Incorporating differences between genetic diversity of trees and herbaceous plants in conservation strategies

Reviews that summarize the genetic diversity of plant species in relation to their life history and ecological traits show that forest trees have more genetic diversity at population and species levels than annuals or herbaceous perennials. In addition, among-population genetic differentiation is significantly lower in trees than in most herbaceous perennials and annuals. Possible reasons for these differences between trees and herbaceous perennials and annuals have not been discussed critically. Several traits, such as high rates of outcrossing, long-distance pollen and seed dispersal, large effective population sizes (N_e), arborescent stature, low population density, longevity, overlapping generations, and occurrence in late successional communities, may make trees less sensitive to genetic bottlenecks and more resistant to habitat fragmentation or climate change. We recommend that guidelines for genetic conservation strategies be designed differently for tree species versus other types of plant species. Because most tree species fit an LH scenario (low [L] genetic differentiation and high [H] genetic diversity), tree seeds could be sourced from a few populations distributed across the species’ range. For the in situ conservation of trees, translocation is a viable option to increase N_e. In contrast, rare herbaceous understory species are frequently HL (high differentiation and low diversity) species. Under the HL scenario, seeds should be taken from many populations with high genetic diversity. In situ conservation efforts for herbaceous plants should focus on protecting habitats because the typically small populations of these species are vulnerable to the loss of genetic diversity. The robust allozyme genetic diversity databases could be used to develop conservation strategies for species lacking genetic information. As a case study of reforestation with several tree species in denuded areas on the Korean Peninsula, we recommend the selection of local genotypes as suitable sources to prevent adverse effects and to insure the successful restoration in the long term.     

Determination of trace perchlorate in water: a simplified method for the identification of potential interferences

Background

Perchlorate contamination of water and food poses potential health risks to humans due to the possible interference of perchlorate with the iodide uptake into the thyroid gland. Perchlorate has been found in food and drinking, surface, or swimming pool waters in many countries, including the United States, Canada, France, Germany, and Switzerland, with ion chromatography (IC) being the preferred analytical method. The standardization of a robust ion chromatographic method is therefore of the high interest for public health and safety. This article summarizes the experiments and results obtained from analyzing untreated samples, considering the sample’s electrical conductance as guidance for direct sample injection as described in EPA 314.0.

Results

The suitability of ion chromatography with suppressed conductivity detection was tested for water samples in order to check the influence of matrix effects on the perchlorate signal of untreated samples. A sample injection volume of 750 μL was applied to the selected 2 mm?IC?column. The IC?determination of perchlorate at low µg/L levels is challenged by the presence of high loads of matrix ions (e.g., chloride, nitrate, carbonate, and sulfate at 100 mg/L and above). Perchlorate recovery is impaired with the increasing matrix ion concentrations, and its chromatographic peak is asymmetric particularly at low perchlorate concentrations. The identification of the individual maximum concentration of interfering anions like chloride, nitrate, and sulfate that influence perchlorate recovery helps to reduce the number of sample preparation steps or an obligatory measurement of the electrical conductivity of the sample. Within the scope of this study, samples containing less than 125 mg/L of either anion did not need sample preparation.

Conclusion

The identification of the maximum concentration of interfering anions like chloride, nitrate, and sulfate influencing perchlorate recovery provides a simplified alternative to the EPA 314.0 method. This approach reduces unnecessary sample preparation steps while allowing a reliable prognosis of possible interferences and maintaining result quality. This study was performed to support the development of a respective international standard, which is being established by the International Organization for Standardization (ISO). The results of the study are also intended to be used as guidance for interested laboratories to optimize the analytical workflow for trace perchlorate determination.

     

Lead uptake by Eichhornia Crassipes

Factors influencing uptake of lead(II) by the water hyacinth (Eichhornia Crassipes), were examined. Two phases of uptake were observed for the concentration range investigated (0.01–1000 ppm). The initial, rapid uptake phase of about 4 hours is followed by a slower, near linear phase extending past 24 hours. Stirring the solution enhanced uptake, suggesting that lead removal is in part diffusion limited. In the range of 4–8, pH has little effect on uptake where as outside this range, uptake is reduced. Increased solution volume or rootmass results in more metal being removed by the plant. The presence of strong complexers blocks the initial rapid uptake phase as does the presence of Zn(II), Cd(II), Hg(II), and Fe(III). Strong complexers can also strip a portion of any lead already removed from solution by the plant.     

Effects of Widespread Drought‐Induced Aspen Mortality on Understory Plants

Forest die‐off around the world is expected to increase in coming decades as temperature increases due to climate change. Forest die‐off will likely affect understory plant communities, which have substantial influence on regional biological diversity, ecosystem function, and land–atmosphere interactions, but how die‐off alters these plant communities is largely unknown. We examined changes in understory plant communities following a widespread, drought‐induced die‐off of trembling aspen (Populus tremuloides) in the western United States. We assessed shrub and herbaceous cover and volume in quadrats in 55 plots located across a wide range of levels of aspen mortality. We measured species richness and composition of herbaceous plant communities by recording species presence and absence in 12 sets of paired (1 healthy, 1 dying) aspen plots. Although understory composition in healthy and dying stands was heterogeneous across the landscape, shrub abundance, cover, and volume were higher and abundance of herbaceous species, cover, and volume were lower in dying aspen stands. Shrub cover and volume increased from 2009 to 2011 in dying stands, which suggests that shrub growth and expansion is ongoing. Species richness of herbs declined by 23% in dying stands. Composition of herbs differed significantly between dying and healthy stands. Richness of non‐native species did not differ between stand types. The understory community in dying aspen stands was not similar to other shrub‐dominated plant communities in the region and may constitute a novel community. Our results suggest that changes in understory plant communities as forests die off could be a significant indirect effect of climate change on biological diversity and forest communities. Efectos de la Mortalidad Extensiva de Álamos Inducida por Sequía sobre Plantas del Sotobosque     

Mother’s choice of the oviposition site: balancing risk of egg parasitism and need of food supply for the progeny with an infochemical shelter?

Summary. Oviposition site selection of herbivorous insects depends primarily on host plant presence which is essential for offspring survival. However, parasitoids can exploit host plant cues for host location. In this study, we hypothesised that herbivores can solve this dilemma by ovipositing within high plant diversity. A diverse plant species composition might represent an ‘infochemical shelter’, as a potentially complex volatile blend can negatively affect the host location ability of parasitoids. We examined this exemplarily for the egg-laying response of the generalist leaf beetle, Galeruca tanaceti, in relation to (1) host plant availability and (2) plant species diversity in the field. Further, we investigated the effect of odours from mixed plant species compositions on (3) leaf beetle oviposition site selection and on (4) the orientation of its specialised egg parasitoid, Oomyzus galerucivorus. In the field, egg clutch occurrence was positively related to the presence and quantity of two major host plants, Achillea millefolium (yarrow) and Centaurea jacea, and to the number of herbaceous plant species. In two-choice bioassays, female beetles oviposited more frequently on sites surrounded by an odour blend from a diverse plant species composition (including yarrow) than on sites with a pure grass odour blend. In the presence of yarrow odour and an odour blend from a diverse plant mixture (including yarrow) no difference in the oviposition response was recorded. Experienced parasitoid females were attracted to yarrow odours, but showed no response when yarrow odours were offered simultaneously with odours of a non-host plant. In conclusion, it could be shown in laboratory bioassays that the parasitoid responds only to pure host plant odours but not to complex odour blends. In contrast, the herbivore prefers to oviposit within diverse vegetation in the field and in the laboratory. However, the laboratory results also point to a priority of host plant availability over the selection of a potential ‘infochemical shelter’ for oviposition due to high plant diversity.     

The role of archaeophytes and neophytes in the Dutch coastal dunes

The position of alien plant species in the Dutch coastal dune vegetation is evaluated considering 12 archaeophytes and 20 neophytes (including one moss), all of widespread occurrence in the coastal area of the Netherlands. Almost all archaeophytes have become part of natural vegetation types. Open scrub communities, in particular Hippophae rhamnoides-Sambucus nigra scrub at the leeside of the outer dunes, and calciphilous moss-dominated pioneer vegetation are relatively rich in archaeophytes. Among neophyte vascular species a contrast is apparent between herbaceous and woody species. The majority of herbaceous neophytes are characteristic of man-disturbed habitats and are only infrequently observed in natural vegetation types. However, woody species (as well as the moss Campylopus introflexus) have entered into natural vegetation on a large scale and behave in an invasive way.     

Heterotrophic nutrition of the marine pennate diatom Nitzschia angularis var. affinis

The nutritional pattern for heterotrophic growth of Nitzschia angularis var. affinis (Grun.) Perag. is more complex than for other diatom species studied previously. This species grew slowly in the dark in the presence of single amino acids, either glutamate or alanine; other amino acids when supplied singly were not used as substrates. Carbon from glutamate was converted to cell carbon with an efficiency of 43%. Glutamine was inhibitory both in the light and in the dark, and aspartate inhibited heterotrophic growth on glutamate. Glucose and tryptone supplied singly did not support heterotrophic growth, but when combined, together they allowed for rapid growth of N. angularis (generation time of 16 h). Glucose in combination with glutamate, alanine, aspartate, or asparagine (but not with any other amino acids) also supported growth in the dark, at a rate considerably more rapid than with glutamate alone. In the presence of excess glucose and limiting concentrations of glutamate, approximately 50% of the cell carbon for heterotrophic growth came from glucose, while in combination with tryptone about 25% of the cell carbon came from glucose. Amino acids were taken up by cells grown either photoautrophically or in the dark in the presence or absence of organic substrates; uptake rates were some-what higher for dark-grown than for light-grown cells. Glucose was taken up only by dark-grown cells; induction of a glucose uptake system in the dark required the presence of glutamate but not of glucose. The rates of uptake of glutamate and glucose by cells incubated in the dark with glutamate were sufficiently high to account for the observed rates of growth on these substrates in the dark. The uptake systems of N. angularis have relatively high affinities for glucose (K _s =0.03 mM) and glutamate (K _s =0.02 mM).Contribution No. 890 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Phytoaccumulation of iron by callus tissue of Clerodendrum indicum (L)

Clerodendrum indicum (L.) is one of the important medicinal plants, originating in India. The plant is used as medicine for bronchitis, asthma and different immunological disease. Micro-propagation is a quick method, producing a huge number of plants. Apical leaf of Clerodendrum indicum (L.) was used as an explant for callogenesis. The combinations of different plant growth regulators (2,4-D, NAA, KN) in MS (Murashige and Skoog) medium produced two types of calli: one was friable and loose and the other was green, embryogenic and compact. Heavy metals cause pollution and health hazards. The plant absorbs heavy metals, which is very useful for controlling environmental pollution. Iron is a major nutrient for both plants and animals, but hyper accumulation of iron is injurious to health. Calli were used to study the optimum conditions of iron uptake in presence of different chemical environments. The different environments were created by using different chemical reagents such as (HCL, HNO₃, H₃PO₄ and KNO₃). Iron (III) was spiked with radioactive Fe-59 and dynamics of uptake was followed by measuring radioactivity by gamma-ray spectrometer. The best condition of iron uptake corresponded to 1 mol L^?1 KNO₃ after 4 hour of equilibration.     

Arbuscular mycorrhiza confers lead tolerance and uptake in Miscanthus sacchariflorus

The effects of an arbuscular mycorrhizal fungi (AMF) association on the growth, survival capabilities, nutrients and lead (Pb) uptake of Miscanthus sacchariflorus under different Pb concentrations were studied in the form of pot cultures. The treatments comprised inoculation or non-inoculation of the AMF, Gigaspora margarita, and the addition of three Pb concentrations to the soil (0, 100 and 1000?mg?kg^?1). The addition of Pb significantly decreased mycorrhizal colonisation. The inoculation of AMF with Pb increased chlorophyll content, Fv/Fm, total dry mass, indole-3-acetic acid (IAA), total nitrogen, and total phosphorus, whereas H₂O₂ level, indole-3-acetic acid oxidase (IAAO) activity, and peroxidase (POD) activity were low compared to those in the non-inoculated treatments. Moreover, the application of AMF together with Pb doses induces concentrations of Pb in the plant, where the higher dose of Pb (1000?mg?kg^?1) induces a lower content of Pb in the aerial part of the plant but a higher content in the root. G. margarita enhanced the tolerance of M. sacchariflorus against Pb toxicity, and facilitated the accumulation of Pb in the plant roots, whereas translocation to the shoots was inhibited at the highest dose Pb (1000?mg?kg^?1). However, in contaminated soil, the Pb removal capability of M. sacchariflorus with AMF was remarkable.     

砷浓度、形态及碳酸氢盐对蜈蚣草吸收砷的影响

为了探讨超富集植物蜈蚣草在处理高砷地下水方面的可行性,研究了水培条件下砷的浓度、形态和碳酸氢盐(HCO-3)对超富集植物蜈蚣草吸收砷的影响。实验中使用了浓度为0.1～100mg·L-1的As(III)和As(V)溶液。HCO-3处理中,HCO-3浓度范围为0.5～20mmol·L-1,As(III)或As(V)的浓度为5mg·L-1。结果表明,在水培条件下,蜈蚣草具有明显的耐高砷特征。当介质砷含量高达100mg·L-1时,砷的去除率可达到80%,且对As(III)的吸收效率高于As(V)。植物体内砷形态研究表明,蜈蚣草体内2种形态砷的含量与外源砷形态有一定的关系,As(V)处理条件下,植物体中的As(V)比例较As(III)处理高。高浓度的HCO-3(20mmol·L-1)处理对蜈蚣草地上部分生物量没有明显影响,但是抑制了地下部分的生长,并且对砷的吸收表现出明显的抑制作用。     

Seasonal Variations in the Metal Concentration of Taraxacum Officinale,Plantago Major and Plantago Lanceolata

The seasonal changes in the concentrations of aluminium (Al), calcium (Ca), cadmium (Cd), chromium (Cr), copper (Cu), magnesium (Mg), manganese (Mn), lead (Pb), and zinc (Zn) in the leaves of Taraxacum officianale, Plantago lanceolata and Plantago major are investigated. the most convenient periods for sampling are established when the elemental concentrations have minimal variation and are independent on the development of the plant itself. On this basis, the presumption is made that the most appropriate sampling period for herbaceous species for biomonitoring processes is the end of blossoming of the plant or immediately after that.     

砷浓度、形态及碳酸氢盐对蜈蚣草吸收砷的影响

大同盆地是典型的高砷地下水分布区。利用从地方性砷中毒严重病区山阴县采集的高砷地下水样品,用稀释培养法实验研究了外加砷源对地下水中微生物数量的影响;同时基于生物学可培养法和16S rDNA序列比对法,选取代表性高砷水样,研究了耐砷菌的种群特征。结果表明,外加砷源对地下水中微生物数量影响显著,高浓度砷会抑制大部分微生物生长,使微生物数量减少;低浓度砷对微生物生长具有一定促进作用。通过多次分离、纯化从3个不同砷含量地下水样中分离到多株砷抗性菌,经鉴定属于主要为Bacillus、Pseudomonas、Paenibacillus、Aeromonas、Enterobacter5个属。从RDP(Ribosomal Database Project)分析显示3个水样可培养微生物组成不同,都有生存能力强能够耐低浓度NaAsO₂的Bacillales,优势耐砷菌是γ-proteobacteria,其中Enterbacter具有耐高浓度NaAsO₂的能力。     

Study of metal accumulation in wild plants using principal component analysis and ionic impulsions

The objective of this work is to apply principal component analysis (PCA) and ionic impulsions to the study of the accumulation and uptake of metals in wild plants. Soil samples were taken in three locations and the following plant species were selected: Poa, Pteridium aquilinum, Diplotaxis, Plantago lanceolata and Trifolium repens. After determining contents of Na, K, Ca, Mg, Mn, Pb, Co, Ni, Cu, Cr, Zn, Cd and Fe in the plant samples and in the soils, principal component analysis was carried out for plant data treatment. Sample position maps revealed that there was no substantial differences among samples when plant species or sampling locations were used as a criteria. As the factors obtained by PCA only correlated with one or two variables, the study of accumulation patterns of metals was carried out by using ionic impulsions. The presence of large amounts of pollutants (like Cd, Cr) in plants is related to a greater assimilation of essential micronutrients in order to counteract the influence of the pollutants.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. IV. Transient response of Chaetoceros debilis,Skeletonema costatum,and Thalassiosira gravida to a single addition of the limiting nutrient

The kinetic response of ammonium- or silicate-limited and ammonium- or silicatestarved populations of Chaetoceros debilis, Skeletonema costatum, and Thalassiosira gravida was determined by a single addition of the limiting nutrient to a steady-state culture and subsequent monitoring of the nutrient disappearance of the limiting and non-limiting nutrients at frequent time intervals. The kinetic response of nonlimited (nutrient) populations of these three species was also determined. Three distinct modes of the uptake of the limiting nutrient were observed for ammonium-or silicate-limited populations of these three species, surge uptake (V _s ), internally (cellular) controlled uptake (V _i ), and externally (ambient limiting nutrient concentration) controlled uptake (V _e ). Non-limited populations did not exhibit the three distinct segments of uptake, V _s , V _i and V _e . Estimates of the maximal uptake rate (V _max) and the Michaelis constant (K _s ) were obtained from nutrient-limited populations during the V _e segment of the uptake curve. Pooled values of V _e for the three ammonium-limited populations yielded V _max and K _s estimates of 0.16 h^-1 and 0.5 g-at NH₄–N l^-1. Kinetic data derived from the V _e segment of the uptake curve for silicate-limited populations yielded different values of V _max and K _s for each of the three species. In a number of parameters that were measured, T. gravida was clearly different from C. debilis and S. costatum and its recovery from nutrient starvation was the slowest. Recovery of all species from silicate limitation or starvation was slower than from ammonium limitation or starvation. Ammonium-starved populations maintained a maximal uptake rate at a substrate concentration an order of magnitude lower (0.1 g-at NH₄–N l^-1) than that observed for NH₄-limited populations (1.0 g-at NH₄–N l^-1). Adaptation to the severity of the nutrient limitation occurred as changes in the magnitude of cellular characteristics, such as short-term uptake potential (V _s ) and affinity for the substrate (K _s ). The consequence of these results are discussed in terms of another possible mechanism to explain changes in species composition and succession in nutrient-depleted environments.Contribution No. 944 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Effectiveness of emergent and submergent aquatic plants in mitigating a nitrogen–permethrin mixture

Hydraulically connected wetland microcosms (～50?L) in series were used to test the effectiveness of varying combinations of two common aquatic vascular plants, parrot feather (Myriophyllum aquaticum) and cattail (Typha latifolia), for mitigating contamination from a mixture of nitrogen (ammonium nitrate) and permethrin. The upstream series included Myriophyllum only (M) and Typha only (T) while the combination upstream effluent into downstream series included Myriophyllum into Myriophyllum (M?+?M) and Typha into Myriophyllum (T?+?M). During flow, M into M?+?M more efficiently mitigated nitrogen than T into T?+?M. Post-flow, nitrogen removal efficiency was greater for T versus M and M?+?M versus T?+?M. Mean aqueous dissipation half-lives (t_1/2) of NH₄-N and NO₃-N were more rapid in T than M treatments. Ammonium and nitrate t_1/2 was highly correlated with aquatic plant above-ground shoot biomass. Permethrin mitigation efficiencies and t_1/2 were not significantly affected by plant species either singly or in combination. Trans-permethrin t_1/2 was moderately correlated with plant biomass, but not cis-permethrin t_1/2. Results of this study indicate differences in aquatic plant species and flow path influence nitrogen removal but not permethrin. However, plant species appears less important than overall plant biomass in ascertaining aquatic plant effectiveness in mitigating a nitrogen–permethrin mixture.     

Accumulation, distribution and transformation of DDT and PCBs by Phragmites australis and Oryza sativa L.: II. Enzyme study

Two wetland plant species, Phragmites australis and Oryza sativa, were grown in a glasshouse under hydroponics conditions. Enzyme extracts from different parts of the plants were used to determine the transformation rate of o,p′-DDT, p,p′-DDT and PCBs. The organic pollutants were directly spiked into the enzyme extracts, and samples were collected every 30 min and analyzed with a GC-ECD. Root extracts of P. australis readily degraded and transformed DDT and some PCB congeners with a low degree of chlorination. In contrast, crude extracts of O. sativa showed no appreciable degradation or transformation of DDT or PCBs. Inhibition studies indicated that the degradation and transformation of both DDT and PCBs by P. australis enzymes were partly mediated by peroxidase and the plant P-450 system. PCBs with a high degree of chlorination were highly resistant to transformation or degradation by plant enzymes. Both wetland plant species accumulated substantial quantities of the persistent organic chemicals but had different degradation capacities. The enzyme systems in P. australis were much more effective that those in rice in the degradation and transformation of the organic pollutants.     

Antimony uptake by <Emphasis Type="Italic">Zea mays</Emphasis> (L.) and <Emphasis Type="Italic">Helianthus annuus</Emphasis> (L.) from nutrient solution

We investigated the extent of Sb uptake by maize (Zea mays) and sunflower (Helianthus annuus) from nutrient solutions containing concentrations from 3 to 24 mg/L of potassium antimonate, with the aim of determining the potential of Sb to enter the food chain. The maximum shoot Sb concentrations in Z. mays and H. annuus were 41 mg/kg and 77 mg/kg dry weight, respectively. There was no significant difference in Sb uptake between species. The average bioaccumulation coefficients (the plant/solution concentration quotients) were 1.02 and 1.93 for Z. mays and H. annuus, respectively. Phosphate addition did not affect plant growth or Sb uptake. Antimony uptake by both Z. mays and H. annuus is unlikely to pose a health risk to animals and humans.     

Comparisons of nutrient uptake rates for Baltic macroalgae with different thallus morphologies

In-situ experiments were performed during different seasons to determine uptake rates of PO _3- ⁴ , NH ₊ ⁴ and NO _- ³ within ecologically representative ranges of nutrient concentrations, of dominant macroalgae in the Baltic Sea. Uptake rates were governed by nutrient concentrations, water temperature and thallus morphology, but not by the phylogenetic affinity of the species. Nitrogen uptake rates were always higher than those of phosphorus at the same concentrations, and NH ₊ ⁴ –N uptake rates exceeded those of NO _- ³ –N. The lowest uptake rates occurred among the late successional, long-lived, coarse species with low surface: volume ratios (Fucus vesiculosus, Furcellaria lumbricalis andPhyllophora truncata). The highest uptake rates were measured for short-lived, opportunistic algae, filamentous or with numerous hairs, (Cladophora glomerata, Enteromorpha ahlneriana, Scytosiphon lomentaria, Dictyosiphon foeniculaceus andCeramium tenuicorne). The latter group also had the highest V_max:k_max ratios, which indicates a more competitive advantage for nutrient uptake at low concentrations.     

Uptake and elimination kinetics of triphenyltin hydroxide by two fish species

The uptake from water and the elimination of carbon‐14 radiolabelled triphenyltin hydroxide ([¹⁴C]TPTH) was studied in two fish species: guppy (Poecilia reticulata) and rainbow trout larvae (Salmo gairdneri). During all the experiments no steady state in fish was found. TPTH was rapidly taken up, while elimination was very slow. Uptake and elimination rate constants (k ₁ and k ₂, respectively), and a bioconcentration factor were estimated, assuming first order kinetics.

During eight days of exposure of guppy to TPTH an uptake rate constant k ₁ of 70±7L/kg.d and an elimination rate constant k ₂ of 0.005 ±0.029d ^‐1 was found. This resulted in a biconcentration factor of at least 2.1 × 10³L/kg (wet weight). Comparable results were obtained during a 30 days exposure experiment with guppy: k ₁ was 41±2L/kg.d, k ₂ 0.014 + 0.002d^‐1, and the bioconcentration factor was estimated to be 2.9 × 10³ L/kg (wet weight).

Four days exposure of rainbow trout larvae resulted in a it, of 22+ 2 L/kg.d, and a k ₂ of 0.031 ±0.007d^‐1. Using these k, and k ₂ values it was estimated that the biconcentration factor exceeds 650 L/kg (wet weight).     

Nonadditive effects among threats on rare plant species

The current loss of biodiversity has put 50,000 plant species at an elevated risk of extinction worldwide. Conserving at-risk species is often complicated by covariance or nonadditivity among threats, which makes it difficult to determine optimal management strategies. We sought to demographically quantify covariance and nonadditive effects of more threats on more rare plant species than ever attempted in a single analysis. We used 1082 population reports from 186 populations across 3 U.S. states of 27 rare, herbaceous plant species collected over 15 years by citizen scientists. We used a linear mixed-effects model with 4 threats and their interactions as fixed predictors, species as a random predictor, and annual growth rates as the response. We found a significant 3-way interaction on annual growth rates; rare plant population sizes were reduced by 46% during the time immediately after disturbance when populations were also browsed by deer (Odocoileus virginianus) and had high levels of encroachment by woody species. This nonadditive effect should be considered a major threat to the persistence of rare plant species. Our results highlight the need for comprehensive, multithreat assessments to determine optimal conservation actions.