Sampling tissue for DNA analysis of trees: Trunk cambium as an alternative to canopy leaves

The number of studies of tropical tree species that use molecular tools is increasing, most of which collect leaf tissue for genomic DNA extraction. In tropical trees the canopy is not only frequently inaccessible, but also, once reached, the leaf tissue is often heavily defended against herbivory by high concentrations of anti-predation compounds, which may inhibit downstream applications, particularly PCR. Cambium tissue, accessed directly from the tree trunk at ground level, offers a readily accessible resource that is less hampered by the presence of defensive chemicals than leaf tissue. Here we describe a simple method for obtaining tissue from the cambial zone for DNA extraction and test the applicability of the method in a range of tropical tree species. The method was used successfully to extract DNA from 11 species in nine families. A subset of the DNA extracts was tested in more detail and proved to be highly suitable for AFLP analysis.

Chloroplast and total genomic diversity in the endemic Costa Rican tree Lonchocarpus costaricensis (J.D. Smith) Pittier (Papilionaceae).

In Mesoamerica, tropical dry forest is a highly threatened habitat, and species endemic to this environment are under extreme pressure. The tree species, Lonchocarpus costaricensis is endemic to the dry northwest of Costa Rica and southwest Nicaragua. It is a locally important species but, as land has been cleared for agriculture, populations have experienced considerable reduction and fragmentation. To assess current levels and distribution of genetic diversity in the species, a combination of chloroplast-specific (cpDNA) and whole genome DNA markers (amplified fragment length polymorphism, AFLP) were used to fingerprint 121 individual trees in 6 populations. Two cpDNA haplotypes were identified, distributed among populations such that populations at the extremes of the distribution showed lowest diversity. A large number (487) of AFLP markers were obtained and indicated that diversity levels were highest in the two coastal populations (Cobano, Matapalo, H = 0.23, 0.28 respectively). Population differentiation was low overall, F-ST = 0.12, although Matapalo was strongly differentiated from all other populations (F-ST = 0.16-0.22), apart from Cobano (F., = 0.11). Spatial genetic structure was present in both datasets at different scales: cpDNA was structured at a range-wide distribution scale, whilst AFLP data revealed genetic neighbourhoods on a population scale. In general, the habitat degradation of recent times appears not to have yet impacted diversity levels in mature populations. However, although no data on seed or saplings were collected, it seems likely that reproductive mechanisms in the species will have been affected by land clearance. It is recommended that efforts should be made to conserve the extant genetic resource base and further research undertaken to investigate diversity levels in the progeny generation.

Contrasting quantitative traits and neutral genetic markers for genetic resource assessment of Mesoamerican Cedrela odorata

We compared within-population variability and degree of population differentiation for neutral genetic markers (RAPDS) and eight quantitative traits in Central American populations of the endangered tree, Cedrela odorata. Whilst population genetic diversity for neutral markers (Shannon index) and quantitative traits (heritability, coefficient of additive genetic variation) were uncorrelated, both marker types revealed strong differentiation between populations from the Atlantic coast of Costa Rica and the rest of the species' distribution. The degree of interpopulation differentiation was higher for RAPD markers (F-ST 0.67 for the sampled Mesoamerican range) than for quantitative traits (Q(ST) = 0.30). Hence, the divergence in quantitative traits was lower than could have been achieved by genetic drift alone, suggesting that balancing selection for similar phenotypes in different populations of this species. Nevertheless, a comparison of pair-wise estimates of population differentiation in neutral genetic markers and quantitative traits revealed a strong positive correlation (r = 0.66) suggesting that, for C. odorata, neutral marker divergence could be used as a surrogate for adaptive gene divergence for conservation planning. The utility of this finding and suggested further work are discussed.

Regional and population-scale influences on genetic diversity partitioning within costa rican populations of the pioneer tree Vochysia ferruginea mart.

The neotropical pioneer species Vochysia ferruginea is locally important for timber and is being increasingly exploited. The sustainable utilisation of this species would benefit from an understanding of the level and partitioning of genetic diversity within remnant and secondary regrowth populations. We used data from total genome (amplified fragment length polymorphism, AFLP) and chloroplast genome markers to assay diversity levels within seven Costa Rican populations. Significant chloroplast differentiation between Atlantic and Pacific watersheds was observed, suggesting divergent historical origins for these populations. Contemporary gene flow, though extensive, is geographically constrained and a clear pattern of isolation by distance was detectable when an inter-population distance representing gene flow around the central Costa Rican mountain range was used. Overall population differentiation was low (F-ST = 0.15) and within-population diversity high, though variable (H-s=0.16-0.32), which fits with the overall pattern of population genetic structure expected for a widespread, outcrossed tropical tree. However genetic diversity was significantly lower and differentiation higher for recently colonised and disturbed populations compared to that at more established sites. Such a pattern seems indicative of a pioneer species undergoing repeated cycles of colonisation and succession.

Modeling the age of tropical moist forest fragments in heavily-cleared lowland landscapes of Colombia

Increasingly, large areas of native tropical forests are being transformed into a mosaic of human dominated land uses with scattered mature remnants and secondary forests. In general, at the end of the land clearing process, the landscape will have two forest components: a stable component of surviving mature forests, and a dynamic component of secondary forests of different ages. As the proportion of mature forests continues to decline, secondary forests play an increasing role in the conservation and restoration of biodiversity. This paper aims to predict and explain spatial and temporal patterns in the age of remnant mature and secondary forests in lowland Colombian landscapes. We analyse the age distributions of forest fragments, using detailed temporal land cover data derived from aerial photographs. Ordinal logistic regression analysis was applied to model the spatial dynamics of mature and secondary forest patches. In particular, the effect of soil fertility, accessibility and auto-correlated neighbourhood terms on forest age and time of isolation of remnant patches was assessed. In heavily transformed landscapes, forests account for approximately 8% of the total landscape area, of which three quarters are comprised of secondary forests. Secondary forest growth adjacent to mature forest patches increases mean patch size and core area, and therefore plays an important ecological role in maintaining landscape structure. The regression models show that forest age is positively associated with the amount of neighbouring forest, and negatively associated with the amount of neighbouring secondary vegetation, so the older the forest is the less secondary vegetation there is adjacent to it. Accessibility and soil fertility also have a negative but variable influence on the age of forest remnants. The probability of future clearing if current conditions hold is higher for regenerated than mature forests. The challenge of biodiversity conservation and restoration in dynamic and spatially heterogeneous landscape mosaics composed of mature and secondary forests is discussed. (c) 2004 Elsevier B.V. All rights reserved.

A new method for conservation planning for the persistence of multiple species

Although the aim of conservation planning is the persistence of biodiversity, current methods trade-off ecological realism at a species level in favour of including multiple species and landscape features. For conservation planning to be relevant, the impact of landscape configuration on population processes and the viability of species needs to be considered. We present a novel method for selecting reserve systems that maximize persistence across multiple species, subject to a conservation budget. We use a spatially explicit metapopulation model to estimate extinction risk, a function of the ecology of the species and the amount, quality and configuration of habitat. We compare our new method with more traditional, area-based reserve selection methods, using a ten-species case study, and find that the expected loss of species is reduced 20-fold. Unlike previous methods, we avoid designating arbitrary weightings between reserve size and configuration; rather, our method is based on population processes and is grounded in ecological theory.

Non-linear principal components analysis: an alternative method for finding patterns in environmental data

The main purpose of this article is to gain an insight into the relationships between variables describing the environmental conditions of the Far Northern section of the Great Barrier Reef, Australia, Several of the variables describing these conditions had different measurement levels and often they had non-linear relationships. Using non-linear principal component analysis, it was possible to acquire an insight into these relationships. Furthermore. three geographical areas with unique environmental characteristics could be identified. Copyright (c) 2005 John Wiley & Sons, Ltd.

Significant patterns of population genetic structure and limited gene flow in a threatened macropodid marsupial despite continuous habitat in southeast Queensland, Australia

Many endangered species worldwide are found in remnant populations, often within fragmented landscapes. However, when possible, an understanding of the natural extent of population structure and dispersal behaviour of threatened species would assist in their conservation and management. The brush-tailed rock-wallaby (Petrogale penicillata), a once abundant and widespread rock-wallaby species across southeastern Australia, has become nearly extinct across much of the southern part of its range. However, the northern part of the species' range still sustains many small colonies closely distributed across suitable habitat, providing a rare opportunity to investigate the natural population dynamics of a listed threatened species. We used 12 microsatellite markers to investigate genetic diversity, population structure and gene flow among brush-tailed rock-wallaby colonies within and among two valley regions with continuous habitat in southeast Queensland. We documented high and signifcant levels of population genetic structure between rock-wallaby colonies embedded in continuous escarpment habitat and forest. We found a strong and significant pattern of isolation-by-distance among colonies indicating restricted gene flow over a small geographic scale (< 10 km) and conclude that gene flow is more likely limited by intrinsic factors rather than environmental factors. In addition, we provide evidence that genetic diversity was significantly lower in colonies located in a more isolated valley region compared to colonies located in a valley region surrounded by continuous habitat. These findings shed light on the processes that have resulted in the endangered status of rock-wallaby species in Australia and they have strong implications for the conservation and management of both the remaining 'connected' brush-tailed rock-wallaby colonies in the northern parts of the species' range and the remnant endangered populations in the south.

Effects of fire on the structure and composition of open eucalypt forests

Fires are integral to the healthy functioning of most ecosystems and are often poorly understood in policy and management, however, the relationship between floristic composition and habitat structure is intrinsically linked, particularly after fire. The aim of this study was to test whether the variability of habitat structure or floristic composition and abundance in forests at a regional scale can be explained in terms of fire frequency using historical data and experimental prescribed burns. We tested this hypothesis in open eucalypt forests of Fraser Island off the east coast of Australia. Fraser Island dunes show progressive stages in plant succession as access to nutrients decreases across the Island. We found that fire frequency was not a good predictor of floristic composition or abundance across dune systems; rather, its affects were dune specific. In contrast, habitat structure was strongly influenced by fire frequency, independent of dune system. A dense understorey occurred in frequently burnt areas, whereas infrequently burnt areas had a more even distribution of plant heights. Plant communities returned to pre-burn levels of composition and abundances within 6 months of a fire and frequently burnt areas were dominated by early successional species of plant. These ecosystems were characterized by low diversity and frequently burnt areas on the east coast were dominated by Pteridium. Greater midstorey canopy cover in low frequency areas reduces light penetration and allows other species to compete more effectively with Pteridium. Our results strongly indicate that frequent fires on the Island have resulted in a decrease in relative diversity through dominance of several species. Prescribed fire represents a powerful management tool to shape habitat structure and complexity of Fraser Island forests.

Precipitation and Fire Effects on Flowering of a Rare Prairie Orchid

A small, isolated population of the threatened western prairie fringed orchid (Platanthera praeclara Sheviak & Bowles) occurs at Pipestone National Monument, Minnesota, in a mesic prairie that is periodically burned to control invasive cool-season grasses. During 1995-2004, monitoring counts of flowering orchids in the monument varied considerably for different years. Similar precipitation amounts in the spring and histories of burning suggest that fire and precipitation in the spring were not the causes of the variation. For the eight non-burn years in the monitoring record, we compared the number of flowering plants and the precipitation amounts during six growth stages of the orchid and found a 2-variab1e model (precipitation during senescence/bud development and precipitation in the dormant period) explained 77% of the annual variation in number of flowering plants. We also conducted a fire experiment in early May 2002, the typical prescribed burn period for the monument, and found that the frequency of flowering, vegetative, and absent plants observed in July did not differ between burned and protected locations of orchids. We used the model and forecasts of precipitation in the spring to develop provisional burn decision scenarios. We discussed management implications of the scenarios.

Reoperative parathyroid surgery: The importance of ectopic location and multigland disease

Background: Despite the availability of expert surgeons and preoperative imaging investigations, some patients require reoperation for persistent or recurrent hyperparathyroidisms. Method: Fifty consecutive patients were reviewed. Results: There were 28 persistent cases (24 primary, 4 secondary) and 22 recurrent cases (15 primary, 7 secondary) and 98% had successful surgical treatment. Multigland disease was present in 24 of 39 (62%) of primary cases, 11 of 24 persistent and 13 of 15 recurrent (P < 0.02). Four patients in the recurrent primary group had multiple endocrine neoplasia type 1, whereas the other 20 primary patients had sporadic multigland disease. Multigland disease was present in all secondary cases and was a very important factor in this entire series of patients (70%). Regrowth of a remnant of a gland biopsied or partially resected at an earlier operation was the cause of recurrence in 12 of 15 primary and 2 of 7 secondary cases (P < 0.05). The site of missed glands in persistent disease was ectopic in 60%. Ectopic glands were found in the following sites: intrathyroidal 10 (8 inferior and 2 superior), intrathymic 9, posterior mediastinum 4, base of skull 2, carotid sheath 1 and supernumerary 5. Investigations to locate missing glands were positive in 28 of 43 sestamibi scans (65%), 14 of 34 ultrasound scans (41%), 10 of 24 computed tomography scans (42%) and 11 of 13 selective venous sampling tests (85%). Conclusion: Some persistent cases are unavoidable because of ectopic locations and some recurrences are inevitable because of multigland disease.

Review of the ecology of Australian urban fauna: A focus on spatially explicit processes

Cities have a major impact on Australian landscapes, especially in coastal regions, to the detriment of native biodiversity. Areas suitable for urban development often coincide with those areas that support high levels of species diversity and endemism. However, there is a paucity of reliable information available to guide urban conservation planning and management, especially regarding the trade-off between investing in protecting and restoring habitat at the landscape level, and investing in programmes to maintain the condition of remnant vegetation at the local (site) level. We review the literature on Australian urban ecology, focusing on urban terrestrial and aquatic vertebrate and invertebrate fauna. We identify four main factors limiting our knowledge of urban fauna: (i) a lack of studies focusing at multiple ecological levels; (ii) a lack of multispecies studies; (iii) an almost total absence of long-term (temporal) studies; and (iv) a need for stronger integration of research outcomes into urban conservation planning and management. We present a set of key principles for the development of a spatially explicit, long-term approach to urban fauna research. This requires an understanding of the importance of local-level habitat quality and condition relative to the composition, configuration and connectivity of habitats within the larger urban landscape. These principles will ultimately strengthen urban fauna management and conservation planning by enabling us to prioritize and allocate limited financial resources to maximize the conservation return.

The importance of forest area and configuration relative to local habitat factors for conserving forest mammals: A case study of koalas in Queensland, Australia

The loss and fragmentation of forest habitats by human land use are recognised as important factors influencing the decline of forest-dependent fauna. Mammal species that are dependent upon forest habitats are particularly sensitive to habitat loss and fragmentation because they have highly specific habitat requirements, and in many cases have limited ability to move through and utilise the land use matrix. We addressed this problem using a case study of the koala (Phascolarctos cinereus) surveyed in a fragmented rural-urban landscape in southeast Queensland, Australia. We applied a logistic modelling and hierarchical partitioning analysis to determine the importance of forest area and its configuration relative to site (local) and patch-level habitat variables. After taking into account spatial auto-correlation and the year of survey, we found koala occurrence increased with the area of all forest habitats, habitat patch size and the proportion of primary Eucalyptus tree species; and decreased with mean nearest neighbour distance between forest patches, the density of forest patches, and the density of sealed roads. The difference between the effect of habitat area and configuration was not as strong as theory predicts, with the configuration of remnant forest becoming increasingly important as the area of forest habitat declines. We conclude that the area of forest, its configuration across the landscape, as well as the land use matrix, are important determinants of koala occurrence, and that habitat configuration should not be overlooked in the conservation of forest-dependent mammals, such as the koala. We highlight the implications of these findings for koala conservation. (c) 2006 Elsevier Ltd. All rights reserved.