Selection of trait combinations through bee and fly visitation to flowers of Polemonium foliosissimum

Pollinators are known to exert natural selection on floral traits, but the extent to which combinations of floral traits are subject to correlational selection (nonadditive effects of two traits on fitness) is not well understood. Over two years, we used phenotypic manipulations of plant traits to test for effects of flower colour, flower shape and their interaction on rates of pollinator visitation to Polemonium foliosissimum. We also tested for correlational selection based on weighting visitation by the amount of conspecific pollen delivered per visit by each category of insect visitor. Although bumblebees were the presumed pollinators, solitary bees and flies contributed substantially (42%) to pollination. In manipulations of one trait at a time, insects visited flowers presenting the natural colour and shape over flowers manipulated to present artificial mutants with either paler colour or a more open or more tubular flower. When both colour and shape were manipulated in combination, selection on both traits arose, with bumblebees responding mainly to colour and flies responding mainly to shape. Despite selection on both floral traits, in a year with many bumblebees, we saw no evidence for correlational selection of these traits. In a year when flies predominated, fly visitation showed a pattern of correlational selection, but not favouring the natural phenotype, and correlational selection was still not detected for expected pollen receipt. These results show that flower colour and shape are subject to pollinator‐mediated selection and that correlational selection can be generated based on pollinator visitation alone, but provide no evidence for correlational selection specifically for the current phenotype.     

Specialty oilseed crops provide an abundant source of pollen for pollinators and beneficial insects

The continuing pollinator crisis is due, in part, to the lack of year‐round floral resources. In intensive farming regions, such as the Upper Midwest (UMW) of the USA, natural and pastoral vegetation largely has been replaced by annual crops such as maize (Zea mays L.), soyabean (Glycine max L.) and wheat (Triticum spp.). Neither the energy (nectar) nor protein (pollen) needs of pollinating and other beneficial insects are being met sufficiently by the new, high‐intensity, agricultural landscape. Several potentially useful oilseed crops can be grown in the UMW, and many of these oilseeds are highly attractive to beneficial insects. Prior research showed that some of these oilseeds produced abundant nectar, but their corresponding values for pollen production are unknown. Accordingly, the aim of our research was to document pollen (and protein) production per unit area of twelve oilseed crops grown in Minnesota and associate these values with levels of beneficial insect visitation during anthesis. Our results show that oilseed crops such as camelina (Camelina sativa L.), flax (Linum usitatissimum L.) and pennycress (Thlaspi arvense L.) produce relatively little pollen (≤40 kg/ha); borage (Borago officinalis L.), calendula (Calendula officinalis L.), canola (Brassica napus L.), crambe (Crambe abyssianica Hochst) and cuphea (Cuphea viscosissima Jacq. × Cuphea lanceolata W. T. Aiton) produce bountiful pollen resources (50–150 kg/ha); and oilseed echium (Echium plantagineum L.) generates massive amounts of pollen (>400 kg/ha), about 50% of which is protein. Our study is unique in presenting a season‐long perspective of pollen production in alternative oilseed crops, a resource valuable to pollen‐feeding insects such as managed and wild bees. Diversification of UMW landscapes that includes alternative oilseed crops such as oilseed echium and cuphea can potentially provide a ready source of pollen and protein to help combat pollinator decline.     

Patch size has no effect on insect visitation rate per unit area in garden-scale flower patches

Previous studies investigating the effect of flower patch size on insect flower visitation rate have compared relatively large patches (10–1000s m²) and have generally found a negative relationship per unit area or per flower. Here, we investigate the effects of patch size on insect visitation in patches of smaller area (range c. 0.1–3.1 m²), which are of particular relevance to ornamental flower beds in parks and gardens. We studied two common garden plant species in full bloom with 6 patch sizes each: borage (Borago officinalis) and lavender (Lavandula × intermedia ‘Grosso’). We quantified flower visitation by insects by making repeated counts of the insects foraging at each patch. On borage, all insects were honey bees (Apis mellifera, n = 5506 counts). On lavender, insects (n = 737 counts) were bumble bees (Bombus spp., 76.9%), flies (Diptera, 22.4%), and butterflies (Lepidoptera, 0.7%). On both plant species we found positive linear effects of patch size on insect numbers. However, there was no effect of patch size on the number of insects per unit area or per flower and, on lavender, for all insects combined or only bumble bees. The results show that it is possible to make unbiased comparisons of the attractiveness of plant species or varieties to flower-visiting insects using patches of different size within the small scale range studied and make possible projects aimed at comparing ornamental plant varieties using existing garden flower patches of variable area.     

Pollinators and pollination of oilseed rape crops (Brassica napus L.) in Ireland: ecological and economic incentives for pollinator conservation

Pollinators are beneficial for many wild and crop plants. As a mass-flowering crop, oilseed rape has received much focus in terms of its pollination requirements but despite a threefold increase in area of cultivation of this crop in Ireland over the past 5 years, little is known about its pollination here. We surveyed the flower visiting insects found in commercial winter oilseed rape fields and evaluated the importance of different pollinator groups, investigated the contribution of insect pollination to oilseed rape seed production, and estimated the economic value of insect pollination to the crop at a national level. Our data showed that winter oilseed rape is visited by a wide variety of insect species, including the honeybee, bumblebees, solitary bees, and hoverflies. The honeybee, Eristalis hoverflies and bumblebees (especially Bombus sensu stricto and B. lapidarius) were the best pollinators of winter oilseed rape based on the number of pollen grains they carry, visitation rates per flower and their relative abundance per field. Exclusion of pollinators resulted in a 27 % decrease in the number of seeds produced, and a 30 % decrease in seed weight per pod in winter crops, with comparable values from a spring oilseed rape field also. The economic value of insect pollination to winter oilseed rape was estimated as €2.6 million per annum, while the contribution to spring oilseed rape was €1.3 million, resulting in an overall value of €3.9 million per annum. We can suggest the appropriate conservation and management of both honeybees and wild pollinators in agricultural areas to ensure continued provision of pollination services to oilseed rape, as a decrease in insect numbers has the potential to negatively influence crop yields.     

Pollination of Euphorbia dendroides by lizards and insects: Spatio-temporal variation in patterns of flower visitation

The patterns of flower visitation by lizards (Podarcis lilfordi, Lacertidae) and insects (mainly flies, bees and wasps) on the shrub Euphorbia dendroides, were studied in the island of Cabrera (Balearic Islands) during the flowering seasons of 1995 and 1996. Lizards act as true pollinators of the plant, moving large quantities of pollen within and among shrubs. To our knowledge, this is the first time that pollination by lizards has been empirically demonstrated. Variation in the quantitative component of pollination (frequency of visits × flower visitation rate) by the two groups of pollinators (lizards and insects) is documented at both spatial (within a plant population) and temporal scales (throughout the flowering season and between seasons). Variation in lizard density on a small spatial scale (within c. 200 m), presumably due to differences in vegetation cover, strongly affected their frequency of flower visitation. Insects were rather scarce, mainly because the plant flowers at a time (mid-March) when temperatures are still low. At the site where lizards were abundant, their frequency of flower visits was more than 3 times that of insects, they stayed on the shrubs about 3 times longer and visited about 8 times more cyathia per minute than did insects. Fruit and seed set were greater at this site, and this is attributed to the different frequency of flower visits by lizards, as shrubs are similar in size and produce similar amounts of cyathia in the two sites compared. Both, lizards and insects went more frequently to plants with large flower crops. However, flower crop was not associated with seed viability. We found no evidence for pollinator-mediated selection on plant traits related to fitness. Received: 28 August 1996 / Accepted: 26 February 1997     

Molecular tools enabling pennycress (Thlaspi arvense) as a model plant and oilseed cash cover crop

Thlapsi arvense L. (pennycress) is being developed as a profitable oilseed cover crop for the winter fallow period throughout the temperate regions of the world, controlling soil erosion and nutrients run‐off on otherwise barren farmland. We demonstrate that pennycress can serve as a user‐friendly model system akin to Arabidopsis that is well‐suited for both laboratory and field experimentation. We sequenced the diploid genome of the spring‐type Spring 32‐10 inbred line (1C DNA content of 539 Mb; 2n = 14), identifying variation that may explain phenotypic differences with winter‐type pennycress, as well as predominantly a one‐to‐one correspondence with Arabidopsis genes, which makes translational research straightforward. We developed an Agrobacterium‐mediated floral dip transformation method (0.5% transformation efficiency) and introduced CRISPR‐Cas9 constructs to produce indel mutations in the putative FATTY ACID ELONGATION1 (FAE1) gene, thereby abolishing erucic acid production and creating an edible seed oil comparable to that of canola. We also stably transformed pennycress with the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene, producing low‐viscosity acetyl‐triacylglycerol‐containing seed oil suitable as a diesel‐engine drop‐in fuel. Adoption of pennycress as a model system will accelerate oilseed‐crop translational research and facilitate pennycress’ rapid domestication to meet the growing sustainable food and fuel demands.     

Winter oilseed production for biofuel in the US Corn Belt: opportunities and limitations

Interest from the US commercial aviation industry and commitments established by the US Navy and Air Force to use renewable fuels has spurred interest in identifying and developing crops for renewable aviation fuel. Concern regarding greenhouse gas emissions associated with land‐use change and shifting land grown for food to feedstock production for fuel has encouraged the concept of intensifying current prominent cropping systems through various double cropping strategies. Camelina (Camelina sativa L.) and field pennycress (Thlaspi arvense L.) are two winter oilseed crops that could potentially be integrated into the corn (Zea mays L.)–soybean [(Glycine max (L.) Merr.] cropping system, which is the prominent cropping system in the US Corn Belt. In addition to providing a feedstock for renewable aviation fuel production, integrating these crops into corn–soybean cropping systems could also potentially provide a range of ecosystem services. Some of these include soil protection from wind and water erosion, soil organic C (SOC) sequestration, water quality improvement through nitrate reduction, and a food source for pollinators. However, integration of these crops into corn–soybean cropping systems also carries possible limitations, such as potential yield reductions of the subsequent soybean crop. This review identifies and discusses some of the key benefits and constraints of integrating camelina or field pennycress into corn–soybean cropping systems and identifies generalized areas for potential adoption in the US Corn Belt.     

Reconstructing the pollinator community and predicting seed set from hydrocarbon footprints on flowers

The measurement of insect visits to flowers is essential for basic and applied pollination ecology, but is often fraught with difficulty. Floral visitation is highly variable and observational studies are limited in scope due to the considerable time needed to acquire reliable data. Our study investigates whether the analysis of hydrocarbon residues (footprints) deposited by insects during flower visits allows the reconstruction of the visitor community and the prediction of seed set for large numbers of plants. In three consecutive years we recorded bumblebee visitation to wild plants of comfrey, Symphytum officinale, and later used gas chromatography/mass spectrometry (GC/MS) to quantify bumblebee-derived unsaturated hydrocarbons (UHCs) extracted from flowers. The UHCs washed from corollae were most similar to the tarsal UHC profile of the most abundant bumblebee species, Bombus pascuorum, in all 3 years. The species compositions of the bumblebee communities estimated from UHCs on flowers were also similar to those actually observed. There was a significant positive correlation between the observed number of visits by each of three bumblebee species (contributing 3–68% of the flower visits) and the estimated number of visits based on UHC profiles. Furthermore, significant correlations were obtained separately for workers and drones of two species. Seed set of comfrey plants was positively correlated with overall bumblebee visitation and the total amount of UHCs on flowers, suggesting the potential for pollen limitation. We suggest that quantifying cumulative footprint hydrocarbons provides a novel way to assess floral visitation by insects, and that this method can be used to predict seed set in pollen-limited plants.     

Pollination niche separation in a winter annual community

Summary Differences in major selective forces important in early and late successional communities should influence niche breadth and degree of overlap. Early successional species may not experience consistent, strong selection against competition and can be expected to have broader niches with more overlap than later successional species. This paper presents data from 2 early successional winter annuals that show clear niche separation under conditions in which coevolution is unlikely. Lactuca scariola L. and Erigeron annuus (L.) Pers. flower in mid-summer and attract the same insect visitors. Erigeron, the more common species, begins to flower in mid-morning when insect visitors are quite active. Lactuca flowers open earlier in the morning and close just before Erigeron flowers open. The early morning hours are not optimum for Lactuca visitation, as most of the visits occur just before floral closing. Both species can selffertilize without a vector, and Lactuca was introduced from Europe. It is, therefore, not possible that niche separation inflowering time and subsequent time of visit in Erigeron and Lactuca is a result of coevolutionary niche differentiation between the two species. This separation is likely to be a preadaptation resulting from coevolution with other species.     

Fruit set in perennial vetch (Vicia unijuga): reproductive system and insect role in pollination

Perennial vetch (Vicia unijuga) is a wild plant found in parts of East Asia and potentially valuable as a forage species for more extreme environments. Information on its reproductive system and pollination biology is needed for progress in domestication of the species. We characterized the reproductive system of perennial vetch as facultative xenogamy (i.e. it is largely cross‐pollinated by insects but is also self‐compatible and can self‐pollinate). There was no significant difference (P > 0.05) in the fruit set ratio between insect cross‐pollination and artificial cross‐pollination at the seed maturation stage, but natural self‐pollination was inefficient. In our study conducted on the Tibetan plateau, eight insect species (especially certain bumble bees) are identified as potential pollinators, and four other insect species belonging to the Lepidoptera and Diptera visited flowers but are unlikely to be pollinators. The flower visitation rate of wild bumble bees was 1.6–3.3 times higher than domestic honeybees, with Bombus lepidus having the highest visitation rate of 15.7 florets/min. The diurnal floret opening rhythm of perennial vetch was synchronized with diurnal activity of potential pollinators. Optimal pollination of perennial vetch would likely be achieved using wild bees, as they have behaviour characteristics and flower tripping ability necessary, and are present in sufficient numbers to be efficient pollinators of this crop. However, even with sufficiency of pollination, there remains a fundamental problem with low fruit set which requires further investigation from a plant biology perspective.     

Do artificial nectar feeders affect bat–plant interactions in an Ecuadorian cloud forest?

Plant–pollinator interactions are critical to ecosystems. However, when artificial nectar feeders are available in an area, they could draw pollinators away from plants. We tested the effects of artificial nectar feeders in an Ecuadorian cloud forest on four aspects of bat–plant interactions: (1) bat relative abundance; (2) bat pollen loads; (3) flower visitation rates, and (4) breeding success of a bat‐pollinated species (Burmeistera glabrata). We divided the study site into areas close to (~30 m) and far from (~500 m) three different feeder sites. At each distance, we captured nectar bats (Anoura caudifer, Anoura cultrata, and Lonchophylla robusta) to estimate their relative abundance and to collect pollen from fur and fecal samples. We also videotaped flowers to estimate bat visitation rates and recorded different breeding success variables of B. glabrata. We found that areas close to feeders have higher relative bat abundance by a factor of 40. In spite of this, the presence of feeders did not affect bat pollen loads, nor the flower visitation rates and breeding success of B. glabrata. Interestingly, there were differences in pollen loads between the three bat species, in that L. robusta individuals rarely carried pollen and were only captured near feeders.     

A draft genome of field pennycress (Thlaspi arvense) provides tools for the domestication of a new winter biofuel crop

Field pennycress (Thlaspi arvense L.) is being domesticated as a new winter cover crop and biofuel species for the Midwestern United States that can be double-cropped between corn and soybeans. A genome sequence will enable the use of new technologies to make improvements in pennycress. To generate a draft genome, a hybrid sequencing approach was used to generate 47 Gb of DNA sequencing reads from both the Illumina and PacBio platforms. These reads were used to assemble 6,768 genomic scaffolds. The draft genome was annotated using the MAKER pipeline, which identified 27,390 predicted protein-coding genes, with almost all of these predicted peptides having significant sequence similarity to Arabidopsis proteins. A comprehensive analysis of pennycress gene homologues involved in glucosinolate biosynthesis, metabolism, and transport pathways revealed high sequence conservation compared with other Brassicaceae species, and helps validate the assembly of the pennycress gene space in this draft genome. Additional comparative genomic analyses indicate that the knowledge gained from years of basic Brassicaceae research will serve as a powerful tool for identifying gene targets whose manipulation can be predicted to result in improvements for pennycress.     

The effects of floral display on pollinator visitation vary among populations ofPhacelia linearis (Hydrophyllaceae)

Summary Individual plants in gynodioecious populations ofPhacelia linearis (Hydrophyllaceae) vary in flower gender, flower size, and flower number. This paper reports the effects of variation in floral display on the visitation behaviour of this species' pollinators (mainly pollen-collecting solitary bees) in several natural and three experimental plant populations, and discusses the results in terms of the consequences for plant fitness. The working hypotheses were: (1) that because female plants do not produce pollen, pollen-collecting insects would visit hermaphrodite plants at a higher rate than female plants and would visit more flowers per hermaphrodite than per female; and (2) that pollinator arrival rate would increase with flower size and flower number, the two main components of visual display. These hypotheses were generally supported, but the effects of floral display on pollinator visitation varied substantially among plant populations. Hermaphrodites received significantly higher rates of pollinator arrivals and significantly higher rates of visits to flowers than did females in all experimental populations. Flower size affected arrival rate and flower visit rate positively in natural populations and in two of the three experimental populations. The flower size effect was significant only among female plants in one experimental population, and only among hermaphrodites in another. The effect of flower number on arrival rate was positive and highly significant in natural populations and in all experimental populations. In two out of three experimental populations, insects visited significantly more flowers per hermaphrodite than per female and visited more flowers on many-flowered plants than on few-flowered plants, but neither effect was detected in the third experimental population. Because seed production is not pollen-limited in this species, variation in pollinator visitation behaviour should mainly affect the male reproductive success of hermaphrodite plants. These findings suggest that pollinator-mediated natural selection for floral display inP. linearis varies in space and time.     

南京地区外来植物一年蓬上访花昆虫的多样性及其访花选择性的影响因素分析

为揭示外来植物一年蓬Erigeron annuus上的本土访花昆虫多样性和影响访花行为的因素, 本研究在南京郊区进行了连续2年的野外调查, 采用跨栏模型分析了环境因素如何影响昆虫的访花选择性, 即接受概率（测度是否接受一年蓬花）和访问频数（测度接受一年蓬花的程度）。调查发现, 访问一年蓬花的昆虫共计9目54科145种, 其中, 科丰富度占优势的是膜翅目、 鳞翅目和鞘翅目（均占总科数的20.75%）, 其次是双翅目（18.87%）和半翅目（13.21%）; 物种丰富度占优势的是双翅目（26.39%）, 其次是膜翅目（18.75%）、 半翅目（18.75%）、 鞘翅目（17.36%）和鳞翅目（15.38%）。多数目的物种丰富度在6-7月最高, 9月最低, 仅双翅目（食蚜蝇为主）在5月最高。运用跨栏模型对物种优势度最大的半翅目、 膜翅目和双翅目等的访花个体数量及其影响因素的分析结果表明： 影响半翅目和膜翅目对一年蓬花访问倾向（接受概率）的因素多于影响其访问频数的因素, 由此预测这些访花昆虫可能参考较多因素做出是否接受一年蓬花, 而依据较少线索做出访问程度的访花行为决策; 一年蓬植株密度影响半翅目和膜翅目昆虫的接受概率, 而花密度影响半翅目和双翅目昆虫的接受概率和访问频数, 说明靶标植物花的特性可能对访花昆虫的访花行为决策起主要作用。     

Managed bumble bees increase flower visitation but not fruit weight in polytunnel strawberry crops

Animal-mediated pollination is essential for the production and quality of fruits and seeds of many crops consumed by humans. However, crop pollination services might be compromised when wild pollinators are scarce. Managed pollinators are commonly used in crops to supplement such services with the assumption that they will enhance crop yield. However, information on the spatiotemporal pollinator-dependence of crops is still limited. We assessed the contribution of commercial bumble bee colonies compared to the available pollinator community on strawberry (‘Fortuna’ variety) flower visitation and strawberry quality across a landscape gradient of agricultural intensification (i.e. polytunnel berry crop cover). We used colonies of bumble bees in winter and in spring, i.e. when few and most wild pollinators are in their flight period, respectively. The placement of colonies increased visits of bumble bees to strawberry flowers, especially in winter. The use of bumble bee colonies did not affect flower visitation by other insects, mainly honey bees, hoverflies and other Diptera. Flower visitation by both honey bees and wild insects did not vary between seasons and was unrelated to the landscape gradient of berry crop cover. Strawberries were of the highest quality (i.e. weight) when insect-mediated pollination was allowed, and their quality was positively related to wild flower visitors in winter but not in spring. However, increased visits to strawberry flowers by managed bumble bees and honey bees had no effect on strawberry weight. Our results suggest that the pollination services producing high quality strawberry fruits are provided by the flower visitor community present in the study region without the need to use managed bumble bees.     

Manipulating field margins to increase predation intensity in fields of winter wheat (Triticum aestivum)

The effectiveness of natural enemies to control pests can be enhanced through habitat manipulation. However, due to the differences in their ecology, generalist and specialist species may respond differently to the same manipulation. Moreover, interactions among natural enemies (i.e. cannibalism, intraguild predation, hyperparasitism) may complicate the assumption that a higher density of natural enemies would increase the level of biological control. We investigated the natural enemy guild composition and the predation rate along flower vs. grass margins at the edge of winter wheat (Triticum aestivum) fields in Denmark. Natural enemies were sampled by pitfall trapping and by suction sampling; predation intensity was measured using two different sentinel prey methods: artificial caterpillars made of plasticine, and sentinel aphid colonies. Specialist and generalist species responded differently to the two margin types: specialists (mostly parasitic wasps) were attracted by the flower margins, while generalists (ground beetles, rove beetles and spiders) were more active in grass margins. The number of artificial caterpillars attacked was significantly greater in grass margins (mean = 48.9%, SD = 24.3) than in flower margins (mean = 30.7%, SD = 17.4). We found a significant positive relationship between the number of artificial caterpillars attacked by chewing insects, and activity density for large (≥15 mm) ground beetles. Predation of sentinel aphids in wheat fields did not vary significantly in relation to margin type. Our results suggest that flowering margins may be beneficial for canopy‐active specialist natural enemies, but grassy margins are more useful for ground‐active generalist predators.     

Visitation rate of pollinators and nectar robbers to the flowers and inflorescences of Tabebuia aurea (Bignoniaceae): effects of floral display size and habitat fragmentation

Large floral displays favour pollinator attraction and the import and export of pollen. However, large floral displays also have negative effects, such as increased geitonogamy, pollen discounting and nectar/pollen robber attraction. The size of the floral display can be measured at different scales (e.g. the flower, inflorescence or entire plant) and variations in one of these scales may affect the behaviour of flower visitors in different ways. Moreover, the fragmentation of natural forests may affect flower visitation rates and flower visitor behaviour. In the present study, video recordings of the inflorescences of a tree species (Tabebuia aurea) from the tropical savannah of central Brazil were used to examine the effect of floral display size at the inflorescence and tree scales on the visitation rate of pollinators and nectar robbers to the inflorescence, the number of flowers approached per visit, the number of visits per flower of potential pollinators and nectar robbers, and the interaction of these variables with the degree of landscape disturbance. Nectar production was quantified with respect to flower age. Although large bees are responsible for most of the pollination, a great diversity of flower insects visit the inflorescences of T. aurea. Other bee and hummingbird species are highly active nectar robbers. Increases in inflorescence size increase the visitation rate of pollinators to inflorescences, whereas increases in the number of inflorescences on the tree decrease visitation rates to inflorescences and flowers. This effect has been strongly correlated with urban environments in which trees with the largest floral displays are observed. Pollinating bees (and nectar robbers) visit few flowers per inflorescence and concentrate visits to a fraction of available flowers, generating an overdispersed distribution of the number of visits per inflorescence and per flower. This behaviour reflects preferential visits to young flowers (including flower buds) with a greater nectar supply.     

Pollinator-mediated interactions between a pathogenic fungus, Uromyces pisi (Pucciniaceae), and its host plant,Euphorbia cyparissias (Euphorbiaceae)

The plant Euphorbia cyparissias is commonly infected by rust fungi of the species complex Uromyces pisi. When infected, E. cyparissias is unable to flower, but instead is induced by the fungus to form pseudoflowers. Pseudoflowers are rosettes of yellow leaves upon which the fungus presents its gametes in a sweet-smelling fungal nectar. We hypothesized that the fungi, as they are heterothallic, are dependent on insect visitation to cross-fertilize their mating types. We confirmed that insects are required with an insect exclusion experiment. We further hypothesized that pseudoflowers of U. pisi interact with uninfected true host flowers through insects during their period of co-"flowering" in early spring. We conducted artificial array experiments in the field to test whether the two species share insects and whether they influenced each other's insect visitation. Insects moved between true flowers and pseudoflowers, but true flowers received more visits over all. Pseudoflowers and true flowers did not influence each other's visitation rates in mixtures. However, shorter visits were observed on pseudoflowers in mixtures than monocultures, suggesting that true flowers might be competitors for pseudoflowers. Further experiments are needed to determine whether the similarity of pseudoflowers to true flowers is adaptive.     

Impact of a soil sampling strategy on the spatial distribution and diversity of arbuscular mycorrhizal communities at a small scale in two winter cover crop rotational systems

It is vital to evaluate how soil sampling affects the spatial distribution and diversity of arbuscular mycorrhizal fungi (AMF) in agricultural ecosystems. Here, the impact of soil sampling on the spatial distribution and diversity of AMF in agricultural ecosystems was evaluated using field experiments. A molecular approach was taken to assess the spatial distribution of the AMF community from 1 m2 plots from fields cropped with soybeans [Glycine max (L.) Merr.], in rotation with wheat (Triticum aestivum L.) and winter fallow. Phylogenetic analysis revealed 18 AMF phylotypes, including five Glomus, three Gigaspora, two each from Racocetra, Acaulospora, and Funneliformis and one each from Claroideoglomus, Rhizophagus, Diversispora, and Paraglomus at different sampling points in this study. Our results showed that the molecular diversity and composition of AMF communities in soil cropped with wheat, or left fallow did not change at scales of < 1 m2. Canonical correspondence analysis (CCA) demonstrated that AMF communities by soil sampling point within each rotation were not significantly different. Thus, random soil sampling did not show any difference in AMF communities in soil under winter cover crop rotational systems, no matter where soil samples were collected from at a small scale (< 1 m2) in agricultural fields. Our results suggest that agricultural management can affect the diversity and composition of AMF communities more than soil sampling strategies.