What causes female bias in the secondary sex ratios of the dioecious woody shrub Salix sitchensis colonizing a primary successional landscape?

UNLABELLED: • PREMISE OF THE STUDY: Females often outnumber males in Salix populations, although the mechanisms behind female bias are not well understood and could be caused by both genetic and ecological factors. We investigated several ecological factors that could bias secondary sex ratios of Salix sitchensis colonizing Mount St. Helens after the 1980 eruption.• METHODS: We determined whether S. sitchensis secondary sex ratios varied across disturbance zones created by the eruption and across mesic and hydric habitats within each zone. For one population, we tracked adult mortality, whole-plant reproductive allocation, the number of stems, and plant size for 2 years. In a field experiment, we created artificial streams to test whether vegetative reproduction via stem fragments was sex-biased.• KEY RESULTS: We found a consistent 2:1 female bias in S. sitchensis secondary sex ratios across all disturbance zones and habitats. Despite female plants sometimes allocating more resources (in terms of carbon, nitrogen, and phosphorus) to reproduction than males, we found no evidence of sex-biased mortality. The establishment rate of S. sitchensis experimental stems did not differ between the sexes, indicating that vegetative reproduction was not distorting secondary sex ratios.• CONCLUSIONS: We hypothesize that S. sitchensis secondary sex ratios depend on either early-acting genetic factors affecting the seed sex ratio or sex-specific germination or survival rates before maturity, as opposed to factors associated with reproduction in adult plants.

Gopher mounds decrease nutrient cycling rates and increase adjacent vegetation in volcanic primary succession.

Fossorial mammals may affect nutrient dynamics and vegetation in recently initiated primary successional ecosystems differently than in more developed systems because of strong C and N limitation to primary productivity and microbial communities. We investigated northern pocket gopher (Thomomys talpoides) effects on soil nutrient dynamics, soil physical properties, and plant communities on surfaces created by Mount St. Helens' 1980 eruption. For comparison to later successional systems, we summarized published studies on gopher effects on soil C and N and plant communities. In 2010, 18 years after gopher colonization, we found that gophers were active in ~2.5% of the study area and formed ~328 mounds ha(-1). Mounds exhibited decreased species density compared to undisturbed areas, while plant abundance on mound margins increased 77%. Plant burial increased total soil carbon (TC) by 13% and nitrogen (TN) by 11%, compared to undisturbed soils. Mound crusts decreased water infiltration, likely explaining the lack of detectable increases in rates of NO3-N, NH4-N or PO4-P leaching out of the rooting zone or in CO2 flux rates. We concluded that plant burial and reduced infiltration on gopher mounds may accelerate soil carbon accumulation, facilitate vegetation development at mound edges through resource concentration and competitive release, and increase small-scale heterogeneity of soils and communities across substantial sections of the primary successional landscape. Our review indicated that increases in TC, TN and plant density at mound margins contrasted with later successional systems, likely due to differences in physical effects and microbial resources between primary successional and older systems.

Phosphorus-mobilization ecosystem engineering: the roles of cluster roots and carboxylate exudation in young P-limited ecosystems.

BACKGROUND: Carboxylate-releasing cluster roots of Proteaceae play a key role in acquiring phosphorus (P) from ancient nutrient-impoverished soils in Australia. However, cluster roots are also found in Proteaceae on young, P-rich soils in Chile where they allow P acquisition from soils that strongly sorb P. SCOPE: Unlike Proteaceae in Australia that tend to proficiently remobilize P from senescent leaves, Chilean Proteaceae produce leaf litter rich in P. Consequently, they may act as ecosystem engineers, providing P for plants without specialized roots to access sorbed P. We propose a similar ecosystem-engineering role for species that release large amounts of carboxylates in other relatively young, strongly P-sorbing substrates, e.g. young acidic volcanic deposits and calcareous dunes. Many of these species also fix atmospheric nitrogen and release nutrient-rich litter, but their role as ecosystem engineers is commonly ascribed only to their diazotrophic nature. CONCLUSIONS: We propose that the P-mobilizing capacity of Proteaceae on young soils, which contain an abundance of P, but where P is poorly available, in combination with inefficient nutrient remobilization from senescing leaves allows these species to function as ecosystem engineers. We suggest that diazotrophic species that colonize young soils with strong P-sorption potential should be considered for their positive effect on P availability, as well as their widely accepted role in nitrogen fixation. Their P-mobilizing activity possibly also enhances their nitrogen-fixing capacity. These diazotrophic species may therefore facilitate the establishment and growth of species with less-efficient P-uptake strategies on more-developed soils with low P availability through similar mechanisms. We argue that the significance of cluster roots and high carboxylate exudation in the development of young ecosystems is probably far more important than has been envisaged thus far.

A stoichiometric model of early plant primary succession.

The relative importance of plant facilitation and competition during primary succession depends on the development of ecosystem nutrient pools, yet the interaction of these processes remains poorly understood. To explore how these mechanisms interact to drive successional dynamics, we devised a stoichiometric ecosystem-level model that considers the role of nitrogen and phosphorus limitation in plant primary succession. We applied this model to the primary plant community on Mount St. Helens, Washington State, to check the validity of the proposed mechanisms. Our results show that the plant community is colimited by nitrogen and phosphorus, and they confirm previous suggestions that the presence of a nitrogen-fixing legume, Lupinus lepidus, can enhance community biomass. In addition, the observed nutrient supply rates may promote alternative successional trajectories that depend on the initial plant abundances, which may explain the observed heterogeneity in community development. The model further indicates the importance of mineralization rates and other ecosystem parameters to successional rates. We conclude that a model framework based on ecological stoichiometry allows integration of key biotic processes that interact nonlinearly with biogeochemical aspects of succession. Extension of this approach will improve the understanding of the process of primary succession and its application to ecosystem rehabilitation.

Structure of the glucanase inhibitor protein (GIP) family from phytophthora species suggests coevolution with plant endo-beta-1,3-glucanases.

During invasion of their plant hosts, species of the oomycete genus Phytophthora secrete glucanase inhibitor proteins (GIPs) into the plant apoplast, which bind and inhibit the activity of plant extracellular endo-beta-1,3-glucanases (EGases). GIPs show structural homology to the chymotrypsin class of serine proteases (SP) but lack proteolytic activity due to the absence of an intact catalytic triad and, thus, belong to a broader class of proteins called serine protease homologs (SPH). To study the evolutionary relationship between GIPs and functional SP, database searches were used to identify 48 GIP homologs in the P. sojae, P. ramorum, and P. infestans genomes, composing GIPs, SPH, and potentially functional SP. Analyses of P. infestans-inoculated tomato leaves showed that P. infestans GIPs and tomato EGases are present in the apoplast and form stable complexes in planta. Studies of the temporal expression of a four-membered GIP family from P. infestans (PiGIP1 to PiGIP4) further revealed that the genes show distinctly different patterns during an infection timecourse. Codon evolution analyses of GIP homologs identified several positively selected peptide sites and structural modeling revealed them to be in close proximity to rapidly evolving EGase residues, suggesting that the interaction between GIPs and EGases has the hallmarks of a coevolving molecular arms race.

When can herbivores slow or reverse the spread of an invading plant? A test case from Mount St. Helens.

Here we study the spatial dynamics of a coinvading consumer-resource pair. We present a theoretical treatment with extensive empirical data from a long-studied field system in which native herbivorous insects attack a population of lupine plants recolonizing a primary successional landscape created by the 1980 volcanic eruption of Mount St. Helens. Using detailed data on the life history and interaction strengths of the lupine and one of its herbivores, we develop a system of integrodifference equations to study plant-herbivore invasion dynamics. Our analyses yield several new insights into the spatial dynamics of coinvasions. In particular, we demonstrate that aspects of plant population growth and the intensity of herbivory under low-density conditions can determine whether the plant population spreads across a landscape or is prevented from doing so by the herbivore. In addition, we characterize the existence of threshold levels of spatial extent and/or temporal advantage for the plant that together define critical values of "invasion momentum," beyond which herbivores are unable to reverse a plant invasion. We conclude by discussing the implications of our findings for successional dynamics and the use of biological control agents to limit the spread of pest species.

Predation on the Invasive Copepod, Pseudodiaptomus forbesi, and Native Zooplankton in the Lower Columbia River: An Experimental Approach to Quantify Differences in Prey-Specific Feeding Rates.

Invasive planktonic crustaceans have become a prominent feature of aquatic communities worldwide, yet their effects on food webs are not well known. The Asian calanoid copepod, Pseudodiaptomus forbesi, introduced to the Columbia River Estuary approximately 15 years ago, now dominates the late-summer zooplankton community, but its use by native aquatic predators is unknown. We investigated whether three species of planktivorous fishes (chinook salmon, three-spined stickleback, and northern pikeminnow) and one species of mysid exhibited higher feeding rates on native copepods and cladocerans relative to P. forbesi by conducting `single-prey' feeding experiments and, additionally, examined selectivity for prey types with `two-prey' feeding experiments. In single-prey experiments individual predator species showed no difference in feeding rates on native cyclopoid copepods (Cyclopidae spp.) relative to invasive P. forbesi, though wild-collected predators exhibited higher feeding rates on cyclopoids when considered in aggregate. In two-prey experiments, chinook salmon and northern pikeminnow both strongly selected native cladocerans (Daphnia retrocurva) over P. forbesi, and moreover, northern pikeminnow selected native Cyclopidae spp. over P. forbesi. On the other hand, in two-prey experiments, chinook salmon, three-spined stickleback and mysids were non- selective with respect to feeding on native cyclopoid copepods versus P. forbesi. Our results indicate that all four native predators in the Columbia River Estuary can consume the invasive copepod, P. forbesi, but that some predators select for native zooplankton over P. forbesi, most likely due to one (or both) of two possible underlying casual mechanisms: 1) differential taxon-specific prey motility and escape responses (calanoids > cyclopoids > daphnids) or 2) the invasive status of the zooplankton prey resulting in naivety, and thus lower feeding rates, of native predators feeding on invasive prey.

Trophic Interactions during Primary Succession: Herbivores Slow a Plant Reinvasion at Mount St. Helens.

Lupines (Lupinus lepidus var. lobbii), the earliest plant colonists of primary successional habitats at Mount St. Helens, were expected to strongly affect successional trajectories through facilitative effects. However, their effects remain localized because initially high rates of reinvasive spread were short lived, despite widespread habitat availability. We experimentally tested whether insect herbivores, by reducing plant growth and fecundity at the edge of the expanding lupine population, could curtail the rate of reinvasion and whether those herbivores had comparable impacts in the older, more successionally advanced core region. We found that removing insect herbivores increased both the areal growth of individual lupine plants and the production of new plants in the edge region, thereby accelerating the lupine's intrinsic rate of increase at the front of the lupine reinvasion. We found no such impacts of herbivory in the core region, where low plant quality or a complex of recently arrived natural enemies may hold herbivores in check. In the context of invasion theory, herbivore-mediated decreases in lupine population growth rate in the edge region translate into decreased rates of lupine spread, which we quantify here using diffusion models. In the Mount St. Helens system, decreased rate of lupine reinvasion will result in reductions in rates of soil formation, nitrogen input, and entrapment of seeds and detritus that are likely to postpone or alter trajectories of primary succession. If the type of spatial subtleties in herbivore effects we found here are common, with herbivory focused on the edge of an expanding plant population and suppressed or ineffective in the larger, denser central region (where the plants might be more readily noticed and studied), then insect herbivores may have stronger impacts on the dynamics of primary succession and plant invasions than previously recognized.

The effect of consumers and mutualists of Vaccinium membranaceum at Mount St. Helens: dependence on successional context.

In contrast to secondary succession, studies of terrestrial primary succession largely ignore the role of biotic interactions, other than plant facilitation and competition, despite the expectation that simplified interaction webs and propagule-dependent demographics may amplify the effects of consumers and mutualists. We investigated whether successional context determined the impact of consumers and mutualists by quantifying their effects on reproduction by the shrub Vaccinium membranaceum in primary and secondary successional sites at Mount St. Helens (Washington, USA), and used simulations to explore the effects of these interactions on colonization. Species interactions differed substantially between sites, and the combined effect of consumers and mutualists was much more strongly negative for primary successional plants. Because greater local control of propagule pressure is expected to increase successional rates, we evaluated the role of dispersal in the context of these interactions. Our simulations showed that even a small local seed source greatly increases population growth rates, thereby balancing strong consumer pressure. The prevalence of strong negative interactions in the primary successional site is a reminder that successional communities will not exhibit the distribution of interaction strengths characteristic of stable communities, and suggests the potential utility of modeling succession as the consequence of interaction strengths.

Resource use efficiency and community effects of invasive Hypochaeris radicata (Asteraceae) during primary succession.

PREMISE OF THE STUDY: We sought to better understand the impacts and mechanisms underpinning a successful invasion of resource-poor sites by a nonnative plant on Mount St. Helens volcano (MSH). • METHODS: We investigated the short-term effects of the nonnative plant Hypochaeris radicata on growth of native species colonizing drought-prone primary successional surfaces under N-limited and N-augmented conditions. To understand the success of H. radicata, we compared its resource use efficiency to that of a closely related native colonist, Hieracium albiflorum, under the same conditions. • KEY RESULTS: Removing H. radicata did not affect growth of the most common colonists, but N addition demonstrated strong N limitation to growth in H. albiflorum, H. radicata, and Agrostis spp. Nonnative H. radicata exhibited lower water-use efficiency than H. albiflorum but did not differ in efficiency of N use. H. radicata biomass increased faster in response to an N pulse than did the native H. albiflorum, as did the pool of N held in H. radicata tissues. • CONCLUSIONS: Our findings contrast with results from Hawaiian volcanic sites, where higher short-term resource use efficiency was reported for invasive species, including H. radicata. Our results suggest that at MSH, the success of H. radicata relies on rapid uptake and utilization of N rather than on higher efficiency. This strategy is especially advantageous at MSH because N pulses commonly occur as a consequence of herbivore-induced mortality of Lupinus lepidus (Fabaceae).

N-P co-limitation of primary production and response of arthropods to N and P in early primary succession on Mount St. Helens volcano.

BACKGROUND: The effect of low nutrient availability on plant-consumer interactions during early succession is poorly understood. The low productivity and complexity of primary successional communities are expected to limit diversity and abundance of arthropods, but few studies have examined arthropod responses to enhanced nutrient supply in this context. We investigated the effects of nitrogen (N) and phosphorus (P) addition on plant productivity and arthropod abundance on 24-yr-old soils at Mount St. Helens volcano. METHODOLOGY/PRINCIPAL FINDINGS: We measured the relative abundance of eight arthropod orders and five families in plots that received N, P, or no nutrients for 3-5 years. We also measured plant % cover, leaf %N, and plant diversity. Vegetation responded rapidly to N addition but showed a lagged response to P that, combined with evidence of increased N fixation, suggested P-limitation to N availability. After 3 yrs of fertilization, orthopterans (primarily Anabrus simplex (Tettigoniidae) and Melanoplus spp (Acrididae)) showed a striking attraction to P addition plots, while no other taxa responded to fertilization. After 5 yrs of fertilization, orthopteran density in the same plots increased 80%-130% with P addition and 40% with N. Using structural equation modeling, we show that in year 3 orthopteran abundance was associated with a P-mediated increase in plant cover (or correlated increases in resource quality), whereas in year 5 orthopteran density was not related to cover, diversity or plant %N, but rather to unmeasured effects of P, such as its influence on other aspects of resource quality. CONCLUSIONS/SIGNIFICANCE: The marked surprising response to P by orthopterans, combined with a previous observation of P-limitation in lepidopteran herbivores at these sites, suggests that P-mediated effects of food quantity or quality are critical to insect herbivores in this N-P co-limited primary successional system. Our results also support a previous suggestion that the availability of N in these soils is P-limited.

Successional change in phosphorus stoichiometry explains the inverse relationship between herbivory and lupin density on Mount St. Helens.

BACKGROUND: The average nitrogen-to-phosphorus ratio (NratioP) of insect herbivores is less than that of leaves, suggesting that P may mediate plant-insect interactions more often than appreciated. We investigated whether succession-related heterogeneity in N and P stoichiometry influences herbivore performance on N-fixing lupin (Lupinus lepidus) colonizing primary successional volcanic surfaces, where the abundances of several specialist lepidopteran herbivores are inversely related to lupin density and are known to alter lupin colonization dynamics. We examined larval performance in response to leaf nutritional characteristics using gelechiid and pyralid leaf-tiers, and a noctuid leaf-cutter. METHODOLOGY/PRINCIPAL FINDINGS: We conducted four studies. First, growth of larvae raised on wild-collected leaves responded positively to leaf %P and negatively to leaf carbon (%C), but there was no effect of %N or quinolizidine alkaloids (QAs). Noctuid survival was also positively related to %P. Second, we raised gelechiid larvae on greenhouse-grown lupins with factorial manipulation of competitors and soil N and P. In the presence of competition, larval mass was highest at intermediate leaf NratioP and high %P. Third, survival of gelechiid larvae placed on lupins in high-density patches was greater when plant competitors were removed than on controls. Fourth, surveys of field-collected leaves in 2000, 2002, and 2003 indicated that both %P and %N were generally greater in plants from low-density areas. QAs in plants from low-density areas were equal to or higher than QAs in high-density areas. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that declines in lupin P content under competitive conditions are associated with decreased larval growth and survival sufficient to cause the observed negative relationship between herbivore abundance and host density. The results support the theoretical finding that declines in stoichiometric resource quality (caused here by succession) have the potential to cause a decrease in consumer abundance despite very dense quantities of the resource.

Linking community and ecosystem development on Mount St. Helens.

In the two decades following the 1980 eruption of Mount St. Helens in Washington State, the N2-fixing colonizer Lupinus lepidus is associated with striking heterogeneity in plant community and soil development. We report on differences in nutrient availability and plant tissue chemistry between older, dense patches (core) of L. lepidus and more recently established low density patches (edge). In addition, we conducted a factorial nitrogen and phosphorus fertilization experiment in core patches to examine the degree of N and P limitation in early primary succession. We found that there were no significant differences in N or P availability between core and edge L. lepidus patches during the dry summer months, although nutrient availability is very low across the landscape. In the high density patches we found lower tissue N content and higher fiber content in L. lepidus tissue than in the younger edge patches. The addition of nutrients substantially altered plant community composition, with N addition causing an increase in other forb biomass and a corresponding competition-induced decline in L. lepidus biomass. The majority of the positive biomass response came from Hypochaeris radicata. In the second year of the fertilization experiment, the addition of N significantly increased total community biomass while L. lepidus biomass declined by more than 50%. The response of every species other than L. lepidus to N additions suggests that N may be the macronutrient most limiting plant production on Mount St. Helens but that the gains in productivity were somewhat offset by a decline of the dominant species. By the third year of the experiment, L. lepidus began to increase in abundance with P addition. This result suggests co-limitation of the community by N and P.

Partial shotgun sequencing of the Boechera stricta genome reveals extensive microsynteny and promoter conservation with Arabidopsis.

Comparative genomics provides insight into the evolutionary dynamics that shape discrete sequences as well as whole genomes. To advance comparative genomics within the Brassicaceae, we have end sequenced 23,136 medium-sized insert clones from Boechera stricta, a wild relative of Arabidopsis (Arabidopsis thaliana). A significant proportion of these sequences, 18,797, are nonredundant and display highly significant similarity (BLASTn e-value < or = 10(-30)) to low copy number Arabidopsis genomic regions, including more than 9,000 annotated coding sequences. We have used this dataset to identify orthologous gene pairs in the two species and to perform a global comparison of DNA regions 5' to annotated coding regions. On average, the 500 nucleotides upstream to coding sequences display 71.4% identity between the two species. In a similar analysis, 61.4% identity was observed between 5' noncoding sequences of Brassica oleracea and Arabidopsis, indicating that regulatory regions are not as diverged among these lineages as previously anticipated. By mapping the B. stricta end sequences onto the Arabidopsis genome, we have identified nearly 2,000 conserved blocks of microsynteny (bracketing 26% of the Arabidopsis genome). A comparison of fully sequenced B. stricta inserts to their homologous Arabidopsis genomic regions indicates that indel polymorphisms >5 kb contribute substantially to the genome size difference observed between the two species. Further, we demonstrate that microsynteny inferred from end-sequence data can be applied to the rapid identification and cloning of genomic regions of interest from nonmodel species. These results suggest that among diploid relatives of Arabidopsis, small- to medium-scale shotgun sequencing approaches can provide rapid and cost-effective benefits to evolutionary and/or functional comparative genomic frameworks.

Directed mutagenesis confirms the functional importance of positively selected sites in polygalacturonase inhibitor protein.

Polygalacturonase inhibitor proteins (PGIPs) protect plants against invasion by diverse microbial and invertebrate enemies that use polygalacturonase (PG) to breach the plant cell wall. Directed mutagenesis has identified specific natural mutations conferring novel defensive capability in green bean PGIP against a specific fungal PG. These same sites are identified as positively selected by phylogenetic codon-substitution models, demonstrating the utility of such models for connecting retrospective comparative analyses with contemporary, ecologically relevant variation.