Agrobacterium-mediated transformation of Nicotiana attenuata, a model ecological expression system

Summary. Research into the genetic basis of the ecological sophistication of plants is hampered by the availability of transformable systems with a wealth of well-described ecological interactions. We present an Agrobacterium-mediated transformation system for the model ecological expression system, Nicotiana attenuata, a native tobacco that occupies the post-fire niche in the Great Basin Desert of North America. We describe a transformation vector and a transformation procedure that differs from the standard cultivated tobacco transformation protocols in its use of selectable markers, explants, media and cultivation conditions. We illustrate its utility in the transformations with genes coding for key enzymes in the oxylipin cascade (lipoxygenase, allene oxide synthase, hydroperoxide lyase) in antisense orientations and present high-throughput screens useful for the detection of altered phenotypes for the oxylipin cascade (green leaf volatiles and jasmonic acid after wounding). Received 14 March 2002; accepted 10 May 2002     

Plant-trait-based modeling assessment of ecosystem-service sensitivity to land-use change

Evidence is accumulating that the continued provision of essential ecosystem services is vulnerable to land-use change. Yet, we lack a strong scientific basis for this vulnerability as the processes that drive ecosystem-service delivery often remain unclear. In this paper, we use plant traits to assess ecosystem-service sensitivity to land-use change in subalpine grasslands. We use a trait-based plant classification (plant functional types, PFTs) in a landscape modeling platform to model community dynamics under contrasting but internally consistent land-use change scenarios. We then use predictive models of relevant ecosystem attributes, based on quantitative plant traits, to make projections of ecosystem-service delivery. We show that plant traits and PFTs are effective predictors of relevant ecosystem attributes for a range of ecosystem services including provisioning (fodder), cultural (land stewardship), regulating (landslide and avalanche risk), and supporting services (plant diversity). By analyzing the relative effects of the physical environment and land use on relevant ecosystem attributes, we also show that these ecosystem services are most sensitive to changes in grassland management, supporting current agri-environmental policies aimed at maintaining mowing of subalpine grasslands in Europe.     

Resource availability and the abundance of an N-based defense in Australian tropical rain forests

Plant defense theories predict that relatively resource-rich environments (those with more fertile soil) will support a greater abundance of plants with nitrogen-based chemical defense, but this has yet to be adequately tested. We tested this prediction by measuring the diversity and contribution to total biomass of cyanogenic plants (those that release hydrogen cyanide from endogenous cyanide-containing compounds) in the Australian tropical rain forest. We examined 401 species in thirty 200-m2 plots, six at each of five sites, for cyanogenesis. In upland/highland rain forest, two pairs of sites similar in rainfall and altitude, but differing in soil nutrients, were selected, as well as one site in lowland rain forest. Sites differed markedly in species composition and foliar N was positively related to soil fertility. Holding altitude constant, we did not detect significant differences in the proportion of cyanogenic species with soil fertility, nor did we consistently detect significant increases in the contribution of cyanogenic species to total biomass on higher nutrient sites. Thus we found no clear evidence that soil fertility affects the distribution and prevalence of species investing in a constitutive N-based defense at the community level.     

Habitat-mediated variation in predation risk by the American marten

The probability of prey encounter, attack, capture, and kill are often hypothesized to depend on habitat structure, but field evidence in terrestrial systems is rare. We tested whether predation efficiency by the American marten (Martes americana) and fear of predation by their primary prey, the red-backed vole (Clethrionomys gapperi), differed between 20- to 50-year-old regenerating forest stands and older uncut stands. Our results showed that the frequency of prey encounter, prey attack, and prey kill were higher in old uncut forests, despite the fact that small-mammal density was similar to that in younger logged forests. These differences in predation efficiency were linked to higher abundance of coarse woody debris, which seems to offer sensory cues to martens, thereby increasing the odds of hunting success. Red-backed voles in regenerating forest stands exhibited increased wariness compared to voles living in old uncut forest, suggestive of a behavioral response to habitat-mediated variation in predation risk.     

Plant growth rates and seed size: a re-evaluation

Small-seeded plant species are often reported to have high relative growth rate or RGR. However, because RGR declines as plants grow larger, small-seeded species could achieve higher RGR simply by virtue of their small size. In contrast, size-standardized growth rate or SGR factors out these size effects. Differences in SGR can thus only be due to differences in morphology, allocation, or physiology. We used nonlinear regression to calculate SGR for comparison with RGR for 10 groups of species spanning a wide range of life forms. We found that RGR was negatively correlated with seed mass in nearly all groups, but the relationship between SGR and seed mass was highly variable. We conclude that small-seeded species only sometimes possess additional adaptations for rapid growth over and above their general size advantage.     

Spatial distribution of defense chemicals and markers and the maintenance of chemical variation

Exploring the spatial distribution of variation in plant secondary metabolites is critical for understanding the evolutionary ecology of biochemical diversity in wild organisms. In the present study, concentrations of foliar sideroxylonal, an important and highly heritable defense chemical of Eucalyptus melliodora, displayed strong, fine-scale spatial autocorrelation. The spatial patterns observed could promote associational effects on herbivore foraging decisions, which may influence the selection pressures exerted on sideroxylonal content. Multiple chemical traits have roles in certain eucalypt-herbivore interactions, and the spatial characteristics of the herbivore foraging environment are therefore determined by these different factors. We used a model of E. melliodora intake by common brushtail possums (Trichosurus vulpecula), based on the combined effects of two chemical traits, to explore this idea and found that the spatial patterns were different to those of sideroxylonal alone. Spatial genetic autocorrelation, examined using microsatellites, was strong and occurred at a fine scale, implying that restricted gene flow might allow genetic patches to respond to selection relatively independently. Local two-dimensional genetic autocorrelation, explored using a new heuristic method, was highly congruent with the pattern of local phenotypic variation observed for sideroxylonal, suggesting that the genetic variance underlying the sideroxylonal variation is similarly structured. Our results suggest that the spatial distribution of genetic and phenotypic variation could influence both the selective pressure imposed by herbivores on eucalypt defenses and the potential of populations to respond to natural selection. Spatial context should be considered in future studies of plant-herbivore interactions.     

Using revealed preferences to estimate the Value of Travel Time to recreation sites

The opportunity Value of Travel Time (VTT) is one of the most important elements of the total cost of recreation day-trips and arguably the most difficult to estimate. Most studies build upon the theoretical framework proposed by Becker (1965) by using a combination of revealed and stated preference data to estimate a value of time which is uniform in all activities and under all circumstances. This restriction is relaxed by DeSerpa's (1971) model which allows the value of saving time to be activity-specific. We present the first analysis which uses actual driving choices between open access and toll roads to estimate a VTT specific for recreation trips, thereby providing a value which conforms to both Becker's and DeSerpa's theoretical models. Using these findings we conduct a Monte Carlo simulation to identify generalizable results for subsequent valuation studies. Our results indicate that 3/4 of the wage rate provides a reasonable approximation of the average VTT for recreation trips, while the commonly implemented assumption of 1/3 of the wage rate generates downward biased results.