Allison C Crawford

Genetic structuring in the spotted gum complex (genus Corymbia, section Politaria)

Spotted gums (genus Corymbia, section Politaria) occur as a species replacement series along the eastern seaboard of Australia, their distributions marked by regions of disjunction and sympatry. Their taxonomy remains controversial, with species assignment often challenging and reliant on knowledge of geographic origin as well as subtle morphological or leaf-oil variation. In the present paper, we explore a classification for spotted gums, without assuming predefined geographic or taxonomic groups but instead using genetic structure at microsatellite marker loci (n = 9) and a Bayesian model-based clustering approach implemented in Structure software. The C. torelliana outgroup (n = 21; section Cadagaria) formed a well resolved cluster (minimum pairwise Fst = 0.19). Four populations were evident within the spotted gums (n = 93) but structure was weak (pairwise Fst range 0.13–0.05). Geographic distance, topography and distribution disjunction were major determinants of structure, with migration among populations approximating a linear stepping-stone model. Corymbia maculata was resolved as a taxon and had the greatest genetic distance from any other population (minimum pairwise Fst 0.08). Three clusters were evident within the northern taxa but alignment with taxonomic groupings was poor. C. citriodora material from north of a major disjunction in central Queensland formed a Northern population. C. citriodora, C. variegata and C. henryi material south of this disjunction but north of the Border Range, formed a Central population, whereas a Southern population comprised C. variegata and C. henryi from predominately south of the Border Range.

Consistent production of cost-effective LongSAGE libraries

Serial analysis of gene expression (SAGE) and related techniques are gaining popularity as tools for exploring expression of plant genes but remain suboptimal because of smaller-than-expected average concatemer sizes. The presence of low-molecular-weight contaminants in high-molecular-weight concatemer fractions reduces the average size of cloned fragments, thereby limiting the viability of high-throughput sequencing methods. Implementation of an additional digestion step to promote formation of linear concatemer fragments appears to reduce the proportion of contaminants indirectly, but with variable results. We explored the effect of initial ditag polymerase chain reaction (PCR) quantity on the average size of cloned concatemers from the greater than 1000-bp fraction. The quantity of PCR material used was found to have a strong influence on the frequency of low-molecular-weight contaminants within this fraction, which has important implications for reducing costs associated with high-throughput sequencing of concatemer clones.