Community analysis of arbuscular mycorrhizal fungi associated with Ammophila arenaria in Dutch coastal sand dunes

A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) approach for the detection and characterization of arbuscular mycorrhizal fungi (AMF) 18S ribosomal DNA (rDNA) was developed and applied to the study of AMF communities associated with the main sand-stabilizing plant species of the Dutch sand dunes, marram grass (Ammophila arenaria, L.). DNA was extracted directly from plant roots, soil or isolated AMF spores, and prominent bands resulting from AMF-specific DGGE profiles were excised for sequence analysis. This strategy provided a robust means of detecting and identifying AMF-like species without the use of trap plant cultivation methods. A number of Glomus-like and Scutellospora-like sequences was detected, including a putatively novel Glomus species, and differences were observed in the dominant AMF-like populations detected in healthy vs. degenerating stands of A. arenaria and in bulk sand dune soil. It has previously been suggested that plant pathogens, such as fungi and nematodes, may contribute to the decline of A. arenaria. Although no causal relationship can be drawn between the observed differences in the dominantly detected AMF-like populations and the vitality of plant growth, these results indicate that mutualistic interactions between this plant and AMF should not be overlooked when examining the role of soil-borne microorganisms in vegetation dynamics. In addition, there were discrepancies observed between the AMF-like groups detected in spore populations vs. direct 18S rDNA analysis of root material, corroborating previous suggestions that spore inspection alone may poorly represent actual AMF population structure.     

Nitrogen fixation by Bacillus strains isolated from the rhizosphere of Ammophila arenaria

Summary Several Bacillus strains, from the rhizosphere of Ammophila arenaria, appeared on ‘nitrogen-free’ agar plates. They were able to grow in nitrogen-poor medium to which 0.1% yeast extract was added. Three of these bacilli were tested for their ability to fix nitrogen using the acetylene reduction assay. The C₂H₂-reducing activity was determined at 8-hour intervals during their growth cycle. C₂H₂ reduction (and accordingly N₂ fixation) was greater under anaerobic than aerobic conditions. Additions of 0.1% CaCO₃ significantly increased the C₂H₂-reducing activity under both conditions. Characterisation suggests that these strains are new nitrogen-fixing Bacillus species. re]19740121     

Adaptation in perennial coastal plants-with particular reference to heritable variation in Puccinellia maritima and Ammophila arenaria

Perennial species invading the early stages of primary successions face constant, and often rapid, change in their biotic and abiotic environment. The relative abilities of different species to adapt to this change is reflected in the zonation patterns which characterize coastal vegetation. Variation in those species with wide ecological amplitudes, particularly in populations near the boundary of the realized niche, is likely to be particularly revealing.The pattern of heritable variation in Puccinellia maritima on salt marshes indicates directional selection for traits increasing plant vigour and competitive ability; presumably the effect of increasing plant density. Adaptation is by both genetic differentiation and phenotypic flexibility, the former being evident in adjacent grazed and ungrazed marshes and the latter in a mosaic of tall and short vegetation types. By contrast variation in Ammophila arenaria on dunes exhibits high levels of phenotypic flexibility, growth in a range of environments indicating that plants from fore-dune populations are higher responders than those from mature dunes.Among the implications of these results, and by comparison with other species, is the fact that, ironically, niche expansion for some salt marsh perennials may require the evolution of an annual strategy, and that a Darwinian selection model may help to explain variation in Ammophila's apparent vigour in dunes of different age.Nomenclature follows Tutin et al.: Flora Europaea (1964–1980).I am grateful to several colleagues and students, past and present, who have helped with the studies of Puccinellia and Ammophila and I am particularly indebted in this respect to Richard Scott.     

Reduced competitive ability in an invasive plant

One explanation for successful plant invaders is that they evolved to be more competitive. An intuitive prediction of this Evolution of Increased Competitive Ability (EICA) hypothesis never previously tested is that invasive populations should outcompete their native ‘ancestors’ in a common environment. We tested this idea in a diallel competition experiment with Alliaria petiolata where offspring from native and invasive populations were grown alone or in all pairwise combinations. While without competition, there were no differences between native and invasive populations, native populations outperformed invasive ones when competing against each other. Our results contradict the EICA hypothesis and we conclude that it does not not hold for Alliaria petiolata. Instead, we formulate a new ERCA (Evolutionary Reduced Competitive Ability) hypothesis: if there is less competition in the invasive range and competitive ability involves traits that have a fitness cost, then selection might act against it, thereby reducing intraspecific interactions too.     

Biotic soil factors affecting the growth and development of Ammophila arenaria

Summary To study the origin of replant disease of Ammophila arenaria (L.) Link the growth and development in sand originating from the rhizosphere of a natural Ammophila vegetation was compared with the growth in sand from the sea-floor. In a greenhouse experiment, the growth of Ammophila seedlings in rhizosphere sand, when compared with that in sea sand, was significantly reduced. As sterilization by means of gamma-irradiation increased the biomass production of Ammophila seedlings significantly, it was concluded that the rhizosphere sand contained biotic factors that were harmful to Ammophila. In rhizosphere sand the roots of Ammophila were brown and poorly developed, and the specific uptake rates of N, P and K were reduced. The shoot weight proportion of the total plant dry matter was hardly influenced. In an outdoor experiment with Ammophila seedlings and cuttings, using both sands, the mortality was high and the plants were feeble in rhizosphere sand whereas plants in sea sand grew vigorously. It seems plausible that the plants in rhizophere sand were dessicated because the root system was shallow and badly developed. In the greenhouse experiments, Ammophila cuttings were less sensitive to the inhibiting factors in the rhizosphere than seedlings. This was confirmed in the outdoor experiment. Calammophila baltica (Fluegge ex Schrader) Brand, however, was hardly affected by the harmful biotic factors in the greenhouse. These results are discussed with reference to the ecology of Ammophila. It is assumed that the catching of fresh windblown sand provides Ammophila with a way to escape from harmful biotic soil factors, and it was concluded that degeneration of Ammophila is caused mainly by self-intolerance due to these biotic soil factors.     

Nutrient and productivity relations of the dune grasses Ammophila arenaria and Elymus mollis

Summary A comparative study of blade photosynthesis and nitrogen use efficiency was made on the dune grasses Ammophila arenaria and Elymus mollis. In the laboratory, an open system gas analysis apparatus was used to examine the gas exchange characteristics of blades as influenced by nitrogen supply. Plants were grown under near-ambient coastal conditions in a greenhouse near Bodega Bay, California, and given either high or low supplies of nitrogen in an otherwise complete nutrient solution. In the field, ¹⁴CO₂ uptake techniques were employed to measure the seasonal patterns of blade photosynthesis of plants growing in situ at Point Reyes National Seashore. Blades used in the lab and field studies were analyzed for total nitrogen content, thus allowing for calculations of photosynthetic nitrogen use efficiency (CO₂ fixed/unit of blade N.).Under laboratory conditions, the introduced Ammophila developed higher rates of light-saturated photosynthesis than the native Elymus, especially under the nitrogenlimited growth regime. Higher rates of photosynthesis and lower concentrations of blade N resulted in a significantly greater nitrogen use efficiency for Ammophila regardless of nutrient treatment. Low N availability induced qualitatively similar physiological responses in both species, including reductions in maximum net photosynthesis, mesophyll conductance, leaf conductance, dark respiration, and blade nitrogen content, and an increase in the CO₂ compensation point.Although the photosynthetic rates of Ammophila blades were higher in the lab, those of Elymus blades were consistently higher in the field. This could have resulted from differential effects of drought on the two species (i.e. Ammophila may have been more sensitive) or a higher photosynthetic capacity in Elymus that reflected the greater (1.2–1.5 X) nitrogen content of its blades. However, the nitrogen use efficiency of Ammophila blades was greater than that of Elymus throughout most of the sampling year, despite lower average rates of field photosynthesis.The results indicated that rates of photosynthesis perunit of blade area do not account for the greater aboveground productivity of Ammophila stands along the Pacific coast of North America. Instead, efficient nitrogen use in photosynthesis maycomplement other structural and physiological traits and thereby enhance long-term carbon gain in Ammophila relative to Elymus.     

Mechanism of control of root-feeding nematodes by mycorrhizal fungi in the dune grass Ammophila arenaria

Root-feeding herbivores can affect plant performance and the composition of natural plant communities, but there is little information about the mechanisms that control root herbivores in natural systems. This study explores the interactions between the pioneer dune grass Ammophila arenaria, arbuscular mycorrhizal fungi (AMF) and the root-feeding nematode Pratylenchus penetrans. Our objectives were to determine whether AMF can suppress nematode infection and reproduction and to explore the mechanisms of nematode control by AMF. A sequential inoculation experiment and a split-root experiment were designed to analyse the importance of plant tolerance and resistance and of direct competition between AMF and P. penetrans for the root herbivore and the plant. Root infection and multiplication of P. penetrans were significantly reduced by the native inoculum of AMF. Plant preinoculation with AMF further decreased nematode colonization and reproduction. Nematode suppression by AMF did not occur through a systemic plant response but through local mechanisms. Our results suggest that AMF are crucial for the control of root-feeding nematodes in natural systems and illustrate that locally operating mechanisms are involved in this process.     

Diversity of AMF associated with Ammophila arenaria ssp. arundinacea in Portuguese sand dunes

Dune vegetation is essential for the formation and preservation of sand dunes and the protection of the coast line. Coastal sand dunes are harsh environments where arbuscular mycorrhizal fungi (AMF) play an important role in promoting plant establishment and growth. We present a study of the diversity of AMF associated with A. arenaria ssp. arundinacea in two locations of the Portuguese coast under a Mediterranean climate. These two locations were selected to compare a well-preserved dune system from a protected area with a degraded dune system from a public beach. AMF diversity was assessed mainly by cloning and sequencing of a fragment of the ribosomal SSU using the primer NS31 and AM1. Most of the 89 AMF clones obtained from the rhizosphere and roots of A. arenaria belonged to the genus Glomus, the largest clade within the Glomeromycota. Higher AMF diversity was found in the least disturbed site, in which spores of Scutellospora persica, Glomus constrictum and Glomus globiferum were found in the rhizosphere of A. arenaria.     

Shift in competitive ability mediated by soil biota in an invasive plant

Understanding the shifts in competitive ability and its driving forces is key to predict the future of plant invasion. Changes in the competition environment and soil biota are two selective forces that impose remarkable influences on competitive ability. By far, evidence of the interactive effects of competition environment and soil biota on competitive ability of invasive species is rare. Here, we investigated their interactive effects using an invasive perennial vine, Mikania micrantha. The competitive performance of seven M. micrantha populations varying in their conspecific and heterospecific abundance were monitored in a greenhouse experiment, by manipulating soil biota (live and sterilized) and competition conditions (competition‐free, intraspecific, and interspecific competition). Our results showed that with increasing conspecific abundance and decreasing heterospecific abundance, (1) M. micrantha increased intraspecific competition tolerance and intra‐ vs. interspecific competitive ability but decreased interspecific competition tolerance; (2) M. micrantha increased tolerance of the negative soil biota effect; and (3) interspecific competition tolerance of M. micrantha was increasingly suppressed by the presence of soil biota, but intraspecific competition tolerance was less affected. These results highlight the importance of the soil biota effect on the evolution of competitive ability during the invasion process. To better control M. micrantha invasion, our results imply that introduction of competition‐tolerant native plants that align with conservation priorities may be effective where M. micrantha populations are long‐established and inferior in inter‐ vs. intraspecific competitive ability, whereas eradication may be effective where populations are newly invaded and fast‐growing.     

History of exposure to herbivores increases the compensatory ability of an invasive plant

Release from natural enemies is frequently cited as an important factor contributing to plant invasions. But such effects are likely to be temporary—native herbivores can form new plant-herbivore associations and co-evolved insects might reach the new range. While the potential effects of the initial enemy release have been well studied, the consequences of any resumption of herbivory are poorly understood. Alternanthera philoxeroides is one of the most widespread invasive plants in China and is attacked both by a specialist herbivore introduced from the native range, Agasicles hygrophila, and a native beetle Cassida piperata Hope which has formed a new association. However, these insects are not found throughout the invaded range. To test the effect of the history of population exposure to herbivory on compensatory ability, plants were cultured from 14 populations around China that differed in whether A. hygrophila or C. piperata were present. Treatment plants were exposed to herbivory by A. hygrophila for a week until 50% of the leaf area was defoliated, then grown for 80 days. Plants from populations with prior exposure to herbivory (of any kind) accumulated more root mass than populations without prior exposure, indicating that prior exposure to insects can stimulate plant compensation to herbivory. We would recommend that potential changes in plant tolerance in response to prior exposure to herbivory are considered in invasive plant management plans that employ bio-control agents.     

Woody plant colonization in an experimentally fragmented landscape

The pattern of distribution and abundance of woody plants colonizing old fields is influenced by landscape spatial features, in particular, by the distance from the old field to propagule sources and the size of the habitat patches undergoing succession. Colonization is also influenced by species life history traits, such as dispersal mode, growth form, and fecundity. As part of a long-term project studying effects of habitat fragmentation on secondary succession at the prairie-forest ecotone, we have examined the colonization patterns of early-successional woody plants in an experimentally fragmented old field, with emphasis on the three woody species [Cornus drummondii C. A. Mey (rough-leaved dogwood), Ulmus rubra Muhl. (slippery elm), and Juniperus virginiana L. (red cedar)], which currently dominate the woody community on the site. The shapes of the colonization curve (proportion of colonized quadrats vs time) differed between C. drummondii and U. rubra. The rate of colonization by C. drummondii showed a pattern of acceleration after its initial colonization, consistent with rapid in situ recruitment from clonal growth and early seed production. By contrast, colonization by U. rubra fits a roughly linear pattern, consistent with recruitment only from external propagule sources. For both C. drummondii and U. rubra, density is currently greater in large patches than in small patches. No patch size difference was found for J. virginiana. The stern density of both C. drummondii and U. rubra exponentially decreased with distance to external propagule sources. The negative exponential pattern of U. rubra (wind-dispersed) with distance is sharper than that of C drummondii (bird-dispersed). Moreover, the amount of spatial variation in density explained by distance to source is greater on small patches. Our results highlight the importance of life history traits of colonizing species and spatial aspects of habitat during succession.     

Increased compensatory ability of an invasive plant to above- and below-ground enemies in monocultures

Above- and below-ground enemies have prominent influence on plant invasions, and increasing evidence has shown that plant invasions are also affected by inter- or intraspecific interactions between individual plants. However, how these two factors interactively affect plant invasions has rarely been tested. Here, we examined the response of the invasive plant Alternanthera philoxeroides (Mart.) Griseb. to above- and below-ground enemies at varying plant densities in a greenhouse experiment in Wuhan, China. We found that both above- and below-ground enemies decreased the plant total and root mass at individual and population levels, but that of the two, below-ground enemies had a greater effect than above-ground enemies, and that the two guilds of enemies acted additively on the plant. However, their impacts decreased as the plant density increased, due to enhanced plant tolerance to both guilds of enemies. The increased plant tolerance may result from changes in plant resource allocation patterns, corresponding to a positive linear relationship between the ratio of fine root mass to total root mass and plant density. Given that forming dense monocultures in their new ranges is one of the most important characteristics of invasive plants, we propose that the high compensatory ability at dense monocultures may be an important mechanism underlying exotic species invasion.     

Allozyme diversity and geographic variation in the widespread coastal sedge, Carex arenaria

Abstract. Allozyme electrophoresis was used to investigate the structure of genetic variation in the rhizomatous coastal sedge, Carex arenaria, throughout its European range — from the SW Iberian peninsula to the Baltic region. Material was sampled from 77 sites in five geographic regions. Nine of the 13 investigated loci were polymorphic in the total material and there were interregional differences in the number of polymorphic loci per site and the percentage of variable sites. In the Scandinavia/Baltic region only 61% of the sites contained at least one locus with more than one allele, whereas all the British and SW Iberian sites were variable. There was a general tendency for the regional frequencies of the less common alleles at individual loci to decline from SW to NE. The mean (over loci and sites) within‐site gene diversity (H _site) was 0.064 (in calculations based on the number of observed multilocus allozyme genotypes within each sampling site). Although there was considerable variation between geographically adjacent sites, within‐site diversity showed a general decrease from SW to NE in Europe. There were significant differences in within‐region gene diversity (H_reg) for the four most variable loci between the five regions. H_reg generally decreased from SW to NE Europe and most loci showed the highest diversity in the SW Iberian peninsula and the Bay of Biscay regions. The mean (over loci) gene diversity in the total material (H_tot) was 0.070 and the levels of diversity in Carex arenaria are substantially lower than is usual in rhizomatous sedges. The within‐site, between‐site and between‐regional components of the total diversity were 92.4%, 2.5% and 5.1%, respectively. The low levels of overall gene diversity in C. arenaria and the successive decrease in diversity from SW to NE are interpreted in terms of the species’ history of postglacial spread into northern Europe. Despite the overall northwards decrease in diversity, the widespread occurrence of less common alleles and the lack of regional deviations from Hardy–Weinberg genotype frequency expectations suggest that C. arenaria is not predominantly self‐fertilized.     

Resource utilization conditions as biochar of an invasive plant Spartina alterniflora in coastal wetlands of China

Converting feedstocks of invasive plants into biochar is a new and cost‐effective measure for their control, and benefits for the sustainable development of native ecosystems. Spartina alterniflora, an invasive plant widely distributed in coastal wetlands of China, was used to produce biochar. We aimed to analyze how S. alterniflora biochar properties changed with desalination of feedstocks, pyrolysis temperature, and residence time. Results showed that desalting feedstocks increased biochar pH, stability, porosity, and surface area, but diminished biochar yield and polarity. Pyrolysis temperature positively affected biochar pH, surface area, and pore volume, while it had negative effects on biochar yield, oxygen and hydrogen contents, hydrogen/carbon and oxygen/carbon ratios, pore size, and function groups. However, residence time of pyrolysis had slight effects on biochar properties. The results are valuable for optimizing pyrolysis temperature and pretreatment measure of feedstocks, to tune S. alterniflora biochar properties for specific environmental usage.     

Interactions between root and shoot herbivores of Ammophila arenaria in the laboratory do not translate into correlated abundances in the field

Over the past decades a growing body of literature has presented proof of the possible interactions between foliar and root herbivores. These effects can be positive, negative or neutral in either direction, depending on the species and the involved mechanism. Most of these studies however concern experiments under controlled conditions. Whether these interactions affect the distribution of herbivores under natural conditions still largely remains an open question. This study examined interactions between root feeding nematodes and shoot feeding aphids on Ammophila arenaria in the laboratory. We subsequently addressed the question whether expectations from this experiment are reflected in correlations between plant related variables and the abundance of both herbivores in the field. We demonstrated that nematodes and aphids can negatively affect each other in a controlled microcosm. In the field however no significant correlations between nematode and aphid abundances could be detected. There, shorter plants with a more vital leaf set and a higher root density supported the highest numbers of aphids. Plants with a lower root density and higher root vitality held more migratory endoparasitic nematodes, while more nematode cysts were found among roots with a low vitality. A certain plant property can furthermore affect above‐ and belowground herbivores in the opposite direction, such as root density in this case. This study suggests that effects of root herbivores on foliar herbivores or vice versa seem to be blurred in a field situation where other variables related to plant vitality and water content structure the herbivore populations. Therefore, caution should be used in generalising the prevalence of these interactions between the above‐ and belowground fauna, based solely on laboratory experiments.     

Comparison of the herbivore defense and competitive ability of ancestral and modern genotypes of an invasive plant,Lespedeza cuneata

The evolution of increased competitive ability (EICA) hypothesis provides a compelling explanation for the success of invasive species. It contends that because alien plants have escaped their coevolved natural enemies, selection pressures favor a diversion of resources from herbivore defense to traits that confer increased competitive ability. Here, we provide evidence for EICA in the noxious grassland invader Lespedeza cuneata, by comparing the ancestral genotype introduced to North America in 1930 with modern‐day invasive (North American) and native (Japanese) genotypes. We found that the invasive genotype was a better competitor than either the native or the ancestral genotype. Further, the invasive genotype exhibited greater induced resistance but lower constitutive resistance than the ancestral and native genotypes. Our results suggest that selection has played a pivotal role in shaping this invasive plant species into a more aggressive, but less constitutively defended competitor.