Xiao-Yue Hong

Invasion genetics of the Western flower thrips in China: evidence for genetic bottleneck, hybridization and bridgehead effect.

The western flower thrips, Frankliniella occidentalis (Pergande), is an invasive species and the most economically important pest within the insect order Thysanoptera. F. occidentalis, which is endemic to North America, was initially detected in Kunming in southwestern China in 2000 and since then it has rapidly invaded several other localities in China where it has greatly damaged greenhouse vegetables and ornamental crops. Controlling this invasive pest in China requires an understanding of its genetic makeup and migration patterns. Using the mitochondrial COI gene and 10 microsatellites, eight of which were newly isolated and are highly polymorphic, we investigated the genetic structure and the routes of range expansion of 14 F. occidentalis populations in China. Both the mitochondrial and microsatellite data revealed that the genetic diversity of F. occidentalis of the Chinese populations is lower than that in its native range. Two previously reported cryptic species (or ecotypes) were found in the study. The divergence in the mitochondrial COI of two Chinese cryptic species (or ecotypes) was about 3.3% but they cannot be distinguished by nuclear markers. Hybridization might produce such substantial mitochondrial-nuclear discordance. Furthermore, we found low genetic differentiation (global F STu200a=u200a0.043, P<0.001) among all the populations and strong evidence for gene flow, especially from the three southwestern populations (Baoshan, Dali and Kunming) to the other Chinese populations. The directional gene flow was further supported by the higher genetic diversity of these three southwestern populations. Thus, quarantine and management of F. occidentalis should focus on preventing it from spreading from the putative source populations to other parts of China.

Effects of Wolbachia on mtDNA variation and evolution in natural populations of Tetranychus urticae Koch.

We investigated the effects of Wolbachia infection on mtDNA variation in spider mites by sequencing a portion of the mitochondrial cytochrome oxidase I (COI) gene from 198 individuals of known infection status. Four Wolbachia strains were described in the current study, namely wUrtOri1, wUrtOri2, wUrtOri3 and wUrtCon1. As predicted, the haplotype and nucleotide diversity were lower in infected individuals than in uninfected individuals. However, these mtDNA haplotype data are not entirely concordant with the surface protein of wolbachia (wsp) sequence data and both infected and uninfected individuals were found of the same haplotype. Although values of Tajimas D and Fu & Lis F were consistently less than zero for most infected groups, McDonald–Kreitman tests suggested that the patterns of variation were different from those expected under neutrality in only the uninfected group. Thus, the neutrality tests do not show a clear effect of Wolbachia infection on patterns of mtDNA variation and substitution in spider mites.

Homoplastic evolution and host association of Eriophyoidea (Acari, Prostigmata) conflict with the morphological-based taxonomic system.

The superfamily Eriophyoidea is exceptionally diverse and its members are highly host-specific. Currently, the taxonomy of this group is based on morphology only. However, phylogenetic relationships in this group could be incorrect if the diagnostic morphological characters are homoplastic. Therefore, the phylogeny of 112 representative taxa of Eriophyoidea from China was determined using 18S, 28S D2-5 and D9-10 rRNA. Phylogenetic relationships were inferred through Bayesian, maximum likelihood and maximum parsimony methods, and then a number of clades or major clades were defined according to robust phylogenetic topologies combined with morphological comparison. Tests of monophyly showed that two of three families of Eriophyoidea as well as one subfamily and four tribes were not monophyletic. Ancestral character state reconstruction (ACSR) showed that five diagnostic morphological characters evolved several times, confounding the current taxonomy. Additionally, reconstruction of the history of host plant colonization suggested host switching occurred in a limited range of host plants. The host association data made it possible to determine taxonomic relationships more accurately. These results show that by integrating morphological and molecular information and host plant choice, it is possible to obtain a more accurate taxonomy and a deeper phylogenetic understanding of Eriophyoidea.

Mining and characterization of sequence tagged microsatellites from the brown planthopper Nilaparvata lugens.

Abstract n The brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), is an important pest of rice. To better understand the migration pattern and population structure of the Chinese populations of N. lugens, we developed and characterized 12 polymorphic microsatellites from the expressed sequence tags database of N. lugens. The occurrence of these simple sequence repeats was assessed in three populations collected from three provinces of China. The number of alleles per locus ranged from 3 to 13 with an average of 6.5 alleles per locus. The mean observed heterozygosity of the three populations ranged from 0.051 to 0.772 and the expected heterozygosity ranged from 0.074 to 0.766. The sequences of the 12 markers were highly variable. The polymorphism information content of the 12 markers was high and ranged from 0.074 to 0.807 (mean = 0.503). Sequencing of microsatellite alleles revealed that the fragment length differences were mainly due to the variation of the repeat motif. Significant genetic differentiation was detected among the three N. lugens populations as the Fst ranged from 0.034 to 0.273. Principle coordinates analysis also revealed significant genetic differentiation between populations of different years. We conclude that these microsatellite markers will be a powerful tools to study the migration routine of the N. lugens.

How do hosts react to endosymbionts? A new insight into the molecular mechanisms underlying the Wolbachia–host association

Wolbachia is an intracellular bacterium that has aroused intense interest because of its ability to alter the biology of its host in diverse ways. In the two‐spotted spider mite, Tetranychus urticae, Wolbachia can induce complex cytoplasmic incompatibility (CI) phenotypes and fitness changes, although little is known about the mechanisms. In the present study, we selected a strain of T.u2009urticae, in which Wolbachia infection was associated with strong CI and enhanced female fecundity, to investigate changes in the transcriptome of T.u2009urticae in Wolbachia‐infected vs. uninfected lines. The responses were found to be sex‐specific, with the transcription of 251 genes being affected in females and 171 genes being affected in males. Some of the more profoundly affected genes in both sexes were lipocalin genes and genes involved in oxidation reduction, digestion and detoxification. Several of the differentially expressed genes have potential roles in reproduction. Interestingly, unlike certain Wolbachia transinfections in novel hosts, the Wolbachia–host association in the present study showed no clear evidence of host immune priming by Wolbachia, although a few potential immune genes were affected.

The phylogenetic position of eriophyoid mites (superfamily Eriophyoidea) in Acariformes inferred from the sequences of mitochondrial genomes and nuclear small subunit (18S) rRNA gene.

Eriophyoid mites (superfamily Eriophyoidea) comprise >4400 species worldwide. Despite over a century of study, the phylogenetic position of these mites within Acariformes is still poorly resolved. Currently, Eriophyoidea is placed in the order Trombidiformes. We inferred the high-level phylogeny of Acari with the mitochondrial (mt) genome sequences of 110 species including four eriophyoid species, and the nuclear small subunit (18S) rRNA gene sequences of 226 species including 25 eriophyoid species. Maximum likelihood (ML), Bayesian inference (BI) and Maximum parsimony (MP) methods were used to analyze the sequence data. Divergence times were estimated for major lineages of Acari using Bayesian approaches. Our analyses consistently recovered the monophyly of Eriophyoidea but rejected the monophyly of Trombidiformes. The eriophyoid mites were grouped with the sarcoptiform mites, or were the sister group of sarcoptiform mites+non-eriophyoid trombidiform mites, depending on data partition strategies. Eriophyoid mites diverged from other mites in the Devonian (384Mya, 95% HPD, 352-410Mya). The origin of eriophyoid mites was dated to the Permian (262Mya, 95% HPD 230-307Mya), mostly prior to the radiation of gymnosperms (Triassic-Jurassic) and angiosperms (early Cretaceous). We propose that the placement of Eriophyoidea in the order Trombidiformes under the current classification system should be reviewed.

Cryptic diversity in host-associated populations of Tetra pinnatifidae (Acari: Eriophyoidea): what do morphometric, mitochondrial and nuclear data reveal and conceal?

Traditional morphology-based taxonomy of eriophyoid mites (Acari: Eriophyoidea) has been challenged by molecular-based technologies in the detection of cryptic species. However, the implications of such cryptic diversity appear to differ when methods based on different types of data are used. Here, samples of a host-associated eriophyoid mite species, Tetra pinnatifidae, collected from different host plants and localities are evaluated. The congruence of results based on morphometric (32 characters), mitochondrial (16S), and nuclear (28S) data were evaluated and showed a host-associated cryptic diversity dividing this morphospecies into several groups/clades that were morphometrically indistinguishable. In comparison, the 16S data confirmed cryptic speciation and intra-clade host-associated diversity, while 28S did not. In contrast, 28S data revealed potential gene flow between host-associated populations. High mitochondrial divergence, as well as low nuclear and morphological divergence indicated very recent stage of cryptic diversity of this eriophyoid mite.

Wolbachia-induced loss of male fertility is likely related to branch chain amino acid biosynthesis and iLvE in Laodelphax striatellus

Wolbachia are endosymbionts that infect many species of arthropods and nematodes. Wolbachia-induced cytoplasmic incompatibility (CI) is the most common phenotype in affected hosts, involving embryonic lethality in crosses between Wolbachia-infected males and uninfected females. The molecular mechanisms underlying this phenomenon are currently unclear. Here we examine the molecular correlates of the Wolbachia infection in Laodelphax striatellus (Fallén), an important rice pest, where embryonic lethality is strong and almost complete. We compared the gene expression of 4-day-old Wolbachia-infected and uninfected L.xa0striatellus testes to identify candidate genes for paternal-effect embryonic lethality induction. Based on microarray analysis, iLvE was the most down-regulated gene; this gene mediates branched-chain amino acid (BCAA) biosynthesis and participates in many processes related to reproductive performance. After knocking down iLvE by RNAi in uninfected male L.xa0striatellus, male fertility was reduced, leading to a decrease in embryo hatching rates, but fertility was rescued in crosses between these males and Wolbachia-infected females. Removal of BCAA in chemically-defined diets of uninfected males also led to a loss of male fertility. Low amino acid nutrition may enhance exposure time of sperm to Wolbachia in the testes to affect adult reproduction in L.xa0striatellus by reducing the number of sperm transferred per mating by males. These results indicate that Wolbachia may decrease male fertility in L.xa0striatellus by acting on iLvE, a key factor of BCAA biosynthesis, and delaying sperm maturation.

Sex-dependent activity of de novo methyltransferase 3 (Tudnmt3) in the two-spotted mite, Tetranychus urticae Koch

DNA methylation is an epigenetic mechanism for regulating developmental and other important processes in eukaryotes. Several components of the DNA methylation machinery have been identified, such as DNA methyltransferases. However, little is known about DNA methyltransferases in chelicerates, which is the second largest arthropod group. Epigenetics are expected to have a crucial role in the metabolism and development of this group. Here, we investigated the role of DNA methyltransferase 3 in the development of Tetranychus urticaeu2005Koch. In silico analyses clearly showed that this enzyme possesses the necessary conserved motifs for the catalytic activity of de novo methylation of DNA. Real‐time PCR revealed that T.u2009urticae de novo methyltransferase 3 (Tudnmt3) is expressed ubiquitously and throughout the life cycle of the two‐spotted spider mite. However, the pattern of Tudnmt3 expression was sex‐dependent during the adult stage. Whole in situ hybridization provided supportive evidence that Tudnmt3 is linked to the differentiation of the gonads in adult females and males. Methylation‐sensitive amplification polymorphism analyses of 119 loci showed that the status of DNA methylation is partially different between adult females and males, raising the possibility that this sex‐dependent DNA methylation pattern is mediated by different methylation activity of Tudnmt3.

Development of microsatellite markers for, and a preliminary population genetic analysis of, the white-backed planthopper

For a better understanding of the population structure and dispersal rates of Sogatella furcifera, we developed 21 novel polymorphic expressed sequence tags (EST) derived microsatellites, which were successfully amplified in four multiplex polymerase chain reaction sets. These new microsatellites were firstly assessed in 20 individuals sampled from Wenshan in China. The results showed that all 21 loci were highly polymorphic; the number of alleles ranged from 3 to 9, with an average of 4.8 alleles per locus. The observed and expected heterozygosity ranged from 0.200 to 0.900 and from 0.184 to 0.799, respectively. Nineteen of the 21 microsatellites without null allele, were subsequently used for population genetic structure analyses of five S. furcifera populations sampled in south region of China (sites up to 1314 kilometers apart). The observed and expected heterozygosity for each population ranged from 0.436 to 0.494 and from 0.454 to 0.482, respectively. The level of population differentiation was very low, with an average pairwise F ST of 0.002. Bayesian cluster analysis result suggested that the five S. furcifera populations formed one genetic cluster. Discriminant analysis of principle components detected three genetic clusters. The spread of the three clusters across the five populations explained the lack of population differentiation and the Bayesian cluster result. All the results indicated that long-distance migration of this pest allowed genetic mixing between populations from remote geographical origins. These new microsatellites will be powerful tools for population genetics studies of S. furcifera.