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1.  Silicon’s organic pool and biological cycle in moso bamboo community of Wuyishan Biosphere Reserve*  
Biomineralization of Si by plants into phytolith formation and precipitation of Si into clays during weathering are two important processes of silicon’s biogeochemical cycle. As a silicon-accumulating plant, the widely distributed and woody Phyllostachys heterocycla var. pubescens (moso bamboo) contributes to storing silicon by biomineralization and, thus, prevents eutrophication of nearby waterbodies through silicon’s erosion of soil particles.
A study on the organic pool and biological cycle of silicon (Si) of the moso bamboo community was conducted in Wuyishan Biosphere Reserve, China. The results showed that: (1) the standing crop of the moso bamboo community was 13355.4 g/m2, of which 53.61%, 45.82% and 0.56% are represented by the aboveground and belowground parts of moso bamboos, and the understory plants, respectively; (2) the annual net primary production of the community was 2887.1 g/(m2·a), among which the aboveground part, belowground part, litterfalls, and other fractions, accounted for 55.86%, 35.30%, 4.50% and 4.34%, respectively; (3) silicon concentration in stem, branch, leaf, base of stem, root, whip of bamboos, and other plants was 0.15%, 0.79%, 3.10%, 4.40%, 7.32%, 1.52% and 1.01%, respectively; (4) the total Si accumulated in the standing crop of moso bamboo community was 448.91 g/m2, with 99.83% of Si of the total community stored in moso bamboo populations; (5) within moso bamboo community, the annual uptake, retention, and return of Si were 95.75, 68.43, 27.32 g/(m2·a), respectively; (6) the turnover time of Si, which is the time an average atom of Si remains in the soil before it is recycled into the trees or shrubs, was 16.4 years; (7) the enrichment ratio of Si in the moso bamboo community, which is the ratio of the mean concentration of nutrients in the net primary production to the mean concentration of nutrients in the biomass of a community, was 0.64; and lastly, (8) moso bamboo plants stored about 1.26×1010 kg of silicon in the organic pool made up by the moso bamboo forests in the subtropical area of China.
doi:10.1631/jzus.2006.B0849
PMCID: PMC1635818  PMID: 17048297
Phyllostachys heterocycla var. pubescens; Moso bamboo community; Silicon-accumulating; Silicon biological cycle; Wuyishan Biosphere Reserve
2.  The Root Herbivore History of the Soil Affects the Productivity of a Grassland Plant Community and Determines Plant Response to New Root Herbivore Attack 
PLoS ONE  2013;8(2):e56524.
Insect root herbivores can alter plant community structure by affecting the competitive ability of single plants. However, their effects can be modified by the soil environment. Root herbivory itself may induce changes in the soil biota community, and it has recently been shown that these changes can affect plant growth in a subsequent season or plant generation. However, so far it is not known whether these root herbivore history effects (i) are detectable at the plant community level and/or (ii) also determine plant species and plant community responses to new root herbivore attack. The present greenhouse study determined root herbivore history effects of click beetle larvae (Elateridae, Coleoptera, genus Agriotes) in a model grassland plant community consisting of six common species (Achillea millefolium, Plantago lanceolata, Taraxacum officinale, Holcus lanatus, Poa pratensis, Trifolium repens). Root herbivore history effects were generated in a first phase of the experiment by growing the plant community in soil with or without Agriotes larvae, and investigated in a second phase by growing it again in the soils that were either Agriotes trained or not. The root herbivore history of the soil affected plant community productivity (but not composition), with communities growing in root herbivore trained soil producing more biomass than those growing in untrained soil. Additionally, it influenced the response of certain plant species to new root herbivore attack. Effects may partly be explained by herbivore-induced shifts in the community of arbuscular mycorrhizal fungi. The root herbivore history of the soil proved to be a stronger driver of plant growth on the community level than an actual root herbivore attack which did not affect plant community parameters. History effects have to be taken into account when predicting the impact of root herbivores on grasslands.
doi:10.1371/journal.pone.0056524
PMCID: PMC3575479  PMID: 23441201
3.  Large herbivores may alter vegetation structure of semi-arid savannas through soil nutrient mediation 
Oecologia  2011;165(4):1095-1107.
In savannas, the tree–grass balance is governed by water, nutrients, fire and herbivory, and their interactions. We studied the hypothesis that herbivores indirectly affect vegetation structure by changing the availability of soil nutrients, which, in turn, alters the competition between trees and grasses. Nine abandoned livestock holding-pen areas (kraals), enriched by dung and urine, were contrasted with nearby control sites in a semi-arid savanna. About 40 years after abandonment, kraal sites still showed high soil concentrations of inorganic N, extractable P, K, Ca and Mg compared to controls. Kraals also had a high plant production potential and offered high quality forage. The intense grazing and high herbivore dung and urine deposition rates in kraals fit the accelerated nutrient cycling model described for fertile systems elsewhere. Data of a concurrent experiment also showed that bush-cleared patches resulted in an increase in impala dung deposition, probably because impala preferred open sites to avoid predation. Kraal sites had very low tree densities compared to control sites, thus the high impala dung deposition rates here may be in part driven by the open structure of kraal sites, which may explain the persistence of nutrients in kraals. Experiments indicated that tree seedlings were increasingly constrained when competing with grasses under fertile conditions, which might explain the low tree recruitment observed in kraals. In conclusion, large herbivores may indirectly keep existing nutrient hotspots such as abandoned kraals structurally open by maintaining a high local soil fertility, which, in turn, constrains woody recruitment in a negative feedback loop. The maintenance of nutrient hotspots such as abandoned kraals by herbivores contributes to the structural heterogeneity of nutrient-poor savanna vegetation.
doi:10.1007/s00442-010-1899-3
PMCID: PMC3057003  PMID: 21225433
Bush encroachment; Tree–grass competition; Nutrient hotspot; Seedling; Predation
4.  Sensitivity of the Invasive Geophyte Oxalis pes-caprae to Nutrient Availability and Competition 
Annals of Botany  2007;99(4):637-645.
Background and Aims
Invasion by alien plants may be partially related to disturbance-related increases in nutrient availability and decreases of competition with native species, and to superior competitive ability of the invader. Oxalis pes-caprae is an invasive winter geophyte in the Mediterranean Islands that reproduces vegetatively via bulbs. An investigation was made into the relative responses of O. pes-caprae and the native annual grass Lolium rigidum to nutrient availability and to competition with each other in order to understand patterns of invasion in the field. Because Oxalis accumulates oxalic acid in its leaves, which could ameliorate soil phosphorous availability, field observations were made to determine whether the presence of Oxalis alters soil P availability.
Methods
A full-factorial glasshouse experiment was conducted with nutrient availability (high and low) and competition (Lolium alone, Oxalis alone, and Lolium and Oxalis together). Plant performance was assessed by determining (1) above- and below-ground biomass at the time of Oxalis maximum biomass and (2) reproductive output of Oxalis and Lolium at the end of their respective growth cycles. Measurements were also taken for leaf N and P content. Soil samples were taken in the field from paired Oxalis-invaded and non-invaded plots located in Menorca (Balearic Islands) and available P was determined.
Key Results
High nutrient availability increased Oxalis and Lolium vegetative biomass and reproductive output to a similar degree. Competition with Lolium had a much stronger negative effect on Oxalis bulb production than reduced nutrients. Lolium was a superior competitor than Oxalis; the latter did not affect Lolium maximum biomass and spike production. Significantly greater soil-P availability in Oxalis-invaded field soils relative to paired non-invaded soils suggest that Oxalis influences soil P cycling.
Conclusions
Oxalis is a poor competitor. This is consistent with the preferential distribution of Oxalis in disturbed areas such as ruderal habitats, and might explain its low influence on the cover of native species in invaded sites. The results also suggest that certain disturbances (e.g. autumn ploughing) may greatly enhance Oxalis invasion.
doi:10.1093/aob/mcl289
PMCID: PMC2802929  PMID: 17298990
Oxalis pes-caprae; invasive species; competition; nutrients; Lolium rigidum; asexual reproduction; Mediterranean Islands
5.  Olivine Weathering in Soil, and Its Effects on Growth and Nutrient Uptake in Ryegrass (Lolium perenne L.): A Pot Experiment 
PLoS ONE  2012;7(8):e42098.
Mineral carbonation of basic silicate minerals regulates atmospheric CO2 on geological time scales by locking up carbon. Mining and spreading onto the earth's surface of fast-weathering silicates, such as olivine, has been proposed to speed up this natural CO2 sequestration (‘enhanced weathering’). While agriculture may offer an existing infrastructure, weathering rate and impacts on soil and plant are largely unknown. Our objectives were to assess weathering of olivine in soil, and its effects on plant growth and nutrient uptake. In a pot experiment with perennial ryegrass (Lolium perenne L.), weathering during 32 weeks was inferred from bioavailability of magnesium (Mg) in soil and plant. Olivine doses were equivalent to 1630 (OLIV1), 8150, 40700 and 204000 (OLIV4) kg ha−1. Alternatively, the soluble Mg salt kieserite was applied for reference. Olivine increased plant growth (+15.6%) and plant K concentration (+16.5%) in OLIV4. At all doses, olivine increased bioavailability of Mg and Ni in soil, as well as uptake of Mg, Si and Ni in plants. Olivine suppressed Ca uptake. Weathering estimated from a Mg balance was equivalent to 240 kg ha−1 (14.8% of dose, OLIV1) to 2240 kg ha−1 (1.1%, OLIV4). This corresponds to gross CO2 sequestration of 290 to 2690 kg ha−1 (29 103 to 269 103 kg km−2.) Alternatively, weathering estimated from similarity with kieserite treatments ranged from 13% to 58% for OLIV1. The Olsen model for olivine carbonation predicted 4.0% to 9.0% weathering for our case, independent of olivine dose. Our % values observed at high doses were smaller than this, suggesting negative feedbacks in soil. Yet, weathering appears fast enough to support the ‘enhanced weathering’ concept. In agriculture, olivine doses must remain within limits to avoid imbalances in plant nutrition, notably at low Ca availability; and to avoid Ni accumulation in soil and crop.
doi:10.1371/journal.pone.0042098
PMCID: PMC3415406  PMID: 22912685
6.  The Terrestrial Silica Pump 
PLoS ONE  2012;7(12):e52932.
Silicon (Si) cycling controls atmospheric CO2 concentrations and thus, the global climate, through three well-recognized means: chemical weathering of mineral silicates, occlusion of carbon (C) to soil phytoliths, and the oceanic biological Si pump. In the latter, oceanic diatoms directly sequester 25.8 Gton C yr−1, accounting for 43% of the total oceanic net primary production (NPP). However, another important link between C and Si cycling remains largely ignored, specifically the role of Si in terrestrial NPP. Here we show that 55% of terrestrial NPP (33 Gton C yr−1) is due to active Si-accumulating vegetation, on par with the amount of C sequestered annually via marine diatoms. Our results suggest that similar to oceanic diatoms, the biological Si cycle of land plants also controls atmospheric CO2 levels. In addition, we provide the first estimates of Si fixed in terrestrial vegetation by major global biome type, highlighting the ecosystems of most dynamic Si fixation. Projected global land use change will convert forests to agricultural lands, increasing the fixation of Si by land plants, and the magnitude of the terrestrial Si pump.
doi:10.1371/journal.pone.0052932
PMCID: PMC3534122  PMID: 23300825
7.  Impacts of seedling herbivory on plant competition and implications for species coexistence 
Annals of Botany  2009;103(8):1347-1353.
Background and Aims
Although the causes and consequences of seedling herbivory for plant community composition are well understood, the mechanisms by which herbivores influence plant species recruitment to the established phase remain less clear. The aim was to examine how variation in the intensity of seedling herbivory interacts with growth-defence trade-offs and herbivore feeding preferences to affect plant community development.
Methods
Using 14-d-old seedlings of Trifolium pratense and T. repens, relative growth and susceptibility to herbivory by the snail Helix aspersa was quantified to elucidate putative growth-defence trade-offs for these species. Then mixed assemblages of 14-d-old Trifolium seedlings were exposed to herbivory by zero, two, five or ten snails and determined how variation in the intensity of herbivory affected competitive interactions into the mature phase (as measured by total plant biomass at 120 d old).
Key Results
In the absence of herbivory, communities were dominated by T. pratense; a result expected on the basis that it yielded larger and presumably more competitive seedlings. However, when seedlings were exposed to herbivory, the balance of competition shifted. At low levels of herbivory (two snails), both Trifolium species contributed equally to total plant biomass. More intense herbivory (five snails) resulted in almost total mortality of T. pratense and dominance of the mature community by T. repens. The most intense herbivory (ten snails) effectively removed all seedlings from the experimental community.
Conclusions
The study illustrates a mechanism whereby spatio-temporal fluctuations in seedling herbivory, when coupled with species-specific variation in competitive ability and sensitivity to herbivore attack, can differentially influence plant recruitment into the mature phase. This mechanism may be a key element in our attempts to understand plant species coexistence, since fluctuations in plant recruitment are fundamental to the many theories that view coexistence as a consequence of a spatio-temporal lottery for dominance over regeneration micro-sites.
doi:10.1093/aob/mcp081
PMCID: PMC2685311  PMID: 19351683
Growth-defence trade-off; lottery models; plant–animal interactions; plant size variability; seedling acceptability; seedling defence; spatio-temporal niches; Trifolium pratense; Trifolium repens
8.  Animal or Plant: Which Is the Better Fog Water Collector? 
PLoS ONE  2012;7(4):e34603.
Occasional fog is a critical water source utilised by plants and animals in the Namib Desert. Fog basking beetles (Onymacris unguicularis, Tenebrionidae) and Namib dune bushman grass (Stipagrostris sabulicola, Poaceae) collect water directly from the fog. While the beetles position themselves optimally for fog water collection on dune ridges, the grass occurs predominantly at the dune base where less fog water is available. Differences in the fog-water collecting abilities in animals and plants have never been addressed. Here we place beetles and grass side-by-side in a fog chamber and measure the amount of water they collect over time. Based on the accumulated amount of water over a two hour period, grass is the better fog collector. However, in contrast to the episodic cascading water run-off from the grass, the beetles obtain water in a steady flow from their elytra. This steady trickle from the beetles' elytra to their mouth could ensure that even short periods of fog basking – while exposed to predators – will yield water. Up to now there is no indication of specialised surface properties on the grass leafs, but the steady run-off from the beetles could point to specific property adaptations of their elytra surface.
doi:10.1371/journal.pone.0034603
PMCID: PMC3318004  PMID: 22509331
9.  Soil Microorganisms Alleviate the Allelochemical Effects of a Thyme Monoterpene on the Performance of an Associated Grass Species 
PLoS ONE  2011;6(11):e26321.
Background
Plant allelochemicals released into the soil can significantly impact the performance of associated plant species thereby affecting their competitive ability. Soil microbes can potentially affect the interaction between plant and plant chemicals by degrading the allelochemicals. However, most often plant-plant chemical interactions are studied using filter paper bioassays examining the pair-wise interaction between a plant and a plant chemical, not taking into account the potential role of soil microorganisms.
Methodology/Principal findings
To explore if the allelopathic effects on a grass by the common thyme monoterpene “carvacrol” are affected by soil microorganisms. Seedlings of the grass Agrostis capillaris originating from 3 different thyme sites were raised in the greenhouse. Seedlings were grown under four different soil treatments in a 2*2 fully factorial experiment. The monoterpene carvacrol was either added to standard greenhouse soil or left out, and soil was either sterilized (no soil microorganisms) or not (soil microorganisms present in soil). The presence of carvacrol in the soil strongly increased mortality of Agrostis plants, and this increase was highest on sterile soil. Plant biomass was reduced on soil amended with carvacrol, but only when the soil was also sterilized. Plants originating from sites where thyme produces essential oils containing mostly carvacrol had higher survival on soil treated with that monoterpene than plants originating from a site where thyme produced different types of terpenes, suggesting an adaptive response to the locally occurring terpene.
Conclusions/Significance
The study shows that presence of soil microorganisms can alleviate the negative effect of a common thyme monoterpene on the performance of an associated plant species, emphasizing the role of soil microbes in modulating plant-plant chemical interactions.
doi:10.1371/journal.pone.0026321
PMCID: PMC3219634  PMID: 22125596
10.  Rapid Plant Identification Using Species- and Group-Specific Primers Targeting Chloroplast DNA 
PLoS ONE  2012;7(1):e29473.
Plant identification is challenging when no morphologically assignable parts are available. There is a lack of broadly applicable methods for identifying plants in this situation, for example when roots grow in mixture and for decayed or semi-digested plant material. These difficulties have also impeded the progress made in ecological disciplines such as soil- and trophic ecology. Here, a PCR-based approach is presented which allows identifying a variety of plant taxa commonly occurring in Central European agricultural land. Based on the trnT-F cpDNA region, PCR assays were developed to identify two plant families (Poaceae and Apiaceae), the genera Trifolium and Plantago, and nine plant species: Achillea millefolium, Fagopyrum esculentum, Lolium perenne, Lupinus angustifolius, Phaseolus coccineus, Sinapis alba, Taraxacum officinale, Triticum aestivum, and Zea mays. These assays allowed identification of plants based on size-specific amplicons ranging from 116 bp to 381 bp. Their specificity and sensitivity was consistently high, enabling the detection of small amounts of plant DNA, for example, in decaying plant material and in the intestine or faeces of herbivores. To increase the efficacy of identifying plant species from large number of samples, specific primers were combined in multiplex PCRs, allowing screening for multiple species within a single reaction. The molecular assays outlined here will be applicable manifold, such as for root- and leaf litter identification, botanical trace evidence, and the analysis of herbivory.
doi:10.1371/journal.pone.0029473
PMCID: PMC3257244  PMID: 22253728
11.  Studies on the Host Range, Biology, and Pathogenicity of Punctodera punctata Infecting Turfgrasses 
Journal of Nematology  1985;17(2):162-165.
Punctodera punctata completed its life cycle on Poa annua (annual bluegrass), P. pratensis (Merion Kentucky bluegrass), Lolium perenne (perennial ryegrass), and Festuca rubra rubra (spreading fescue). Minimum time for completion of a life cycle from second-stage juvenile to mature brown cyst was 40 days at 22-28 C. Inoculation by single juveniles indicated that reproduction was most likely by amphimixis. Infestation levels of 50 or 500 juveniles/250 cm³ soil did not affect top dry weight, root dry weight, or total dry weight of Poa annua.
PMCID: PMC2618432  PMID: 19294077
cyst nematode; Poa annua; P. pratensis; Lolium perenne; Festuca rubra rubra; pathogenicity; life cycle; reproduction
12.  Grasshopper oral secretions increase salicylic acid and abscisic acid levels in wounded leaves of Arabidopsis thaliana 
Plant Signaling & Behavior  2011;6(9):1256-1258.
Recent investigations showed that the model plant Arabidopsis thaliana specifically responds to herbivory-associated molecular patterns by activating a sophisticated signaling network. The lipase activity of insect oral secretions was shown to elevate oxylipin levels when applied to puncture wounds in leaves. The results also demonstrated that the oral secretions of the generalist Schistocerca gregaria contained other, probably non-proteinous, elicitors of plant defense responses which induced mitogen-activated protein kinases, calcium signaling and ethylene levels.1 This addendum presents data on the levels of additional phytohormones that are elevated after application of S. gregaria oral secretion to wounded leaves. Abscisic acid and salicylic acid levels are significantly elevated after elicitation with S. gregaria oral secretions, adding another layer of complexity to the herbivory-induced response of A. thaliana.
doi:10.4161/psb.6.9.16552
PMCID: PMC3258045  PMID: 21847018
abscisic acid; Arabidopsis; herbivory; salicylic acid; Schistocerca gregaria
13.  Phytolith Assemblages and Systematic Associations in Grassland Species of the South-Eastern Pampean Plains, Argentina 
Annals of Botany  2006;98(6):1155-1165.
• Background and Aims Phytolith descriptions of South American plant species are scarce. This knowledge is crucial for the interpretation of the fossil phytolith record of a region. In this study phytolith assemblages and systematic relationships of the main grasses and Asteraceae species of Paspalum quadrifarium grassland are described.
• Methods Phytoliths from leaves of Poaceae and Asteraceae species were extracted by using a calcination technique. For each species, 350–400 phytoliths were counted and the relative frequency of each morphotype was calculated. Phytolith assemblages were subject to principal components analysis (PCA) and cluster analysis (Morisita index).
• Key Results PCA and cluster analysis showed family (Poaceae vs. Asteraceae) and subfamily (within Poaceae) differentiation. Exceptions to general trends described for other species were detected. Floristic variants of Paspalum quadrifarium grassland can be separated by their phytolith assemblages.
• Conclusions The study provides a reference collection of phytolith assemblages of the main Poaceae and Asteraceae species of Paspalum quadrifarium grassland and describes some phytolith morphotypes/systematic relationships useful for the analysis of fossil phytolith assemblages of the Pampean region.
doi:10.1093/aob/mcl207
PMCID: PMC3292269  PMID: 17030553
Paspalum quadrifarium; Asteraceae; Poaceae; phytolith assemblages; Argentina; Southern Pampa
14.  The skin prick test – European standards 
Skin prick testing is an essential test procedure to confirm sensitization in IgE-mediated allergic disease in subjects with rhinoconjunctivitis, asthma, urticaria, anapylaxis, atopic eczema and food and drug allergy. This manuscript reviews the available evidence including Medline and Embase searches, abstracts of international allergy meetings and position papers from the world allergy literature. The recommended method of prick testing includes the appropriate use of specific allergen extracts, positive and negative controls, interpretation of the tests after 15 – 20 minutes of application, with a positive result defined as a wheal ≥3 mm diameter. A standard prick test panel for Europe for inhalants is proposed and includes hazel (Corylus avellana), alder (Alnus incana), birch (Betula alba), plane (Platanus vulgaris), cypress (Cupressus sempervirens), grass mix (Poa pratensis, Dactilis glomerata, Lolium perenne, Phleum pratense, Festuca pratensis, Helictotrichon pretense), Olive (Olea europaea), mugwort (Artemisia vulgaris), ragweed (Ambrosia artemisiifolia), Alternaria alternata (tenuis), Cladosporium herbarum, Aspergillus fumigatus, Parietaria, cat, dog, Dermatophagoides pteronyssinus, Dermatophagoides farinae, and cockroach (Blatella germanica). Standardization of the skin test procedures and standard panels for different geographic locations are encouraged worldwide to permit better comparisons for diagnostic, clinical and research purposes.
doi:10.1186/2045-7022-3-3
PMCID: PMC3565910  PMID: 23369181
Sensitization; Inhalant allergens; Skin prick test panel; Aallergies; Type I allergy; Diagnostic test; Asthma
15.  Mass spectrometric analysis of electrophoretically separated allergens and proteases in grass pollen diffusates 
Respiratory Research  2003;4(1):10.
Background
Pollens are important triggers for allergic asthma and seasonal rhinitis, and proteases released by major allergenic pollens can injure airway epithelial cells in vitro. Disruption of mucosal epithelial integrity by proteases released by inhaled pollens could promote allergic sensitisation.
Methods
Pollen diffusates from Kentucky blue grass (Poa pratensis), rye grass (Lolium perenne) and Bermuda grass (Cynodon dactylon) were assessed for peptidase activity using a fluorogenic substrate, as well as by gelatin zymography. Following one- or two-dimensional gel electrophoresis, Coomassie-stained individual bands/spots were excised, subjected to tryptic digestion and analysed by mass spectrometry, either MALDI reflectron TOF or microcapillary liquid chromatography MS-MS. Database searches were used to identify allergens and other plant proteins in pollen diffusates.
Results
All pollen diffusates tested exhibited peptidase activity. Gelatin zymography revealed high Mr proteolytic activity at ~ 95,000 in all diffusates and additional proteolytic bands in rye and Bermuda grass diffusates, which appeared to be serine proteases on the basis of inhibition studies. A proteolytic band at Mr ~ 35,000 in Bermuda grass diffusate, which corresponded to an intense band detected by Western blotting using a monoclonal antibody to the timothy grass (Phleum pratense) group 1 allergen Phl p 1, was identified by mass spectrometric analysis as the group 1 allergen Cyn d 1. Two-dimensional analysis similarly demonstrated proteolytic activity corresponding to protein spots identified as Cyn d 1.
Conclusion
One- and two-dimensional electrophoretic separation, combined with analysis by mass spectrometry, is useful for rapid determination of the identities of pollen proteins. A component of the proteolytic activity in Bermuda grass diffusate is likely to be related to the allergen Cyn d 1.
PMCID: PMC239031  PMID: 14577842
16.  Development of a Genomic Microsatellite Library in Perennial Ryegrass (Lolium perenne) and its Use in Trait Mapping 
Annals of Botany  2008;101(6):845-853.
Background and Aims
Perennial ryegrass (Lolium perenne) is one of the key forage and amenity grasses throughout the world. In the UK it accounts for 70 % of all agricultural land use with an estimated farm gate value of £6 billion per annum. However, in terms of the genetic resources available, L. perenne has lagged behind other major crops in Poaceae. The aim of this project was therefore the construction of a microsatellite-enriched genomic library for L. perenne to increase the number of genetic markers available for both marker-assisted selection in breeding programmes and gene isolation.
Methods
Primers for 229 non-redundant microsatellite markers were designed and used to screen two L. perenne genotypes, one amenity and one forage. Of the 229 microsatellites, 95 were found to show polymorphism between amenity and forage genotypes. A selection of microsatellite primers was selected from these 95 and used to screen two mapping populations derived from intercrossing and backcrossing the two forage and amenity grass genotypes.
Key Results and Conclusions
The utility of the resulting genetic maps for analysis of the genetic control of target traits was demonstrated by the mapping of genes associated with heading date to linkage groups 4 and 7.
doi:10.1093/aob/mcn016
PMCID: PMC2710216  PMID: 18281692
Microsatellites; Lolium perenne; perennial ryegrass; trait mapping
17.  Density-dependent aposematism in the desert locust. 
The ecological processes underlying locust swarm formation are poorly understood. Locust species exhibit phenotypic plasticity in numerous morphological, physiological and behavioural traits as their population density increases. These density-dependent changes are commonly assumed to be adaptations for migration under heterogeneous environmental conditions. Here we demonstrate that density-dependent nymphal colour change in the desert locust Schistocerca gregaria (Orthoptera: Acrididae) results in warning coloration (aposematism) when the population density increases and locusts consume native, toxic host plants. Fringe-toed lizards (Acanthodactylus dumerili (Lacertidae)) developed aversions to high-density-reared (gregarious-phase) locusts fed Hyoscyamus muticus (Solanaceae). Lizards associated both olfactory and visual cues with locust unpalatability, but only gregarious-phase coloration was an effective visual warning signal. The lizards did not associate low rearing density coloration (solitarious phase) with locust toxicity. Predator learning of density-dependent warning coloration results in a marked decrease in predation on locusts and may directly contribute to outbreaks of this notorious pest.
PMCID: PMC1690497  PMID: 10670954
18.  Transport of silicon from roots to panicles in plants 
Silicon (Si) is the most abundant minerals in soil and exerts beneficial effects on plant growth by alleviating various stresses. The transport of Si from soil to the panicles is mediated by different transporters. Lsi1, belonging to a NIP group of the aquaporin family, is responsible for the uptake of Si from soil into the root cells in both dicots and monocots although its expression patterns and cellular localization differ with plant species. The subsequent transport of Si out of the root cells towards the stele is medicated by an active efflux transporter, Lsi2. Lsi1 and Lsi2 are polarly localized at the distal and proximal sides, respectively, of both exodermis and endodermis in rice root. Silicon in the xylem sap is presented in the form of monosilicic acid and is unloaded by Lsi6, a homolog of Lsi1 in rice. Lsi6 is also involved in the inter-vascular transfer of Si at the node, which is necessary for preferential Si distribution to the panicles.
doi:10.2183/pjab.87.377
PMCID: PMC3171283  PMID: 21785256
localization; distribution; uptake; silicon; transporter
19.  Phytoliths Analysis for the Discrimination of Foxtail Millet (Setaria italica) and Common Millet (Panicum miliaceum) 
PLoS ONE  2009;4(2):e4448.
Foxtail millet (Setaria italica) and Common millet (Panicum miliaceum) are the oldest domesticated dry farming crops in Eurasia. Identifying these two millets in the archaeobotanical remains are still problematic, especially because the millet grains preserve only when charred. Phytoliths analysis provides a viable method for identifying this important crop. However, to date, the identification of millet phytoliths has been questionable, because very little study has been done on their morphometry and taxonomy. Particularly, no clear diagnostic feature has been used to distinguish between Foxtail millet and Common millet. Here we examined the anatomy and silicon structure patterns in the glumes, lemmas, and paleas from the inflorescence bracts in 27 modern plants of Foxtail millet, Common millet, and closely related grasses, using light microscopy with phase-contrast and microscopic interferometer. Our research shows that five key diagnostic characteristics in phytolith morphology can be used to distinguish Foxtail millet from Common millet based on the presence of cross-shaped type, regularly arranged papillae, Ω-undulated type, endings structures of epidermal long cell, and surface ridgy line sculpture in the former species. We have established identification criteria that, when used together, give the only reliable way of distinguishing between Foxtail millet and Common millet species based on their phytoliths characteristics, thus making a methodological contribution to phytolith research. Our findings also have important implications in the fields of plant taxonomy, agricultural archaeology, and the culture history of ancient civilizations.
doi:10.1371/journal.pone.0004448
PMCID: PMC2636886  PMID: 19212442
20.  Interspecific Variation in Compensatory Regrowth to Herbivory Associated with Soil Nutrients in Three Ficus (Moraceae) Saplings 
PLoS ONE  2012;7(9):e45092.
Plant compensatory regrowth is an induced process that enhances plant tolerance to herbivory. Plant behavior against herbivores differs between species and depends on resource availability, thus making general predictions related to plant compensatory regrowth difficult. To understand how soil nutrients determine the degree of compensatory regrowth for different plant species, we selected saplings of three Ficus species and treated with herbivore insects and artificial injury in both glasshouse conditions and in the field at two soil nutrient levels. Compensatory regrowth was calculated by biomass, relative growth rate and photosynthetic characteristics. A similar pattern was found in both the glasshouse and in the field for species F. hispida, where overcompensatory regrowth was triggered only under fertile conditions, and full compensatory regrowth occurred under infertile conditions. For F. auriculata, overcompensatory regrowth was stimulated only under infertile conditions and full compensatory regrowth occurred under fertile conditions. Ficus racemosa displayed full compensatory regrowth in both soil nutrient levels, but without overcompensatory regrowth following any of the treatments. The three Ficus species differed in biomass allocation following herbivore damage and artificial injury. The root/shoot ratio of F. hispida decreased largely following herbivore damage and artificial injury, while the root/shoot ratio for F. auriculata increased against damage treatments. The increase of shoot and root size for F. hispida and F. auriculata, respectively, appeared to be caused by a significant increase in photosynthesis. The results indicated that shifts in biomass allocation and increased photosynthesis are two of the mechanisms underlying compensatory regrowth. Contrasting patterns among the three Ficus species suggest that further theoretical and empirical work is necessary to better understand the complexity of the plant responses to herbivore damage.
doi:10.1371/journal.pone.0045092
PMCID: PMC3440311  PMID: 22984616
21.  Effects of the Fungal Endophyte, Neotyphodium lolii, on Net Photosynthesis and Growth Rates of Perennial Ryegrass (Lolium perenne) are Independent of In Planta Endophyte Concentration 
Annals of Botany  2006;98(2):379-387.
• Background and Aims Neotyphodium lolii is a fungal endophyte of perennial ryegrass (Lolium perenne), improving grass fitness through production of bioactive alkaloids. Neotyphodium species can also affect growth and physiology of their host grasses (family Poaceae, sub-family Pooideae), but little is known about the mechanisms. This study examined the effect of N. lolii on net photosynthesis (Pn) and growth rates in ryegrass genotypes differing in endophyte concentration in all leaf tissues.
• Methods Plants from two ryegrass genotypes, Nui D and Nui UIV, infected with N. lolii (E+) differing approx. 2-fold in endophyte concentration or uninfected clones thereof (E−) were grown in a controlled environment. For each genotype × endophyte treatment, plant growth rates were assessed as tillering and leaf extension rates, and the light response of Pn, dark respiration and transpiration measured in leaves of young (30–45 d old) and old (>90 d old) plants with a single-chamber open infrared gas-exchange system.
• Key Results Neotyphodium lolii affected CO2-limited rates of Pn, which were approx. 17 % lower in E+ than E− plants (P < 0·05) in the young plants. Apparent photon yield and dark respiration were unaffected by the endophyte (P > 0·05). Neotyphodium lolii also decreased transpiration (P < 0·05), but only in complete darkness. There were no endophyte effects on Pn in the old plants (P > 0·05). E+ plants grew faster immediately after replanting (P < 0·05), but had approx. 10 % lower growth rates during mid-log growth (P < 0·05) than E− plants, but there was no effect on final plant biomass (P > 0·05). The endophyte effects on Pn and growth tended to be more pronounced in Nui UIV, despite having a lower endophyte concentration than Nui D.
• Conclusions Neotyphodium lolii affects CO2 fixation, but not light interception and photochemistry of Pn. The impact of N. lolii on plant growth and photosynthesis is independent of endophyte concentration in the plant, suggesting that the endophyte mycelium is not simply an energy drain to the plant. However, the endophyte effects on Pn and plant growth are strongly dependent on the plant growth phase.
doi:10.1093/aob/mcl108
PMCID: PMC2803460  PMID: 16735403
Lolium perenne; Neotyphodium lolii; perennial ryegrass; grass endophyte; net photosynthesis; tillering rate; leaf extension rate; plant fitness
22.  The ameliorative effect of silicon on soybean seedlings grown in potassium-deficient medium 
Annals of Botany  2010;105(6):967-973.
Background and Aims
Crop yield and nutritional quality are significantly reduced when potassium (K) in soil is deficient. As a beneficial element for plants, silicon (Si) is effective in alleviating the toxic effects of mineral nutrients. However, the roles played by Si in mediating deficiency in essential mineral nutrients in general and K in particular have not been investigated.
Methods
To evaluate the role of Si in K deficiency-induced inhibition of growth of soybean (Glycine max) seedlings, the effects of K deficiency on shoot and root growth, hydrogen peroxide accumulation, K contents, lipid peroxidation and activities of antioxidant enzymes in the absence and presence of 2 mm sodium silicate (Na2SiO3) were investigated.
Key Results
Both shoot and root biomass of soybean seedlings were markedly reduced when grown in K-deficient medium (1 mm K) compared with those grown in K-sufficient medium (5 mm). Addition of Na2SiO3 significantly ameliorated the K deficiency-induced reductions in shoot and root growth. Sodium silicate enhanced K concentrations in leaf, stem and root of K-deficient seedlings by 105·4, 83·4 and 58·8 %, respectively. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents in soybean seedlings were increased by 25 and 97 %, respectively, when exposed to K-deficient medium. These increases in accumulation of H2O2 and MDA were removed by addition of Na2SiO3. Addition of Na2SiO3 reduced the K deficiency-induced increases in activities of superoxide dismutase, catalase and peroxidase.
Conclusions
Application of Si to soybean seedlings grown in K-deficient medium markedly enhanced K use efficiency. Therefore, Si not only increases tolerance to nutrient toxicity, but also ameliorates symptoms associated with deficiency in essential nutrients in plants.
doi:10.1093/aob/mcq063
PMCID: PMC2876006  PMID: 20338952
Antioxidant enzyme activities; oxidative stress; potassium deficiency; silicon; soybean; Glycine max
23.  Earthworm and belowground competition effects on plant productivity in a plant diversity gradient 
Oecologia  2009;161(2):291-301.
Diversity is one major factor driving plant productivity in temperate grasslands. Although decomposers like earthworms are known to affect plant productivity, interacting effects of plant diversity and earthworms on plant productivity have been neglected in field studies. We investigated in the field the effects of earthworms on plant productivity, their interaction with plant species and functional group richness, and their effects on belowground plant competition. In the framework of the Jena Experiment we determined plant community productivity (in 2004 and 2007) and performance of two phytometer plant species [Centaurea jacea (herb) and Lolium perenne (grass); in 2007 and 2008] in a plant species (from one to 16) and functional group richness gradient (from one to four). We sampled earthworm subplots and subplots with decreased earthworm density and reduced aboveground competition of phytometer plants by removing the shoot biomass of the resident plant community. Earthworms increased total plant community productivity (+11%), legume shoot biomass (+35%) and shoot biomass of the phytometer C. jacea (+21%). Further, phytometer performance decreased, i.e. belowground competition increased, with increasing plant species and functional group richness. Although single plant functional groups benefited from higher earthworm numbers, the effects did not vary with plant species and functional group richness. The present study indicates that earthworms indeed affect the productivity of semi-natural grasslands irrespective of the diversity of the plant community. Belowground competition increased with increasing plant species diversity. However, belowground competition was modified by earthworms as reflected by increased productivity of the phytometer C. jacea. Moreover, particularly legumes benefited from earthworm presence. Considering also previous studies, we suggest that earthworms and legumes form a loose mutualistic relationship affecting essential ecosystem functions in temperate grasslands, in particular decomposition and plant productivity. Further, earthworms likely alter competitive interactions among plants and the structure of plant communities by beneficially affecting certain plant functional groups.
doi:10.1007/s00442-009-1374-1
PMCID: PMC2719079  PMID: 19526252
Aboveground–belowground interactions; Diversity–productivity relationship; Grassland; Positive interactions
24.  Above- and below-ground vertebrate herbivory may each favour a different subordinate species in an aquatic plant community 
Oecologia  2009;162(1):199-208.
At least two distinct trade-offs are thought to facilitate higher diversity in productive plant communities under herbivory. Higher investment in defence and enhanced colonization potential may both correlate with decreased competitive ability in plants. Herbivory may thus promote coexistence of plant species exhibiting divergent life history strategies. How different seasonally tied herbivore assemblages simultaneously affect plant community composition and diversity is, however, largely unknown. Two contrasting types of herbivory can be distinguished in the aquatic vegetation of the shallow lake Lauwersmeer. In summer, predominantly above-ground tissues are eaten, whereas in winter, waterfowl forage on below-ground plant propagules. In a 4-year exclosure study we experimentally separated above-ground herbivory by waterfowl and large fish in summer from below-ground herbivory by Bewick’s swans in winter. We measured the individual and combined effects of both herbivory periods on the composition of the three-species aquatic plant community. Herbivory effect sizes varied considerably from year to year. In 2 years herbivore exclusion in summer reinforced dominance of Potamogeton pectinatus with a concomitant decrease in Potamogeton pusillus, whereas no strong, unequivocal effect was observed in the other 2 years. Winter exclusion, on the other hand, had a negative effect on Zannichellia palustris, but the effect size differed considerably between years. We suggest that the colonization ability of Z. palustris may have enabled this species to be more abundant after reduction of P. pectinatus tuber densities by swans. Evenness decreased due to herbivore exclusion in summer. We conclude that seasonally tied above- and below-ground herbivory may each stimulate different components of a macrophyte community as they each favoured a different subordinate plant species.
doi:10.1007/s00442-009-1450-6
PMCID: PMC2776151  PMID: 19756762
Aquatic macrophytes; Waterfowl; Tubers; Competition colonization trade-off; Bare patch formation
25.  Herbivore induced plant volatiles 
Plant Signaling & Behavior  2011;6(12):1973-1978.
Plants respond to herbivory through different defensive mechanisms. The induction of volatile emission is one of the important and immediate response of plants to herbivory. Herbivore-induced plant volatiles (HIPVs) are involved in plant communication with natural enemies of the insect herbivores, neighboring plants, and different parts of the damaged plant. Release of a wide variety of HIPVs in response to herbivore damage and their role in plant-plant, plant-carnivore and intraplant communications represents a new facet of the complex interactions among different trophic levels. HIPVs are released from leaves, flowers, and fruits into the atmosphere or into the soil from roots in response to herbivore attack. Moreover, HIPVs act as feeding and/or oviposition deterrents to insect pests. HIPVs also mediate the interactions between the plants and the microorganisms. This review presents an overview of HIPVs emitted by plants, their role in plant defense against herbivores and their implications for pest management.
doi:10.4161/psb.6.12.18053
PMCID: PMC3337190  PMID: 22105032
herbivory; indirect defense; induced resistance; plant defense; volatiles

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