Effects of seed addition on beetle assemblages during the re-creation of species-rich lowland hay meadows

Species-rich lowland hay meadows are of conservation importance for both plants and invertebrates; however, they have declined in area across Europe as a result of conversion to other land uses and management intensification. The re-creation of these grasslands on ex-arable land provides a valuable approach to increasing the extent and conservation value of this threatened habitat. Over a 3-year period a replicated block design was used to test whether introducing seeds promoted the re-creation of both plant and phytophagous beetle assemblages typical of a target hay meadow. Seeds were harvested from local hay meadows, and applied to experimental plots in the form of either green hay or brush harvesting seeds. Green hay spreading achieved the greatest success in re-creating plant and phytophagous beetle assemblages. While re-creation success increased over time for both taxa, for the phytophagous beetles the greatest increase in re-creation success relative to the establishment year also occurred where green hay was applied. We also considered the phytophagous beetles in terms of functional traits that describe host plant specificity, larval feeding location and dispersal. Phytophagous beetle functional trait composition was most similar to the target hay meadow assemblage where some form of seed addition was used, i.e. hay spreading or brush harvested seeds. This study identified the importance of introducing target plant species as a mechanism to promote the re-creation of phytophagous beetle communities. Seed addition methods (e.g. green hay spreading) are crucial to successful hay meadow re-creation.

Hay strewing, brush harvesting of seed and soil disturbance as tools for the enhancement of botanical diversity in grasslands

Since the middle of the last century agricultural intensification within Europe has led to a drastic decline in the extent of botanically diverse grasslands. Whilst measures to enhance the diversity of agriculturally-improved grasslands are in place, success has often been limited. One of the primary factors limiting success is the paucity of sources of propagules of desirable species in the surrounding landscape. The restoration of two contrasting grassland types lowland hay meadow and chalk grassland) was examined using a replicated block experiment to assess the effectiveness of two methods of seed application (hay strewing and brush harvesting) and two methods of pre-treatment disturbance (power harrowing and turf stripping). The resulting changes in botanical composition were monitored for 4 years. Seed addition by both methods resulted in significant temporal trends in plant species composition and increases in plant species richness, which were further enhanced by disturbance. Power harrowing increased the effectiveness of the seed addition treatments at the lowland hay meadow site. At the chalk grassland site a more severe disturbance created by turf stripping was used and shown to be preferable. Whilst both hay strewing and brush harvesting increased plant species richness, hay strewing was more effective at creating a sward similar to that of the donor site. Soil disturbance and seed application rate at the recipient site and timing of the hay cut at the donor site are all factors to be considered prior to the commencement of restoration management. (c) 2006 Elsevier Ltd. All rights reserved.

Contrasting success in the restoration of plant and phytophagous beetle assemblages of species-rich mesotrophic grasslands

Over the last 60 years changes to the management of species-rich mesotrophic grasslands have resulted in the large-scale loss and degradation of this habitat across Europe. Restoration of such grasslands on agriculturally improved pastures provides a potentially valuable approach to the conservation of these threatened areas. Over a four-year period a replicated block design was used to test the effects of seed addition (green hay spreading and brush harvest collection) and soil disturbance on the restoration of phytophagous beetle and plant communities. Patterns of increasing restoration success, particularly where hay spreading and soil disturbance were used in combination, were identified for the phytophagous beetles. In the case of the plants, however, initial differences in restoration success in response to these same treatments were not followed by subsequent temporal changes in plant community similarity to target mesotrophic grassland. It is possible that the long-term consequences of the management treatments would not be the establishment of beetle and plant communities characteristic of the targets for restoration. Restoration management to enhance plant establishment using hay spreading and soil disturbance techniques would, however, still increase community similarity in both taxa to that of species-rich mesotrophic grasslands, and so raise their conservation value.

Limiting factors in the restoration of UK grassland beetle assemblages

Grasslands restoration is a key management tool contributing to the long-term maintenance of insect populations, providing functional connectivity and mitigating against extinction debt across landscapes. As knowledge of grassland insect communities is limited, the lag between the initiation of restoration and the ability of these new habitats to contribute to such processes is unclear. Using ten data sets, ranging from 3 to 14 years, we investigate the lag between restoration and the establishment of phytophagous beetle assemblages typical of species rich grasslands. We used traits and ecological characteristics to determine factors limiting beetle colonisation, and also considered how food-web structure changed during restoration. For sites where seed addition of host-plants occurred the success in replicating beetle assemblages increased over time following a negative exponential function. Extrapolation beyond the existing data set tentatively suggested that success would plateau after 20 years, representing a c. 60% increase in assemblage similarity to target grasslands. In the absence of seed addition, similarity to the target grasslands showed no increase over time. Where seed addition was used the connectance of plant-herbivore food webs decreased over time, approaching values typical of species rich grasslands after c. 7 years. This trend was, however, dependent on the inclusion of a single site containing data in excess of 6 years of restoration management. Beetles not capable of flight, those showing high degrees of host-plant specialisation and species feeding on nationally rare host plants take between 1 and 3 years longer to colonise. Successful grassland restoration is underpinned by the establishment of host-plants, although individual species traits compound the effects of poor host-plant establishment to slow colonisation. The use of pro-active grassland restoration to mitigate against future environmental change should account for lag periods in excess of 10 years if the value of these habitats is to be fully realised.

Microbial relatives of the seed storage proteins of higher plants: conservation of structure and diversification of function during evolution of the cupin superfamily.

This review summarizes the recent discovery of the cupin superfamily (from the Latin term "cupa," a small barrel) of functionally diverse proteins that initially were limited to several higher plant proteins such as seed storage proteins, germin (an oxalate oxidase), germin-like proteins, and auxin-binding protein. Knowledge of the three-dimensional structure of two vicilins, seed proteins with a characteristic beta-barrel core, led to the identification of a small number of conserved residues and thence to the discovery of several microbial proteins which share these key amino acids. In particular, there is a highly conserved pattern of two histidine-containing motifs with a varied intermotif spacing. This cupin signature is found as a central component of many microbial proteins including certain types of phosphomannose isomerase, polyketide synthase, epimerase, and dioxygenase. In addition, the signature has been identified within the N-terminal effector domain in a subgroup of bacterial AraC transcription factors. As well as these single-domain cupins, this survey has identified other classes of two-domain bicupins including bacterial gentisate 1, 2-dioxygenases and 1-hydroxy-2-naphthoate dioxygenases, fungal oxalate decarboxylases, and legume sucrose-binding proteins. Cupin evolution is discussed from the perspective of the structure-function relationships, using data from the genomes of several prokaryotes, especially Bacillus subtilis. Many of these functions involve aspects of sugar metabolism and cell wall synthesis and are concerned with responses to abiotic stress such as heat, desiccation, or starvation. Particular emphasis is also given to the oxalate-degrading enzymes from microbes, their biological significance, and their value in a range of medical and other applications.

Effects of desiccation on seed germination, cracking, fungal infection and survival in storage of Sterculia foetida L

Seeds of Sterculia foetida were tested for germination following desiccation and subsequent hermetic storage. Whereas seeds at 10.3% moisture content were intact and provided 98% germination, further desiccation reduced germination substantially. The majority of seed coats had cracked after desiccation to 5.1% moisture content. Ability to germinate was not reduced after 12 months' hermetic storage at 10.3% and 7.3% moisture content at 15 degrees C or -18 degrees C, but was reduced considerably at 5.1%. Fungal infection was detected consistently for cracked seeds in germination tests and they did not germinate. However, almost all embryos extracted from cracked seeds germinated if first disinfected with sodium hypochlorite (1%, 5 minutes). In addition. 80 -100% of disinfected extracted embryos from cracked seeds stored hermetically for 28 d at -18 degrees C or -82 degrees C with 3.3% to 6.0% moisture content, and excised embryos stored in this way, were able to germinate. Hence. failure of the very dry seeds of Sterculia foetida to germinate was not due to embryo death from desiccation but to cracking increasing susceptibility to fungal infection upon rehydration. Cracking was associated negatively and strongly with relative humidity and appears to be a mechanical consequence of substantial differences between the isotherms of whole seeds compared with cotyledons and axes.

Seed storage of Avicennia alba B1

Seeds of Avicennia alba BI. matured at high moisture content and were sensitive to desiccation: no seeds survived desiccation below 35% moisture content. The effect on survival of a factorial combination of five moist storage treatments (fresh seeds in a polyethylene bag, open with water sprayed over regularly, mixed with sand at 10% moisture content, mixed with moist paddy hulls, or naked seeds mixed with sand at 10% moisture content) and three temperatures (28-30degreesC, 17degreesC and 8-10degreesC) was investigated. In addition, seeds were mixed with sand at 5% moisture content and stored at 17degreesC in order to determine the effect of sand moisture content on seed moisture content and viability during storage. Avicennia alba showed recalcitrant seed storage behaviour, but 75% of the seeds remained viable after four months' moist (45-47% moisture content) storage in 10% moisture content sand at 17degreesC.

The impact of Solanum elaeagnifolium, an invasive plant in the Mediterranean, on the flower visitation and seed set of the native co-flowering species Glaucium flavum

We examined the effect of the invasive Solanum elaeagnifolium (Solanaceae) on flower visitation patterns and seed set of the co-flowering native Glaucium flavum (Papaveraceae). We observed flowering G. flavum plants in invaded and uninvaded sites and found that G. flavum flowers in uninvaded sites received significantly more total visits. In addition, we hand-pollinated flowers on plants of G. flavum with (i) pure conspecific pollen, (ii) pure S. elaeagnifolium pollen and (iii) three different mixtures of the two types of pollen (containing 25, 50 and 75% invasive pollen). As a control, flowers were left unmanipulated or were permanently bagged. Seed set did not differ significantly between flowers receiving pollen mixtures and pure conspecific pollen. However, in the open pollination treatment, seed set was significantly lower than in the 100% conspecific pollen treatment, which suggests pollen limitation. Bagged flowers had very low seed set. G. flavum was generally resilient against the deposition of S. elaeagnifolium pollen.

The role of the pod in seed development: strategies for manipulating yield

Pods play a key role in encapsulating the developing seeds and protecting them from pests and pathogens. In addition to this protective function, it has been shown that the photosynthetically active pod wall contributes assimilates and nutrients to fuel seed growth. Recent work has revealed that signals originating from the pod may also act to coordinate grain filling and regulate the reallocation of reserves from damaged seeds to those that have retained viability. In this review we consider the evidence that pods can regulate seed growth and maturation, particularly in members of the Brassicaceae family, and explore how the timing and duration of pod development might be manipulated to enhance either the quantity of crop yield or its nutritional properties.

Selected wheat seed defense proteins exhibit competitive binding to model microbial lipid interfaces

Puroindolines (Pins) and purothionins (Pths) are basic, amphiphilic, cysteine-rich wheat proteins that play a role in plant defense against microbial pathogens. We have examined the co-adsorption and sequential addition of Pins (Pin-a, Pin-b and a mutant form of Pin-b with Trp-44 to Arg-44 substitution) and β-purothionin (β-Pth) model anionic lipid layers, using a combination of surface pressure measurements, external reflection FTIR spectroscopy and neutron reflectometry. Results highlighted differences in the protein binding mechanisms, and in the competitive binding and penetration of lipid layers between respective Pins and β-Pth. Pin-a formed a blanket-like layer of protein below the lipid surface that resulted in the reduction or inhibition of β-Pth penetration of the lipid layer. Wild-type Pin-b participated in co-operative binding with β-Pth, whereas the mutant Pin-b did not bind to the lipid layer in the presence of β-Pth. The results provide further insight into the role of hydrophobic and cationic amino acid residues in antimicrobial activity.

Patterns of pollen and seed dispersal in a small, fragmented population of the wind-pollinated tree Araucaria angustifolia in southern Brazil

Paternity analysis based on eight microsatellite loci was used to investigate pollen and seed dispersal patterns of the dioecious wind- pollinated tree, Araucaria angustifolia. The study sites were a 5.4 ha isolated forest fragment and a small tree group situated 1.7 km away, located in Paran alpha State, Brazil. In the forest fragment, 121 males, 99 females, 66 seedlings and 92 juveniles were mapped and genotyped, together with 210 seeds. In the tree group, nine male and two female adults were mapped and genotyped, together with 20 seeds. Paternity analysis within the forest fragment indicated that at least 4% of the seeds, 3% of the seedlings and 7% of the juveniles were fertilized by pollen from trees in the adjacent group, and 6% of the seeds were fertilized by pollen from trees outside these stands. The average pollination distance within the forest fragment was 83 m; when the tree group was included the pollination distance was 2006m. The average number of effective pollen donors was estimated as 12.6. Mother- trees within the fragment could be assigned to all seedlings and juveniles, suggesting an absence of seed immigration. The distance of seedlings and juveniles from their assigned mother- trees ranged from 0.35 to 291m ( with an average of 83m). Significant spatial genetic structure among adult trees, seedlings, and juveniles was detected up to 50m, indicating seed dispersal over a short distance. The effective pollination neighborhood ranged from 0.4 to 3.3 ha. The results suggest that seed dispersal is restricted but that there is longdistance pollen dispersal between the forest fragment and the tree group; thus, the two stands of trees are not isolated.

Influence of sodium chloride on seed germination and seedling root growth of cotton (Gossypium hirsutum L.)

Response of cotton (Gossypium hirsutum L. cv. NIAB-78) to salinity, in terms of seed germination, seedling root growth and root Na+ and K+ content was determined in a laboratory experiment. Cotton seeds were exposed to increasing salinity levels using germination water with Sodium chloride concentrations of 0, 50, 100, 150 and 200 mM, to provide different degrees of salt stress. Germinated seeds were counted and roots were harvested at 24, 48, 72 and 96 h after the start of the experiment. It appeared that seed germination was only slightly affected by an increase in salinity (in most cases the differences between treatment were non-significant), whereas root length, root growth rate, root fresh and dry weights were severely affected, generally highly significant differences in these variables were found for comparisons involving most combinations of salinity levels, in particular with increased incubation period. K+ contents decreased with increasing salinity levels, although differences in K+ content were only significant when comparing the control and the 4 salinity levels. Na+ content of the roots increased with increasing levels of NaCl in the germination water, suggesting an exchange of K+ for Na+. The ratio K+/Na+ strongly decreased with rising levels of salinity from around 4.5 for the control to similar to 1 at 200 mM NaCl.

Enantioselective Intramolecular Michael Addition of Nitronates onto Conjugated Esters: Access to Cyclic γ-Amino Acids with up to Three Stereocenters

A highly stereoselective synthesis of conformationally constrained cyclic γ-amino acids has been devised. The key step involves an intramolecular cyclization of a nitronate onto a conjugated ester, promoted by a bifunctional thiourea catalyst. This methodology has been successfully applied to generate a variety of γ-amino acids, including some containing three contiguous stereocenters, with very high diastereoselectivity and excellent enantioselectivity. It is postulated that an interaction that is key to the success of the process is the simultaneous coordination of the thiourea functionality to both the conjugated ester and the nitronate. Finally, the synthetic utility of these compounds is demonstrated in the synthesis of two dipeptides derived from the C- and N-termini.

Effects of patch size and density on flower visitation and seed set of wild plants: a pan-European approach

1. Habitat fragmentation can affect pollinator and plant population structure in terms of species composition, abundance, area covered and density of flowering plants. This, in turn, may affect pollinator visitation frequency, pollen deposition, seed set and plant fitness. 2. A reduction in the quantity of flower visits can be coupled with a reduction in the quality of pollination service and hence the plants’ overall reproductive success and long-term survival. Understanding the relationship between plant population size and⁄ or isolation and pollination limitation is of fundamental importance for plant conservation. 3. Weexamined flower visitation and seed set of 10 different plant species fromfive European countries to investigate the general effects of plant populations size and density, both within (patch level) and between populations (population level), on seed set and pollination limitation. 4. Wefound evidence that the effects of area and density of flowering plant assemblages were generally more pronounced at the patch level than at the population level. We also found that patch and population level together influenced flower visitation and seed set, and the latter increased with increasing patch area and density, but this effect was only apparent in small populations. 5. Synthesis. By using an extensive pan-European data set on flower visitation and seed set we have identified a general pattern in the interplay between the attractiveness of flowering plant patches for pollinators and density dependence of flower visitation, and also a strong plant species-specific response to habitat fragmentation effects. This can guide efforts to conserve plant–pollinator interactions, ecosystem functioning and plant fitness in fragmented habitats.