Isolation and characterization of microsatellite loci for the Asian longhorned beetle, Anoplophora glabripennis.

We describe the first isolation of SSR (simple sequence repeat) markers for Anoplophora glabripennis, a pest of forests in China and urban trees in the USA. Ten markers were developed using FIASCO (fast isolation by AFLP of sequences containing repeats) with an (AC)(17) probe, and five markers were identified in expressed sequence tags found on the NCBI database. Twenty-seven beetles from a population in Gansu Province, China, were assayed. The number of alleles ranged from 3 to 9; observed and expected heterozygosities ranged from 0.07 to 0.79 and from 0.30 to 0.78, respectively.

Challenges of detecting directional selection after a bottleneck: lessons from Sorghum bicolor.

Multilocus surveys of sequence variation can be used to identify targets of directional selection, which are expected to have reduced levels of variation. Following a population bottleneck, the signal of directional selection may be hard to detect because many loci may have low variation by chance and the frequency spectrum of variation may be perturbed in ways that resemble the effects of selection. Cultivated Sorghum bicolor contains a subset of the genetic diversity found in its wild ancestor(s) due to the combined effects of a domestication bottleneck and human selection on traits associated with agriculture. As a framework for distinguishing between the effects of demography and selection, we sequenced 204 loci in a diverse panel of 17 cultivated S. bicolor accessions. Genomewide patterns of diversity depart strongly from equilibrium expectations with regard to the variance of the number of segregating sites, the site frequency spectrum, and haplotype configuration. Furthermore, gene genealogies of most loci with an excess of low frequency variants and/or an excess of segregating sites do not show the characteristic signatures of directional and diversifying selection, respectively. A simple bottleneck model provides an improved but inadequate fit to the data, suggesting the action of other population-level factors, such as population structure and migration. Despite a known history of recent selection, we find little evidence for directional selection, likely due to low statistical power and lack of an appropriate null model.

Equilibrium processes cannot explain high levels of short- and medium-range linkage disequilibrium in the domesticated grass Sorghum bicolor.

Patterns of linkage disequilibrium (LD) are of interest because they provide evidence of both equilibrium (e.g., mating system or long-term population structure) and nonequilibrium (e.g., demographic or selective) processes, as well as because of their importance in strategies for identifying the genetic basis of complex phenotypes. We report patterns of short and medium range (up to 100 kb) LD in six unlinked genomic regions in the partially selfing domesticated grass, Sorghum bicolor. The extent of allelic associations in S. bicolor, as assessed by pairwise measures of LD, is higher than in maize but lower than in Arabidopsis, in qualitative agreement with expectations based on mating system. Quantitative analyses of the population recombination parameter, rho, however, based on empirical estimates of rates of recombination, mutation, and self-pollination, show that LD is more extensive than expected under a neutral equilibrium model. The disparity between rho and the population mutation parameter, , is similar to that observed in other species whose population history appears to be complex. From a practical standpoint, these results suggest that S. bicolor is well suited for association studies using reasonable numbers of markers, since LD typically extends at least several kilobases but has largely decayed by 15 kb.

MITE display.

Genome size differences among crop plants are largely due to unequal accumulation of repetitive DNA sequences, mainly transposable elements (TEs). Over the past decade, many families of miniature inverted-repeat transposable elements (MITEs) have been identified and characterized in a variety of organisms including animals and plants. MITEs are characterized by short terminal inverted repeats (TIRs) (10-15 bp), small size (approx 100 to 500 bp), high-copy-number (approx 1000 to 15,000 per haploid genome), and a preference for insertion into 2-bp to 3-bp targets that are rich in A and T residues. In this chapter, we present a modified transposon display procedure based on the maize MITE family Heartbreaker (Hbr). This technique is similar to AFLP in which AFLP adaptors are ligated to compatible ends of digested genomic DNA. Subsets of Hbr-containing fragments are then amplified using one AFLP primer and another primer complementary to an internal sequence of the Hbr element. Like AFLP, the Hbr display method permits the simultaneous analysis of numerous DNA fragments. Given the plethora of available marker systems, the major advantage of Hbr markers, and perhaps most MITE-based markers, is a preference for insertion in or near transcriptionally active genomic regions. This feature may be especially valuable in the large genomes of agriculturally important plants like maize, wheat, and barley where gene-rich islands are thought to exist in a sea of retrotransposons. Having a class of markers that are enriched in genic regions, coupled with the ease of isolating MITE markers, could expedite chromosome walks and map-based cloning protocols in these organisms.