Conservation genetic inferences in the carnivorous pitcher plant <Emphasis Type=Italic>Sarracenia alata</Emphasis> (Sarraceniaceae)

Conservation geneticists make inferences about their focal species from genetic data, and then use these inferences to inform conservation decisions. Since different biological processes can produce similar patterns of genetic diversity, we advocate an approach to data analysis that considers the full range of evolutionary forces and attempts to evaluate their relative contributions in an objective manner. Here we collect data from microsatellites and chloroplast loci and use these data to explore models of historical demography in the carnivorous Pitcher Plant, Sarracenia alata. Findings indicate that populations of S. alata exhibit high degrees of population genetic structure, likely caused by dispersal limitation, and that population sizes have decreased in western populations and increased in eastern populations. These results provide new insight to the management and conservation of plants restricted to small, declining populations isolated in increasingly scarce and highly threatened habitat, including other rare and endangered species of Sarracenia.     

Efficiency of insect capture by Sarracenia purpurea (Sarraceniaceae), the northern pitcher plant

Pitcher plants (Sarracenia purpurea L.) attract insects to pitchers and then capture them in fluid-filled, pitfall traps, but how efficient are pitcher plants at capturing prey in their natural environment? We monitored insect activity by videotaping pitchers and analyzing videotapes for several variables including identity of each visitor and outcome of each visit (e.g., departure or capture). Efficiency of capture (i.e., number of captures per number of visits) was low. Overall efficiency of capture was 0.83–0.93%, depending on whether potential prey were broadly or narrowly defined. Ants constituted 74% of the potential prey. Efficiency of capture of ants was even lower at 0.37%. Potential prey were more likely to visit pitchers with greater red venation and less water in the pitcher. There was no correlation between number of potential prey visiting a pitcher and pitcher age, length, or mouth width. Also, number of potential prey visits did not correlate with plant size, air temperature, time of day or date of videotaping. While the overall efficiency of prey capture was very low, pitcher plants may still benefit from the additional nutrients. However, the relationship between ants and S. purpurea remains an enigma, since it is unclear whether the plants capture enough ants to compensate for nectar lost to ants.     

Limits to reproductive success of Sarracenia purpurea (Sarraceniaceae)

Plant biologists have an enduring interest in assessing components of plant fitness and determining limits to seed set. Consequently, the relative contributions of resource and pollinator availability have been documented for a large number of plant species. We experimentally examined the roles of resource and pollen availability on seed set by the northern pitcher plant Sarracenia purpurea. We were able to distinguish the relative contributions of carbon (photosynthate) and mineral nutrients (nitrogen) to reproductive success. We also determined potential pollinators of this species. The bees Bombus affinis and Augochlorella aurata and the fly Fletcherimyia fletcheri were the only floral visitors to S. purpurea that collected pollen. Supplemental pollination increased seed set by <10%, a much lower percentage than would be expected, given data from noncarnivorous, animal-pollinated taxa. Seed set was reduced by 14% in plants that could not capture prey and by another 23% in plants whose pitcher-shaped leaves were also prevented from photosynthesizing. We conclude that resources are more important than pollen availability in determining seed set by this pitcher plant and that reproductive output may be another "cost" of the carnivorous habit.     

Seed dispersal and seedling establishment of Sarracenia purpurea (Sarraceniaceae)

Plant ecologists continue to grapple with Reid's paradox, the observation that dispersal distances of most herbs and trees are too limited to account for their recolonization of northern latitudes following glacial recession. As global climate changes and natural habitats become increasingly fragmented, understanding patterns of seed dispersal and the potential for long-distance colonization takes on new importance. We studied the dispersal and establishment of the northern pitcher plant Sarracenia purpurea, which grows commonly in isolated bogs throughout Canada and eastern North America. Median dispersal distance of S. purpurea is only 5 cm, which is insufficient to explain its occurrence throughout formerly glaciated regions of North America. Establishment probability of seeds in the field is approximately 5%, and juveniles are normally found clustered around adult plants. The large-scale population genetic structure of this species can be accounted for by rare long-distance dispersal events, but its predictable occurrence in isolated habitats requires additional explanation. Reid's paradox remains an open question, and predicting long-range colonization into fragmented habitats by species with limited dispersal ability is a novel challenge.     

Inbreeding, outbreeding, and heterosis in the yellow pitcher plant, Sarracenia flava (Sarraceniaceae), in Virginia

The yellow pitcher plant, Sarracenia flava, is an insectivorous plant restricted to fire-maintained wetland ecosystems in southeastern Virginia. Only four natural sites remain in the state totaling fewer than 100 clumps. Plants from sites located in Dinwiddie, Greensville, Prince George, Sussex counties, and the city of Suffolk were tested for the effects of self-pollination, intrasite outcrossing, and intersite outcrossing on offspring quantity (total seed number and total seed mass) and offspring quality (avarage seed mass, germination, and growth).Self-pollination resulted in significantly lower offspring quantity and quality. Total seed number and total seed mass for self-pollinated capsules were approximately one-fourth that of outcrossed capsules. Germination, survivorship, and growth over 5 yr were also significantly lower for offspring from self-pollinated capsules. Together, these results suggest strong inbreeding depression in this species.Relative to offspring from intrasite crosses, offspring from intersite crosses were significantly larger after 5 yr of growth. This suggests that restoration efforts for Virginia S. flava will be most successful when plants from multiple sites are used.     

Nutrient limitation and morphological plasticity of the carnivorous pitcher plant Sarracenia purpurea in contrasting wetland environments

* Plasticity of leaf nutrient content and morphology, and macronutrient limitation were examined in the northern pitcher plant, Sarracenia purpurea subsp. purpurea, in relation to soil nutrient availability in an open, neutral pH fen and a shady, acidic ombrotrophic bog, over 2 yr following reciprocal transplantation of S. purpurea between the wetlands. * In both wetlands, plants were limited by nitrogen (N) but not phosphorus (P) (N content < 2% DW(-1), N : P < 14) but photosynthetic quantum yields were high (F(V)/F(M) > 0.79). Despite carnivory, leaf N content correlated with dissolved N availability to plant roots (leaf N vs , r(2) = 0.344, P < 0.0001); carnivorous N acquisition did not apparently overcome N limitation. * Following transplantation, N content and leaf morphological traits changed in new leaves to become more similar to plants in the new environment, reflecting wetland nutrient availability. Changes in leaf morphology were faster when plants were transplanted from fen to bog than from bog to fen, possibly reflecting a more stressful environment in the bog. * Morphological plasticity observed in response to changes in nutrient supply to the roots in natural habitats complements previous observations of morphological changes with experimental nutrient addition to pitchers.     

Morphological variation in Sarracenia purpurea (Sarraceniaceae): geographic, environmental, and taxonomic correlates

Geographic variation in morphology reflects phenotypic responses to environmental gradients and evolutionary history of populations and species and may indicate local or regional changes in environmental conditions. The pitcher plant (Sarracenia purpurea) illustrates these principles. At local scales, its morphology reflects nutrient availability. At points along its broad geographic range (from Florida to northern Canada) morphology has been used to distinguish subspecies and varieties, but there has been no detailed study of the continuum of morphological variation across this entire range. Patterns of morphological variation in S. purpurea were characterized as a function of climatic and environmental conditions at 39 sites spanning its range. Differences in pitcher size and shape were strongly correlated with temperature, annual precipitation, and availability of ammonium and calcium in peat pore water. Pitcher shape (lip width, mouth diameter, and pitcher width) in Florida panhandle populations differed significantly from pitcher shape of all other populations, even after accounting for environmental correlations. In contrast, the northern and southern subspecies of S. purpurea (the latter exclusive of the Florida panhandle populations) cannot be distinguished based on these morphological measurements alone. These results support a recent proposal that identifies the Florida populations as a distinct species, Sarracenia rosea.     

Interspecific and intraspecific variation in seed size and germination requirements of Sarracenia (Sarraceniaceae)

Seed size and germination requirements of eight (of nine) Sarracenia species, and 13 populations of S. purpurea were studied. All species except for S. purpurea are restricted to the southeastern United States, whereas S. purpurea ranges across Canada, southward along the eastern United States into Maryland and Virginia (S. purpurea ssp. purpurea), and from New Jersey southward into northern Florida and the coast of the Gulf of Mexico (S. purpurea ssp. venosa). I tested the hypotheses that dormancy-breaking requirements vary predictably among species across a latitudinal gradient. I also sought to determine whether seed size and germination requirements were useful characters for resolving systematic and phylogenetic questions within this genus. Seed size varied significantly among species, but variability in seed size within S. purpurea exceeded the variability in seed size observed across all eight species studied. Seeds of all species are morphophysiologically dormant upon dispersal. Length of required cool, moist pretreatment varied among species, and germination in higher latitude populations is enhanced with longer pretreatment. In contrast, variability in germination requirements of subspecies, varieties, and populations of the geographically wide-ranging S. purpurea was not related clearly to geographic location (latitude or elevation). Germination requirements do not map onto a proposed phylogeny of Sarracenia, but observed differences in germination requirements of S. purpurea ssp. venosa var. burkii relative to other populations of S. purpurea support the recent proposal to elevate this variety to species status.     

Development of microalgae communities in the Phytotelmata of allochthonous populations of Sarracenia purpurea (Sarraceniaceae)

The phytotelmata of the North American pitcher plant Sarracenia purpurea are colonised by a great variety of aquatic organisms and, thus, provide an ideal model to study trophic interactions in small freshwater ecosystems. Although algae are discussed as a potential food source for predators, little is known about the structure of algae coenoses in pitchers of S. purpurea. This study aims to elucidate temporal shifts in the algae community structure in pitchers of an allochthonous population of S. purpurea in Saxony, Germany. A total of 78 algae taxa was found in the pitchers. Mean algae abundances in new and old pitchers were similar (2.6 x 10(5) and 2.3 x 10(5) algae ml(-1), respectively). Taxa from the orders Chlamydomonadales, Chlorococcales, and Ochromonadales were the primary colonisers. With increasing age of the pitchers the filamentous green algae from the order Klebsormidiales became more abundant. In contrast, pennate diatoms dominated the algae coenoses in the fen. Algae community structure in vase-shaped 50 ml Greiner tubes was similar to those of natural pitchers. Differences in the temporal patterns of algae coenoses in individual pitchers suggested a colonisation of the pitchers by algae via trapped insects, air and rain water rather than via the surrounding fen. Biomass of algae approximated 0.3 mg C ml(-1), which corresponds to 82.8 % of the living biomass (bacteria, heterotrophic nanoflagellates, algae, protozoans and rotifers). Rotifers were abundant in new pitchers; nematodes and mites were seldom found in all pitchers. A similar qualitative and quantitative composition of the aquatic biocoenoses was observed in pitchers of another allochthonous S. purpurea population growing in Blekinge, Sweden. Biomass of algae represented nearly one quarter of the total organic matter content in the pitchers. Thus, nitrogen and phosphorus compounds present in the algae biomass might be used by the carnivorous S. purpurea plant as additional food source in allochthonous populations in Europe lacking top predators.     

Fungal endophyte diversity in Sarracenia

Fungal endophytes were isolated from 4 species of the carnivorous pitcher plant genus Sarracenia: S. minor, S. oreophila, S. purpurea, and S. psittacina. Twelve taxa of fungi, 8 within the Ascomycota and 4 within the Basidiomycota, were identified based on PCR amplification and sequencing of the internal transcribed spacer sequences of nuclear ribosomal DNA (ITS rDNA) with taxonomic identity assigned using the NCBI nucleotide megablast search tool. Endophytes are known to produce a large number of metabolites, some of which may contribute to the protection and survival of the host. We speculate that endophyte-infected Sarracenia may benefit from their fungal associates by their influence on nutrient availability from within pitchers and, possibly, by directly influencing the biota within pitchers.     

Local genetic differentiation in Proteopsis argentea (Asteraceae), a perennial herb endemic in Brazil

 Proteopsis argentea (Asteraceae, Vernonieae) is a perennial herb endemic to the “campos rupestres” of the Espinha?o Range in Minas Gerais, Brazil, with fragmented populational distribution. Eleven populations were sampled, throughout the entire distribution of the species, and assayed for isozyme variation. Low intra-population genetic diversity was found (P = 19.2; A = 1.30; He = 0.058) whereas species level diversity was higher (P = 55.5, A = 2.0, He = 0.093). The most geographically isolated population showed exclusive alleles at two loci, whereas two populations less than 2 km apart from each other showed inverted frequencies for two alleles. Mean genetic identity was high (I = 0.974), but the large Fst (0.30) indicates that the species could lose an important part of its genetic variation with the extinction of a single population. Our findings indicate that geographic isolation alone cannot explain population differentiation: localized pollinator behaviour and selection, for example, may be contributing to the patterns observed. Received February 18, 2000 Accepted November 1, 2000     

Low levels of genetic variation inPhenakospermum guyannense (Strelitziaceae), a widespread bat-pollinated Amazonian herb

Phenakospermum guyannense is a monotypic, arborescent, long-lived monocot that is widespread in Amazonian South America. This outcrossing species is pollinated primarily by phyllostomid bats. Given these life-history characteristics,P. guyannense is expected to exhibit high levels of genetic variation and gene flow. We used isozyme electrophoresis and randomly amplified polymorphic DNA (RAPD) to characterize genetic variation in populations ofP. guyannense from French Guiana. Both measures detected a surprisingly low level of genetic variation, with only five out of twenty (25%) allozyme loci polymorphic (P), 1.35 alleles per locus (A), and an expected heterozygosity (H_e) of 0.090 at the species level. Isozymic genetic variation was even lower within populations (P = 17.5, A = 1.24, H_e = 0.074), and was corroborated by a RAPD assay that used 26 arbitrary primers (P = 3.61, A = 1.04, H_e = 0.014). Although overall levels of variation were low, the detectable variation was distributed as would be expected for an outcrossing species with extensive gene flow (mean G_ST = 0.230). We suspect thatP. guyannense is depauperate in genetic variation because of a series of bottlenecks that affected the species over this portion of its range.     

High genetic diversity vs. low genetic differentiation in Nouelia insignis (Asteraceae), a narrowly distributed and endemic species in China, revealed by ISSR fingerprinting

BACKGROUND AND AIMS: Nouelia insignis Franch., a monotypic genus of the Asteraceae, is an endangered species endemic in Yunnan and Sichuan Provinces of China. Most of the populations are seriously threatened. Some of them are even at the brink of extinction. In this study, the genetic diversity and differentiation between populations of this species were examined in two drainage areas. METHODS: DNA fingerprinting based on inter-simple sequence repeat polymorphisms was employed to detect the genetic variation and population structure in the species. KEY RESULTS: Genetic diversity at species level was high with P=65.05% (percentage of polymorphic loci) and Ht=0.2248 (total genetic diversity). The coefficient of genetic differentiation among populations, Gst, which was estimated by partitioning the total gene diversity, was 0.2529; whereas, the genetic differentiation between populations in the Jinsha and Nanpan drainage areas was unexpectedly low (Gst=0.0702). CONCLUSIONS: Based on the genetic analyses of the DNA fingerprinting, recent habitat fragmentation may not have led to genetic differentiation or the loss of genetic diversity in the rare species. Spatial apportionment of fingerprinting polymorphisms provides a footprint of historical migration across geographical barriers. The high diversity detected in this study holds promise for conservation and restoration efforts to save the endangered species from extinction.     

A keystone predator controls bacterial diversity in the pitcher-plant (Sarracenia purpurea) microecosystem

The community of organisms inhabiting the water-filled leaves of the carnivorous pitcher-plant Sarracenia purpurea includes arthropods, protozoa and bacteria, and serves as a model system for studies of food web dynamics. Despite the wealth of data collected by ecologists and zoologists on this food web, very little is known about the bacterial assemblage in this microecosystem. We used terminal restriction fragment length polymorphism (T-RFLP) analysis to quantify bacterial diversity within the pitchers as a function of pitcher size, pH of the pitcher fluid and the presence of the keystone predator in this food web, larvae of the pitcher-plant mosquito Wyeomyia smithii. Results were analysed at two spatial scales: within a single bog and across three isolated bogs. Pitchers were sterile before they opened and composition of the bacterial assemblage was more variable between different bogs than within bogs. Measures of bacterial richness and diversity were greater in the presence of W. smithii and increased with increasing pitcher size. Our results suggest that fundamental ecological concepts derived from macroscopic food webs can also be used to predict the bacterial assemblages in pitcher plants.     

Assessment of genetic diversity in Lepidium sativum (Chandrasur) a medicinal herb used in folklore remedies in India using RAPD

Lepidium sativum L. is a fast growing edible herb which belongs to family Brassicaceae. The seeds of L. sativum are aperient, diuretic, tonic, demulcent, carminative, galatogogue and emmenagogue. They have been used in the treatment of bacterial and fungal infections, as an aperient and also possess antibacterial and antifungal properties. The seeds of this plant possess rapid bone fracture healing ability. Despite of its diverse medicinal properties no molecular data for diversity analysis is available till date. During this study random amplified polymorphic DNA (RAPD) markers were used to detect genetic variations of L. sativum. Initially 50 decamer primers were screened, out of which only 32 primers showed reproducible fragments with easily recordable bands. A total of 414 reproducible and clear bands were distinguished across the selected primers and statistical analysis showed 361 polymorphic bands and 53 monomorphic bands. Cluster analysis of the genotypes based on UPGMA divided the 18 genotypes into two main clusters, with first cluster having only HCS-20 genotype of L. sativum and other having rest of all 17 genotypes. The dendrogram based on similarity matrix revealed 23–66% genetic relatedness among 18 genotypes. The results of the present study can be used for molecular breeding and improvement of L. sativum for various desired traits through hybridization in future.     

Genetic variability and founder effect in the pitcher plant Sarracenia purpurea (Sarraceniaceae) in populations introduced into Switzerland: from inbreeding to invasion

BACKGROUND AND AIMS: The long-lived and mainly outcrossing species Sarracenia purpurea has been introduced into Switzerland and become invasive. This creates the opportunity to study reactions to founder effect and how a species can circumvent deleterious effects of bottlenecks such as reduced genetic diversity, inbreeding and extinction through mutational meltdown, to emerge as a highly invasive plant. METHODS: A population genetic survey by random amplified polymorphism DNA markers (RAPD) together with historical insights and a field pollination experiment were carried out. KEY RESULTS: At the regional scale, S. purpurea shows low structure (thetast=0.072) due to a recent founder event and important subsequent growth. Nevertheless, multivariate statistical analyses reveal that, because of a bottleneck that shifted allele frequencies, most of the variability is independent among populations. In one population (Tenasses) the species has become invasive and genetic analysis reveals restricted gene flow and family structure (thetast=0.287). Although inbreeding appears to be high (Fis >0.410 from a Bayesian estimation), a field pollination experiment failed to detect significant inbreeding depression upon F1 seed number and seed weight fitness-traits. Furthermore, crosses between unrelated individuals produced F1 seeds with significantly reduced fitness, thus showing local outbreeding depression. CONCLUSIONS: The results suggest that, under restricted gene flow among families, the species may not only have rapidly purged deleterious alleles, but also have undergone some form of selection for inbreeding due to co-adaptation between loci.     

High genetic diversity in Taihangia rupestris Yu et Li,a rare cliff herb endemic to China,based on inter-simple sequence repeat markers

Taihangia rupestris Yu et Li is an endangered perennial herb endemic to China. Besides being at an important systematic position in the tribe Dryadeae, it also is a valuable wild flower resource. Inter-simple sequence repeat (ISSR) markers were employed to estimate the genetic diversity and genetic differentiation of ten populations. Relatively high genetic diversity was detected at species levels (PPB = 80.43; h = 0.2479; I = 0.3785). Most of the variation (73.51%) occurred within populations, and 26.49% was found among populations; Nei’s differentiation coefficient (Gst = 0.2435) supported this result. The Mantel test revealed a significantly positive correlation between geographical distance and genetic distance (r = 0.611, P < 0.001). The levels and patterns of genetic diversity of T. rupestris were assumed to result from its ancient origin and mixed reproductive modes. Based on these results, both ex situ and in situ conservation are proposed.     

High ecomorphological diversity among Early Cretaceous frogs from a large subtropical wetland of Iberia

Anurans are extremely diverse amphibians with a unique Bauplan, whose origin, early ecomorphological diversification, and adaptive significance remain elusive. Their early fossil record is improving at an accelerated pace worldwide, but its contribution to these issues is still wanting. Here we explore ecomorphological diversity among Early Cretaceous (Barremian) frogs that inhabited a large subtropical wetland from Iberia by inferring locomotor abilities with a phylogenetic flexible discriminant analysis on data from limb proportions, which strongly correlate with locomotion. The results show a remarkable diversity among these frogs when compared with the extant diversity from this region and from tropical and subtropical wetland assemblages worldwide, encompassing miniature to medium-size jumpers and dedicated swimmers with more extreme proportions than extant ‘archeobatrachians,’ but also more generalized jumping and/or swimming forms. This agrees with the inferred wetland paleoenvironment and the hypothesis regarding early frogs as small poor jumpers or swimmers that evolved their peculiar Bauplan in aquatic environments.     

Population structure and genetic diversity of Dysosma versipellis (Berberidaceae), a rare endemic from China

Dysosma versipellis (Berberidaceae) is an endangered and endemic species in China. To provide scientific foundation for formulating conservation strategies, we sampled six extant populations of this species and assessed the levels and patterns of genetic diversity using ISSR markers (11 primers). Of 144 bands detected 57.64% were polymorphic, but on average only 20.72% were polymorphic within populations. Our results revealed a low level of intraspecific genetic diversity (at population level: H_pop = 0.082, H_B = 0.177, SI = 0.1194; at species level: H_pop = 0.207, H_B = 0.378, SI = 0.3069). A high level of genetic differentiations among populations was detected based on Nei's genetic diversity analysis (60%), AMOVA analysis (65%), and Bayesian analysis (53%). The low levels of heterozygosity and high genetic differentiation observed in D. versipellis may be the consequence of low rate of natural recruitment, clonal growth, gene drift, and habitat fragmentation. Based on this, we suggest that in situ conservation be an important and practical measure for maintaining the genetic diversity of this species. Ex situ conservation should sample from different populations across the distribution range of the species to conserve high genetic diversity.