Hannah McPherson and Maurizio Rossetto,
Key words: Swamp conservation, chloroplast DNA, genetic diversity, landscape connectivity
Introduction. Little is known about the historical or present-day connectivity of Temperate Highland Peat Swamps on Sandstone (THPSS) in the Sydney Basin (NSW). Recent technological advances have enabled exploration of genetic complexity at both species and community levels. By focusing on multiple plant species and populations, and investigating intraspecific gene-flow across multiple swamps, we can begin to make generalisations about how species and communities respond to change, thereby providing a solid scientific basis from which appropriate conservation and restoration strategies can be developed.
The study area comprised eight swamps distributed across four sites along an altitudinal gradient: Newnes (1200m); Leura (900m); Budderoo (600m); and Woronora (400m), see figure 1.
The aims were:
- To assess the relative genomic diversity among target species representing a range of life-history traits. This was achieved by sequencing chloroplast DNA and detecting variants in pooled samples from 25 species commonly occurring in swamps.
- To explore geographic patterns of diversity among swamps and across multiple species by designing targeted genomic markers and screening variants among populations within and between sites (for ten species occurring in up to 8 swamps).
- To develop a set of simple, effective and standardised tools for assessing diversity, connectivity and resilience of swamps to threats (from mining to climate change).
Our study comprises three main components:
1. Species-level assessment of genetic variation of swamp species
We have taken advantage of new available methods and technologies (McPherson et al. 2013 and The Organelle Assembler at http://pythonhosted.org/ORG.asm/) to sequence and assemble full chloroplast genomes of 20 plant species from swamps in the Sydney Basin and detect within and between-population variation. This enabled a rapid assessment of diversity among representatives of 12 families and a broad range of life-history traits – e.g. table 1. We are currently finalising our bioinformatic sampling of the data to ensure even coverage of chloroplast data across the species, however these preliminary data show that relative estimates are not a product of different amounts of chloroplast data retrieved (e.g. for the seven species with sequence length greater than 100,000 base pairs variation ranges from absent to high).
2. Swamp-level assessment of variation and connectivity using three target species – Baeckea linifolia (high diversity), Lepidosperma limicola (low diversity) and Boronia deanei subsp. deanei (restricted and threatened species).
From the initial species-level study we selected three very different species for detailed population-level studies. We designed markers to screen for variation within and among sites and explore landscape-level connectivity. We identified the Woronora Plateau as a possible refugium and we have uncovered interesting patterns of gene-flow on the Newnes Plateau. Two species, Lepidosperma limicola and Baeckea linifolia seem able to disperse over long distances while Boronia deanei subsp. deanei showed unexpected high levels of diversity despite very limited seed-mediated gene-flow between populations. Its current conservation status was supported by our findings. A unique pattern was found for each species, highlighting the need for a multispecies approach for understanding dynamics of this system in order to make informed decisions about, and plans for, conservation management.
3. Multi-species approach to assessing swamp community population dynamics
Since the population study approach proved successful we expanded our study to include population studies for a further ten species. This required development of new Next Generation Sequencing (NGS) approaches applicable to a wide range of study systems. This kind of approach will allow us to make informed generalisations about swamp communities for conservation management planning.
Table 1. Preliminary results showing relative chloroplast variation among 25 swamp species. Sequence length is in base pairs (bp) and relative level of variation was calculated as sequence length divided by number of variants to obtain an estimate of number of SNPs per base pair. Relative variation was then categorised as: High (one SNP every <1,000 bp); Moderate (one SNP every 1,000 – <5,000 bp); Low (one SNP every 5,000 – <10,000 bp); Very low (one SNP every >10,000 bp); or absent (no SNPs).
Results to date. We have assembled partial chloroplast genomes of 20 plant species from THPSS in the Sydney Basin and categorised relative measurements of diversity. Preliminary data from the three target species highlighted the need for multispecies studies and we are now finalizing our results from an expanded study (including 13 species) in order to better understand connectivity and resilience of THPSS and provide data critical for more informed conservation planning. We have produced unique, simple methods for assessing genetic diversity and understanding dynamics at both the species and site levels.
Lessons learned and future directions. We found that individual species have unique patterns of genetic variation that do not necessarily correspond with phylogeny or functional traits and thereby highlight the benefit of multispecies studies. We have developed a unique, simple method for screening for genetic variation across whole assemblages which can be applied to many study systems. Since our data capture and analysis methods are standardised it will be possible in the future to scale this work up to include more species and/or more geographic areas and analyse the datasets together to address increasingly complex research questions about the resilience of swamps in a changing landscape.
Stakeholders and Funding bodies. The following people have contributed to many aspects of this research, including design, fieldwork and data generation and analysis: Doug Benson and Joel Cohen (Royal Botanic Gardens and Domain Trust), Anthea Brescianini and Glenda Wardle (University of Sydney), David Keith (Office of Environment and Heritage).
This research was funded through the Temperate Highland Peat Swamps on Sandstone Research Program (THPSS Research Program). This Program was funded through an enforceable undertaking as per section 486A of the Environment Protection and Biodiversity Conservation Act 1999 between the Minister for the Environment, Springvale Coal Pty Ltd and Centennial Angus Place Pty Ltd. Further information on the enforceable undertaking and the terms of the THPSS Research Program can be found at www.environment.gov.au/news/2011/10/21/centennial-coal-fund-145-million-research-program.
Contact. Hannah McPherson, Biodiversity Research Officer, Royal Botanic Gardens and Domain Trust, Mrs Macquaries Road, Sydney 2000; Tel: +61292318181 Email: email@example.com