Metabolomic analysis of pollen from honey bee hives and from canola flowers.

BACKGROUND AND AIMS: Pollen is essential for successful plant reproduction and critical for plant-pollinator mutualisms, as pollen is essential larval nutrition. However, we understand very little about the chemical constituents of pollen leading us to this exploratory study characterizing plant and beehive pollen. METHODS: We performed a metabolomics assay of canola flower pollen and beehive pollen. RESULTS AND DISCUSSION: The metabolome of canola pollen is affected by irrigation showing differences in lipids and non-polar secondary metabolites. Metabolome of beehive pollen is affected by plant source showing differences in pentose sugars, myo-inositol and furanose. Further research is needed to document the nutritional bases of plant-pollinator mutualism.

Floral longevity and autonomous selfing are altered by pollination and water availability in Collinsia heterophylla.

BACKGROUND AND AIMS: A plant investing in reproduction partitions resources between flowering and seed production. Under resource limitation, altered allocations may result in floral trait variations, leading to compromised fecundity. Floral longevity and timing of selfing are often the traits most likely to be affected. The duration of corolla retention determines whether fecundity results from outcrossing or by delayed selfing-mediated reproductive assurance. In this study, the role of pollination schedules and soil water availability on floral longevity and seed production is tested in Collinsia heterophylla (Plantaginaceae). METHODS: Using three different watering regimes and pollination schedules, effects on floral longevity and seed production were studied in this protandrous, flowering annual. KEY RESULTS: The results reveal that soil water status and pollination together influence floral longevity with low soil water and hand-pollinations early in the floral lifespan reducing longevity. However, early pollinations under excess water did not extend longevity, implying that resource surplus does not lengthen the outcrossing period. The results also indicate that pollen receipt, a reliable cue for fecundity, accelerates flower drop. Early corolla abscission under drought stress could potentially exacerbate sexual conflict in this protandrous, hermaphroditic species by ensuring self-pollen paternity and enabling male control of floral longevity. While pollination schedules did not affect fecundity, water stress reduced per-capita seed numbers. Unmanipulated flowers underwent delayed autonomous selfing, producing very few seeds, suggesting that inbreeding depression may limit benefits of selfing. CONCLUSIONS: In plants where herkogamy and dichogamy facilitate outcrossing, floral longevity determines reproductive success and mating system. Reduction in longevity under drought suggests a strong environmental effect that could potentially alter the preferred breeding mode in this mixed-mated species. Extrapolating the findings to unpredictable global drought cycles, it is suggested that in addition to reducing yield, water stress may influence the evolutionary trajectory of plant mating system.

Seasonal patterns of beneficial phytochemical availability in honey and stored pollen from honey bee colonies in large apiaries.

Honey bees (Apis mellifera L.; Hymenoptera, Apidae) are the most efficient pollinators in agroecosystems, responsible for the successful production of fruits, nuts, and vegetables, but they continue to face debilitating challenges. One of the major factors leading to these challenges could be linked to poor nutrition that results in weakening the colony, increasing susceptibility to pests and pathogens, and reducing the ability of bees to adapt to other abiotic stresses. Extensively used for commercial pollination, honey bee colonies regularly face exposure to limited diversity in their pollen diet as they are placed in flowering monocrops. Lack of access to diverse plant species compromises the availability of plant secondary compounds (phytochemicals), which, in small amounts, provide significant benefits to honey bee health. We analyzed the beneficial phytochemical content of honey and stored pollen (bee bread) samples from colonies in large apiaries through the active bee season. Samples were evaluated for 4 beneficial phytochemicals (caffeine, kaempferol, gallic acid, and p-coumaric acid), which have previously been shown to improve honey bee health. Our results, as relevant to the apiary locations in the study, indicated that p-coumaric acid is uniformly available throughout the season. Caffeine is completely absent, and gallic acid and kaempferol are not regularly available. Our results suggest the need to explore the potential to deliver beneficial phytochemicals as nutritional supplements to improve bee health. It may be vital for the pollination industry to consider such targeted dietary supplementation as beekeepers strive to meet the increasing demand for crop pollination services.

Dietary phytochemicals alter hypopharyngeal gland size in honey bee (Apis mellifera L.) workers.

Honey bees are the most efficient pollinators of several important fruits, nuts and vegetables and are indispensable for the profitable production of these crops. Health and performance of honey bee colonies have been declining for decades due to a combination of factors including poor nutrition, agrochemicals, pests and diseases. Bees depend on a diversity of plants for nutrition as pollen is the predominant protein and lipid source, and nectar, the source of carbohydrates for larval development. Additionally, pollen and nectar also contain small amounts of plant secondary metabolites or phytochemicals that are primarily plant defense compounds. Bees have coevolved to benefit from these compounds as seen by the improved longevity, pathogen tolerance and gut microbiome abundance in worker bees whose diets were supplemented with select phytochemicals. Here we investigate the impact of four phytochemicals, known to benefit bees, - caffeine, kaempferol, gallic acid and p-coumaric acid, on hypopharyngeal gland (HPG) size of nurse bees. Newly emerged bees were provided with 25 ppm of each of the four phytochemicals in 20% (w/v) sucrose solution and the size of HPGs were measured after a 10 d period. Bees that received p-coumaric acid or kaempferol showed a significant increase in HPG size. A significant decrease in HPG size was seen in bees receiving caffeine or gallic acid. The implication of our findings on worker bee ontogeny, transitioning from nurses to foragers and relevance to foraging related competencies are discussed. It is critical that bees have access to phytochemicals to ensure colony health and performance. Such access could be through natural habitats that provide a diversity of pollen and nectar sources or through dietary supplements for bee colonies.

Selenium accumulation in flowers and its effects on pollination.

• Selenium (Se) hyperaccumulation has a profound effect on plant-arthropod interactions. Here, we investigated floral Se distribution and speciation in flowers and the effects of floral Se on pollen quality and plant-pollinator interactions. • Floral Se distribution and speciation were compared in Stanleya pinnata, an Se hyperaccumulator, and Brassica juncea, a comparable nonhyperaccumulator. Pollen germination was measured from plants grown with varying concentrations of Se and floral visitation was compared between plants with high and low Se. • Stanleya pinnata preferentially allocated Se to flowers, as nontoxic methyl-selenocysteine (MeSeCys). Brassica juncea had higher Se concentrations in leaves than flowers, and a lower fraction of MeSeCys. For B. juncea, high floral Se concentration impaired pollen germination; in S. pinnata Se had no effect on pollen germination. Floral visitors collected from Se-rich S. pinnata contained up to 270 µg g(-1), concentrations toxic to many herbivores. Indeed, floral visitors showed no visitation preference between high- and low-Se plants. Honey from seleniferous areas contained 0.4-1 µg Se g(-1), concentrations that could provide human health benefits. • This study is the first to shed light on the possible evolutionary cost, through decreased pollen germination in B. juncea, of Se accumulation and has implications for the management of seleniferous areas.

Selective embryo abortion in a perennial tree-legume: a case for maternal advantage of reduced seed number per fruit.

In this study, I analyzed time-course of embryo abortion, positional bias in seed maturation and maternal costs of seed packaging in Cercis canadensis. While basal embryos experience similar rates of abortion as those in other positions during the first week of development, abortion rates peak during the second week. Head start in resource sequestration by stigmatic embryos may explain high rates of basal embryo abortion. Similar seed packaging costs and seed mass for single and multi-seeded pods suggests that maternal parent regulates pod size in accordance with seed numbers per pod investing equally in the surviving offspring. Competition during early developmental period results in the abortion of less competitive embryos allowing for optimal resource investment.

How does breeding system variation modulate sexual antagonism?

The study of sexually antagonistic (SA) traits remains largely limited to dioecious (separate sex), mobile animals. However, the occurrence of sexual conflict is restricted neither by breeding system (the mode of sexual reproduction, e.g. dioecy or hermaphroditism) nor by sessility. Here, we synthesize how variation in breeding system can affect the evolution and expression of intra- and inter-locus sexual conflicts in plants and animals. We predict that, in hermaphrodites, SA traits will (i) display lower levels of polymorphism; (ii) respond more quickly to selection; and (iii) involve unique forms of interlocus conflict over sex allocation, mating roles and selfing rates. Explicit modelling and empirical tests in a broader range of breeding systems are necessary to obtain a general understanding of the evolution of SA traits.

The genetic correlation between flower size and water use efficiency in monkeyflowers.

QUESTION: Does water loss during drought stress represent an important physiological constraint on the evolution of flower size? ORGANISM: A genetically diverse population of Mimulus guttatus (yellow monkeyflower) originally sampled from an alpine meadow in Oregon, USA. METHODS: We grew plants of three different genotypic classes (small, medium, and large flowered) under both well-watered and drought-stress conditions and measured water use efficiency using stable carbon isotopes. RESULTS: There was no difference in water use efficiency among flower size genotypes under well-watered conditions, but the water use efficiency of small-flowered plants was substantially lower than that of medium or large genotypes under drought stress. Whether this paradoxical result is a direct effect of flower size or an indirect (i.e. pleiotropic) effect, the presence of a genetic correlation between floral and physiological traits indicates that selection of one does impact the other.

Bee genera, diversity and abundance in genetically modified canola fields.

Intensive agricultural practices resulting in large scale habitat loss ranks as the top contributing factors in the global bee decline. Growing Genetically Modified Herbicide Tolerant (GMHT) crops as large monocultures has resulted extensive applications of herbicides leading to the degradation of natural habitats surrounding farmlands. Herbicide tolerance trait is beneficial for crops such as Canola (Brassica napus) that are extremely vulnerable to weed competition. While the trait in itself does not harm pollinators, growing genetically modified herbicide tolerant cultivars indirectly contributes towards pollinator declines through habitat loss. Canola, a mass-flowering crop is highly attractive to bee pollinators and the extensive adoption of the herbicide tolerant trait has led to depletion of non-crop floral resources. Extensive use of herbicide in and near fields with herbicide tolerant cultivars systematically eliminates semi-natural habitats around agricultural fields which consist of non-crop flowering plants. Planting pollinator strips provides floral resources for bees after crop flowering. We document the bee genera in canola and the adjoining pollinator strip. The overlap in bee genera reinforces the importance of pollinator habitats in agricultural landscape.

Cooperative nest building and brood care by nestmates and non nestmates in Ropalidia marginata : implications for the evolution of eusociality.

In the primitively eusocial wasp, Ropalidia marginata, low levels of intra-colony genetic relatedness, lack of intra-colony kin discrimination and acceptance of young wasps into alien colonies, prompted us to investigate whether or not there exists a cost of such high genetic variability. Freshly eclosed wasps were paired either with their nestmates or with their non nestmates and their performance in nest building and brood care were compared. There was no demonstrable difference between nestmate and non nestmate pairs in terms of success in raising adult offspring, time required for nest initiation, brood developmental period and productivity. There was also no difference in the efficiency of cooperation and division of labour between the nestmate pairs and non nestmate pairs. These results reinforce the idea that the haplodiploidy hypothesis is insufficient to explain the prevalence of worker behaviour in R. marginata and emphasize the importance of factors other than genetic relatedness in the evolution of eusociality.

Receiver bias for exaggerated signals in honeybees and its implications for the evolution of floral displays.

Mechanistic models of animal signals posit the occurrence of biases on the part of receivers that could be potentially exploited by signallers. Such biases are most obvious when animals are confronted with exaggerated versions of signals they normally encounter. Signalling systems operating in plant-pollinator interactions are among the most highly coevolved, with plants using a variety of floral signals to attract pollinators. A number of observations suggest that pollinators preferentially visit larger floral displays although the benefit of this to either the plant or the pollinator is not always clear. We use a standard dual-choice experimental protocol to show that honeybees display a receiver bias for exaggerated size and colour contrast--two important components of floral signals--even when such signals do not indicate quality. We discuss the implications of this receiver bias for the evolution of floral displays and its possible exploitation by invading alien plants.

Sampling and decision rules used by honey bees in a foraging arena.

Animals must continuously choose among various available options to exploit the most profitable resource. They also need to keep themselves updated about the values of all available options, since their relative values can change quickly due to depletion or exploitation by competitors. While the sampling and decision rules by which foragers profitably exploit a flower patch have attracted a great deal of attention in theory and experiments with bumble bees, similar rules for honey bee foragers, which face similar foraging challenges, are not as well studied. By presenting foragers of the honey bee Apis cerana with choice tests in a foraging arena and recording their behavior, we investigate possible sampling and decision rules that the foragers use to choose one option over another and to track other options. We show that a large part of the sampling and decision-making process of a foraging honey bee can be explained by decomposing the choice behavior into dichotomous decision points and incorporating the cost of sampling. The results suggest that a honey bee forager, by using a few simple rules as part of a Bayesian inference process, is able to effectively deal with the complex task of successfully exploiting foraging patches that consist of dynamic and multiple options.

Inbreeding and the genetic variance in floral traits of Mimulus guttatus.

The additive genetic variance, V(A), is frequently used as a measure of evolutionary potential in natural plant populations. Many plants inbreed to some extent; a notable observation given that random mating is essential to the model that predicts evolutionary change from V(A). With inbreeding, V(A) is not the only relevant component of genetic variation. Several nonadditive components emerge from the combined effects of inbreeding and genetic dominance. An important empirical question is whether these components are quantitatively significant. We use maximum likelihood estimation to extract estimates for V(A) and the nonadditive 'inbreeding components' from an experimental study of the wildflower Mimulus guttatus. The inbreeding components contribute significantly to four of five floral traits, including several measures of flower size and stigma-anther separation. These results indicate that inbreeding will substantially alter the evolutionary response to natural selection on floral characters.

Seed abortion in Pongamia pinnata (Fabaceae).

In Pongamia pinnata only one of the two ovules develops into a seed in most of the pods. Since pollen was not found to be limiting and reduced fertilization could not completely explain the observed frequency of seed abortion, it implied an effect of postfertilization factors. Aqueous extracts of developing seeds and maternal tissue (placenta) did not influence abortion in vitro, suggesting that abortion may not be mediated by a chemical. Experimental uptake of (14)C sucrose in vitro indicated that both the stigmatic and the peduncular seed have similar inherent capacities of drawing resources, but the peduncular seed is deprived of resources in the presence of the stigmatic seed. This deprivation of the peduncular seed could be offset by supplying an excess of hormones leading to the subsequent formation of two seeds in a pod. The prevalence of single-seeded pods in P. pinnata seems therefore to be a result of competition between the two seeds for maternal resources. The evolutionary significance of single-seeded pods in P. pinnata is discussed with respect to possible dispersal advantage enjoyed by such pods.