Western Australian sandalwood, Santalum spicatum, is widespread in the semi-arid and arid regions of Western Australia, and there is some morphological variation suggestive of two ecotypes. The level and structuring of genetic diversity within the species was investigated using anonymous nuclear RFLP loci. Santalum spicatum showed moderate levels of genetic diversity compared to other Australian tree species. The northern populations in the arid region showed greater levels of diversity and less population differentiation than the southern populations in the semi-arid region due to differences in the distribution of rare alleles. Equilibrium between drift and gene flow in the northern populations indicated that they have been established for a long period of time with stable conditions conducive to gene flow. In contrast, the southern populations showed a relationship between drift and gene flow indicative of a pattern of fragmentation and isolation where drift has greater effect than gene flow. The different patterns of diversity suggest that the ecotypes in the two regions have been subject to differences in the relative influences of drift and gene flow during their evolutionary history.
The Acacia acuminata complex includes three taxa, A. acuminata ssp. acuminata, A. acuminata ssp. burkittii and A. oldfieldii, along with several informal variants of A. acuminata. It is widespread throughout southern Australia with the centre of diversity in south-west Western Australia. Phylogeographical patterns in the complex were investigated using a nested clade analysis of cpDNA RFLPs from 25 populations in Western Australia. Except for A. oldfieldii that was clearly identified as a distinct entity, haplotypes were not restricted to sub-specific taxa or variants within A. acuminata. There was significant association between phylogenetic position of many haplotypes and their geographical distribution. The fine-scale phylogeographical patterns were complex but at deeper levels in the phylogeny there was evidence of divergence between two lineages. The pattern of shared haplotypes between lineages suggests retention of ancestral polymorphism as a result of incomplete lineage sorting. The divergence of these lineages is consistent with fragmentation caused by climatic instability during the Pleistocene.
The influence of geographic range on species persistence has long been of interest and there is a need for a better understanding of the genetic consequences for species with restricted distributions, particularly with the increasing rate of global species extinctions. However, the genetic effects of restricted range are often confounded by the impacts of population distribution. We compared chloroplast and nuclear genetic diversity and differentiation in two acacias, the restricted, patchily distributed Acacia atkinsiana and the widespread, semi-continuously distributed A. ancistrocarpa. Lower intra-population diversity and higher differentiation between populations were seen in A. atkinsiana compared to its widespread congener, A. ancistrocarpa. There was little evidence of geographical influences on population genetic structure in A. ancistrocarpa whereas A. atkinsiana exhibited nuclear genetic structure with isolation by distance, differentiation of near-coastal populations from those in the ranges, and differentiation of peripheral populations from those in the centre of the distribution. These results are consistent with expectations of the effect of geographic range and population distribution on genetic diversity, but indicate that distribution of populations rather than geographic range has influenced the observed genetic structure. The contrasting patterns observed here demonstrate that conservation approaches for species management and ecological restoration need to consider the distribution of populations in geographically restricted species.
Polyethylenimines (PEIs) of a molecular weight between 25 and about 800 kDa have successfully been used for in vitro and in vivo gene delivery approaches. Recent publications indicated that PEI molecules of lower molecular weight and a small molecular weight range are also efficient transfection reagents with a much lower cytotoxicity compared to high molecular weight PEIs. Here, we describe the application of a molecular sieve chromatography to fractionate a commercially available 25-kDa PEI. We generated three pools of PEIs with molecular weight ranges of 70-360 (I), 10-70 (II), and 0.5-10 kDa (III), respectively. We show that, in comparison with the 25-kDa PEI, pool III increased the expression of luciferase up to 100-fold and the number of transfected cells 2-3-fold. In addition, the kinetics of reporter gene expression was also much faster in pool III, compared with the 25-kDa PEI or with pools I or II. Finally, pool III showed the lowest cytotoxicity in comparison with the other PEI preparations. Thus, we provide a one-step processing of a 25-kDa PEI, resulting in a more effective and also less cytotoxic transfection reagent.
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