development of microsatellite markers for the drywood termite Incisitermes minor (Hagen)

Ten polymorphic microsatellite markers were developed in the drywood termite Incisitermes minor (Hagen) by using a genomic DNA extracted from the heads of workers. The microsatellite markers obtained in this study produced between two and seven alleles per locus. The observed and expected heterozygosities among the 10 markers ranged from 0.16 to 0.83 and from 0.43 to 0.84, respectively. These markers were shown to be good molecular tools for identification of the genetic structure and parentage assessment in I. minor.     

Polymorphic population genetic markers for the Australian wood cockroach Panesthia australis

Primers for five polymorphic nuclear DNA (nDNA) markers and one mitochondrial DNA (mtDNA) gene (COI) were developed for the Australian wood cockroach Panesthia australis using a range of techniques. Eight mitochondrial haplotypes and four to 36 alleles per nuclear locus were detected in 744 cockroaches. Observed heterozygosity ranged from 0.117 to 0.816 in a sample of 30 animals from one population. The markers will be useful for population biology and for the measurement of the effects of habitat fragmentation on this ecologically important forest‐dependent species.     

Characterization of Microsatellites in the IGF-2 and GH Genes of Asian Seabass (Lates calcarifer)

Four microsatellites were identified by screening the DNA sequences of Asian seabass (Lates calcarifer) deposited to GenBank. Two markers each are located in the growth hormone gene (GH) and in the insulin-like growth factor II gene (IGF-2), respectively. The markers were characterized by genotyping 34 Asian seabass individuals. All 4 microsatellites showed polymorphism: the number of alleles per locus ranged from 2 to 11 (average, 5.0), while the expected heterozygosity ranged from 0.51 to 0.85 (average, 0.63) at the 4 loci. Cross-priming with all 4 primer pairs was tested in species belonging to 5 different genera, but no bands were amplified. These microsatellites are the first genomic DNA markers characterized in L. calcarifer; thus they may be valuable for research and aquaculture production of this species. Received April 10, 2000; accepted July 13, 2000.