Pollen Samples Research Articles

Impacts of a tailing dam break on the contamination of pollen and honey from the stingless bee Tetragonisca angustula (Apidae: Trigonini)

Metal and metalloid pollution are one of the main negative impacts of mining activities. Furthermore, the mining wastes are often stored in tailing dams, which are prone to failures and even total collapse. The Fundão tailing dam collapse in Brazil in 2015 flooded large areas of river floodplains below the dam. We analyzed the effect of the tailing deposition from the dam disaster on metal/metalloid levels in honey and pollen samples from the stingless bee Tetragonisca angustula. Pollen and honey samples were taken from the initial stretch of the impacted areas, as well as adjacent control areas and an additional external control area. In total, we analyzed 55 pollen samples and 53 honey samples, which were collected directly from the honey and pollen combs stored within the hives. The samples were wet acid digested in a microwave oven and the concentrations of the elements were determined by ICP-OES and ICP-MS. The metal/metalloid contents in honey and pollen samples were compared between the impacted and the non-impacted points. We found significantly higher concentrations of As, Ba, Cr, Mn, Ni and Zn, in honey and pollen samples, from some impacted points compared to the reference non-impacted ones. However, most concentrations we found were similar to those found in other works, including areas with mining activity in the Iron Quadrangle (Minas Gerais, Brazil). These results suggest that the tailing dram break, followed by deposition of the dam waste, contributed to a high concentration of some elements found in the pollen and honey samples at our study site.

Clopyralid applied to winter oilseed rape (Brassica napus L.) contaminates the food products nectar, honey and pollen

Clopyralid is a systemic herbicide used in oilseed rape and other crops. It was found in Danish honey from 2016 in concentrations exceeding the maximum residue level (MRL) of 0.05 mg kg−1. About 50% of the Danish honey is based on nectar from winter oilseed rape. In 2019 and 2020, winter oilseed rape fields were sprayed with clopyralid just before the assigned spraying deadline. At flowering, nectar and pollen samples were collected and the content of clopyralid was measured. Honey and pollen samples were also collected from beehives next to ten conventional winter oilseed rape fields sprayed with clopyralid. Clopyralid was found in nectar and pollen from the experimental fields, and in honey and pollen from beehives next to the conventional fields. For most samples the content in nectar and honey exceeded the MRL. The concentrations found, may not pose any health risk for consumers, as the MRL is based on the original detection limit and not on toxicological tests. However, it can have a significant economical consequence for the beekeepers, who are not allowed to sell the honey if the concentration of clopyralid exceeds 0.1 mg kg−1. Reducing the acceptable applicable rate of clopyralid or implementing an earlier deadline for spraying of clopyralid may reduce the risk of contaminating bee food products. However, if it is not possible to obtain a satisfactory effect of clopyralid on the weed flora under these conditions, spraying with pesticides containing clopyralid should be restricted in winter oilseed rape. Determination of an MRL value based on toxicological tests might result in a higher value and make it acceptable selling the honey containing higher levels of clopyralid.

Chemical constituents, antioxidant, and anticancer activities of bee pollen from various floral sources in Taiwan

Bee pollen has been traditionally used for health promotion. However, the chemical constituents and pharmaceutical effects of bee pollen strongly depend on their sources. This study determined chemical compositions and evaluated the antioxidant and anticancer activity of six bee pollen samples from Taiwan. The Cs sample contained the highest amounts of carotenoid (417.67 mg/g DW) and anthocyanin (10.96 μmol/g DW) while the Nn sample showed the highest content of chlorophyll (Chl) a (23.39 mg/g DW) and Chl b (39.17 mg/g DW). The highest flavonoid (11.69 mg QE/g DW) and phenolic content (42.91 mg GAE/g DW) were found in Bp and Pm samples, respectively. The highest Fe2+-chelating ability was observed in Bp (IC50 value of 6.28 mg/mL), while Bn exhibited the most effective in scavenging DPPH radical with IC50 value of 3.96 mg/mL. The Bp sample also showed the highest activity against three breast cancer cell lines, MCF-7 (cell viability of 43.5%), BT-20 (cell viability of 0%), and Hs 578T (cell viability of 0%). This study suggested that the level of bioactive compounds and biological activity of bee pollen significantly differ among their sources and the Bp is a potent antioxidant and anticancer agent for medicinal use.

PALYNOLOGICAL ANALYSIS, PHENOLIC COMPONENTS AND ANTI-INFLAMMATORY ACTIVITY OF SOME BEE POLLENS COLLECTED FROM THE NORTHEAST REGION OF ALGERIA

Bee pollen is multiplex blend of floral pollen and nectar agglutinated by bee salivary substances. It is famously known for being high in proteins, carbs, lipids, vitamins, and phenolic compounds, among other physiologically dynamic components. Its composition fluctuates incredibly agreeing to both botanical origins and edaphoclimatic conditions. In this work, the botanical origin, the phenolic components and the anti-inflammatory activity in vivo of eight bee pollens intended for human consumption were taken from distinctive apiaries in Algeria’s northeast, were determined and compared. All samples were detected heterofloral based on the identification of forty pollen types belonging to 22 botanical families. Total phenolic contents varied between 752.94 ± 17.78 and 12247.06 ± 40.04 mg GAE/ 100g, while the total flavonoid contents ranged from 2680.55 ± 12.02 to 8506.94 ± 15.56 mg QE/ 100g, and the total flavonol contents were in the interval between 4978.87 ± 33.39 and 7903.75 ± 24.39 mg QE/ 100g. The obtained results showed that the bulk of the ethanolic extracts had a good anti-inflammatory activity. As a conclusion, all the aforementioned heterofloral bee pollen samples could significantly be a wealthy source of polyphenols with a potential anti-inflammatory activity.

Glucosinolates as Markers of the Origin and Harvesting Period for Discrimination of Bee Pollen by UPLC-MS/MS.

Bee pollen is currently one of the most commonly consumed food supplements, as it is considered to be a good source of bioactive substances and energy. It contains various health-promoting compounds, such as proteins, amino acids, lipids, as well as glucosinolates. In the present study, the glucosinolate content was determined, by means of ultra-performance liquid chromatography coupled to a quadrupole time-of-flight mass detector, in 72 bee pollen samples from four different apiaries in Guadalajara (Spain), harvested in three different periods. In addition, 11 commercial multifloral samples from different Spanish regions were also analyzed. The aim was to verify the suitability of these compounds as biomarkers of their geographical origin, and to test their potential for distinguishing the harvesting period. By means of a canonical discriminant analysis, it was possible to differentiate the apiary of origin of most of the samples, and these could also be clearly differentiated from the commercial ones, simply as a result of the glucosinolate content. In addition, it was also demonstrated for the first time that bee pollen samples were capable of being differentiated according to the time of harvesting and their glucosinolate content.

Bee Pollen Extracts: Chemical Composition, Antioxidant Properties, and Effect on the Growth of Selected Probiotic and Pathogenic Bacteria.

This paper evaluated the chemical and biological properties of bee pollen samples from Romania. Firstly, the bee pollen alcoholic extracts (BPEs) were obtained from raw bee pollen harvested by Apis mellifera carpatica bees. The chemical composition of BPE was obtained by determination of total phenol content and total flavonoid content, UHPLC-DAD-ESI/MS analysis of phenolic compounds, and GC-MS analysis of fatty acids, esters, and terpenes. Additionally, the antioxidant activity was evaluated by the Trolox Equivalent Antioxidant Capacity method. Furthermore, the biological properties of BPE were evaluated (antimicrobial and cytotoxic activity). The raw BP samples studied in this paper had significant phenolic acid and flavonoid content, and moderate fatty acid, ester, and terpene content. P1, P2, and P4 have the highest TPC and TFC levels, and the best antioxidant activity. All BPEs studied had antimicrobial activity on pathogenic strains isolated from the clinic or standard strains. A synergistic antimicrobial effect of the BPEs was observed along with the soluble compounds of L. rhamnosus MF9 and E. faecalis 2M17 against some pathogenic (clinical) strains and, considering the tumour proliferation inhibitory activity, makes BP a potential prebiotic and antitumour agent for the gut environment.

Concentration levels and an assessment of human health risk of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) in honey and pollen.

Persistent organic pollutants (POPs) such as polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) accumulate in the food chain due to their physical and chemical properties and adversely affect human health. For this reason, this study aimed to determine the PAH and PCB concentration levels in pollen and honey samples in urban and semi-urban areas and to evaluate the risk of cancer that may occur by ingestion in Bursa, Turkey. The average total concentrations of 14 PAH (∑14PAH) compounds in pollen and honey samples were found to be 304.3 ± 192.3ng/g (average ± standard deviation) and 650.2 ± 118.1ng/g for the urban area, and 329.6 ± 160.6ng/g and 464.3 ± 66.4ng/g for the semi-urban area, respectively. Similarly, ∑14PCB concentrations in pollen and honey samples were found to be 8.7 ± 3.6ng/g and 13.0 ± 4.8ng/g for the urban area and 7.7 ± 2.2ng/g and 17.4 ± 4.0ng/g for the semi-urban area, respectively. It was determined that the pollen and honey samples in both sampling areas were affected by local PCB sources. The Pearson correlation coefficient (PCC) method determined the relationship between pollen and honey samples. According to the PCC values obtained, it was observed that pollen and honey in both sampling regions exhibited a significant relationship with each other. Finally, while there was no cancer risk for PCBs due to ingestion of honey and pollen in both sampling areas, acceptable cancer risk has been calculated for PAHs.

Pesticide risk to managed bees during blueberry pollination is primarily driven by off-farm exposures

When managed bee colonies are brought to farms for crop pollination, they can be exposed to pesticide residues. Quantifying the risk posed by these exposures can indicate which pesticides are of the greatest concern and helps focus efforts to reduce the most harmful exposures. To estimate the risk from pesticides to bees while they are pollinating blueberry fields, we sampled blueberry flowers, foraging bees, pollen collected by returning honey bee and bumble bee foragers at colonies, and wax from honey bee hives in blooming blueberry farms in southwest Michigan. We screened the samples for 261 active ingredients using a modified QuEChERS method. The most abundant pesticides were those applied by blueberry growers during blueberry bloom (e.g., fenbuconazole and methoxyfenozide). However, we also detected highly toxic pesticides not used in this crop during bloom (or other times of the season) including the insecticides chlorpyrifos, clothianidin, avermectin, thiamethoxam, and imidacloprid. Using LD50 values for contact and oral exposure to honey bees and bumble bees, we calculated the Risk Quotient (RQ) for each individual pesticide and the average sample RQ for each farm. RQ values were considered in relation to the U.S. Environmental Protection Agency acute contact level of concern (LOC, 0.4), the European Food Safety Authority (EFSA) acute contact LOC (0.2) and the EFSA chronic oral LOC (0.03). Pollen samples were most likely to exceed LOC values, with the percent of samples above EFSA’s chronic oral LOC being 0% for flowers, 3.4% for whole honey bees, 0% for whole bumble bees, 72.4% for honey bee pollen in 2018, 45.4% of honey bee pollen in 2019, 46.7% of bumble bee pollen in 2019, and 3.5% of honey bee wax samples. Average pollen sample RQ values were above the EFSA chronic LOC in 92.9% of farms in 2018 and 42.9% of farms in 2019 for honey bee collected pollen, and 46.7% of farms for bumble bee collected pollen in 2019. Landscape analyses indicated that sample RQ was positively correlated with the abundance of apple and cherry orchards located within the flight range of the bees, though this varied between bee species and landscape scale. There was no correlation with abundance of blueberry production. Our results highlight the need to mitigate pesticide risk to bees across agricultural landscapes, in addition to focusing on the impact of applications on the farms where they are applied.

Microbiological characterization of fresh bee pollens from the Aegean region of Turkey

ABSTRACT. The aim of this study is to characterize the microbiological safety of Turkish fresh bee pollen for the first time. Twenty-one fresh bee pollen samples were purchased from local beekeepers between September 2020 and October 2020 in Muğla, İzmir, Kütahya and Afyon provinces in the Aegean region of Turkey. The samples were analyzed for total aerobic bacteria, total coliform bacteria, total psychotropic microorganism, total lactic acid bacteria, Staphyloccus aureus, sulfite-reducing clostridia spores, yeast and mold. Total ochratoxin and aflatoxin levels were measured using lateral flow kit in samples contaminated with mycotoxigenic molds. The counts of the microorganisms ranged from 3.70 to 5.42 log CFU g-1 for the aerobic mesophilic, 3.0 to 5.40 log CFU g -1 for psychrotrophs, 3.85 to 5.78 log CFU g -1 for lactic acid bacteria (LAB), 3.0 to 5.45 log CFU g-1 for yeasts-molds, and <10 to 5.0 log CFU g-1 for total coliforms in the samples. Besides, S. aureus and sulfite reducing clostridia were not detected among the samples. The predominant mold genera in samples were Aspergillus sp. Penicillium sp. and Alternaria sp. Five samples contaminated with mycotoxicogenic molds had aflatoxins in the range of 2.96-9.71 µg/kg. According to the study results, comprehensive prevention, control and surveillance strategies need to be established to reduce bacteria and mycotoxin contaminations in fresh bee pollen. Legal regulation of bee pollen as food supplement and legal limit of mycotoxins in bee pollen should be defined.

Biomonitoring via DNA metabarcoding and light microscopy of bee pollen in rainforest transformation landscapes of Sumatra

BackgroundIntense conversion of tropical forests into agricultural systems contributes to habitat loss and the decline of ecosystem functions. Plant-pollinator interactions buffer the process of forest fragmentation, ensuring gene flow across isolated patches of forests by pollen transfer. In this study, we identified the composition of pollen grains stored in pot-pollen of stingless bees, Tetragonula laeviceps, via dual-locus DNA metabarcoding (ITS2 and rbcL) and light microscopy, and compared the taxonomic coverage of pollen sampled in distinct land-use systems categorized in four levels of management intensity (forest, shrub, rubber, and oil palm) for landscape characterization.ResultsPlant composition differed significantly between DNA metabarcoding and light microscopy. The overlap in the plant families identified via light microscopy and DNA metabarcoding techniques was low and ranged from 22.6 to 27.8%. Taxonomic assignments showed a dominance of pollen from bee-pollinated plants, including oil-bearing crops such as the introduced species Elaeis guineensis (Arecaceae) as one of the predominant taxa in the pollen samples across all four land-use types. Native plant families Moraceae, Euphorbiaceae, and Cannabaceae appeared in high proportion in the analyzed pollen material. One-way ANOVA (p > 0.05), PERMANOVA (R² values range from 0.14003 to 0.17684, for all tests p-value > 0.5), and NMDS (stress values ranging from 0.1515 to 0.1859) indicated a lack of differentiation between the species composition and diversity of pollen type in the four distinct land-use types, supporting the influx of pollen from adjacent areas.ConclusionsStingless bees collected pollen from a variety of agricultural crops, weeds, and wild plants. Plant composition detected at the family level from the pollen samples likely reflects the plant composition at the landscape level rather than the plot level. In our study, the plant diversity in pollen from colonies installed in land-use systems with distinct levels of forest transformation was highly homogeneous, reflecting a large influx of pollen transported by stingless bees through distinct land-use types. Dual-locus approach applied in metabarcoding studies and visual pollen identification showed great differences in the detection of the plant community, therefore a combination of both methods is recommended for performing biodiversity assessments via pollen identification.

Human-vegetation dynamics in Holocene south-eastern Norway based on radiocarbon dated charcoal from archaeological excavations

Charcoal from archaeological contexts differs from off-site pollen samples as it is mainly a product of intentional human action. As such, analysis of charcoal from excavations is a valuable addition to studies of past vegetation and the interaction between humans and the environment. In this paper, we use a dataset consisting of 6186 dated tree species samples from 1239 archaeological sites as a proxy to explore parts of the Holocene forest development and human-vegetation dynamics in South-Eastern Norway. From the middle of the Late Neolithic (from c. 2000 BC) throughout the Early Iron Age (to c. AD 550) the region’s agriculture is characterized by fields, pastures and fallow. Based on our data, we argue that these practices, combined with forest management, clearly altered the natural distribution of trees and favoured some species of broadleaved trees. The past distribution of hazel ( Corylus avellana) is an example of human impact on the vegetation. Today, hazel is not even among the 15 most common tree species, while it is one of the most prevalent species in the archaeological record before AD 550. The data indicate that this species was favoured already by the region’s Mesolithic hunter-fisher-gatherers, and that it was among the species that thrived extremely well in the early farming landscape. Secondly, our analysis also indicates that spruce ( Picea abies) first formed large stands in the south-eastern parts of Norway c. 500 BC, centuries earlier than previously assumed. It is argued that this event, and a further westward expansion of spruce, was partly a consequence of a specific historical event – the first millennium BC farming expansion.

Precision glycerine jelly swab for removing pollen from small and fragile insect specimens

Abstract Historical datasets can establish a critical baseline of plant–animal interactions for understanding contemporary interactions in the context of global change. Pollen is often incidentally preserved on animals in natural history collections. Techniques for removing pollen from insects have largely been developed for fresh insect specimens or historical specimens with large amounts of pollen on specialized structures. However, many key pollinating insects do not have these specialized structures and thus, there is a need for a method to extract pollen from these small and fragile insects. Here, we propose a precision glycerine jelly swab tool to allow for the precise removal of pollen from old, small and fragile insect specimens. We use this tool to remove pollen from five families of insects collected in the late 1970s. Additionally, we compare our method with four previously published techniques for removing pollen from pinned contemporary specimens. We show the functionality of the precision glycerine jelly swab for removing small quantities of pollen across insect families. We found that across the five methods, all removed pollen; yet, it was clear that some are better suited for fragile specimens. In particular, the traditional glycerine jelly swab and the precision glycerine jelly swabs both performed well for removing pollen from bee faces. The shaking wash resulted in specimen fracture and residue left behind, the ethanol rinses left setae matted, and the glycerol swabbing left residue on the specimen. Additionally, we present photographs documenting the effects of these methods on pinned honey bee specimens. The precision glycerine jelly swab opens up opportunities to sample pollen from a variety of insects in natural history collections. These pollen samples can be incorporated into downstream analyses for pollen identification either via microscopy or DNA sequencing, and the resulting plant–insect interaction data can establish historical baselines for contemporary comparison. Beyond our application of this method to pollen on insects, this precision glycerine jelly swab tool could be used to explore pollen placement specialization or to sample bryophyte, fungal and tree fern spores dispersing on animals.

Pollen Viability of Fraxinus excelsior in Storage Experiments and Investigations on the Potential Effect of Long-Range Transport

Fragmented ash populations due to ash dieback may lead to a limited gene flow and pollination success. Therefore, the viability of ash pollen plays a major role for the survival of the species. The extent to which the long-distance transport of pollen affects pollen viability was investigated with experiments in a climate chamber using ash pollen samples from a seed orchard in Emmendingen, Germany. Furthermore, experiments with a volumetric pollen trap were conducted. A suitable storage temperature for ash pollen was determined by using four viability tests; TTC test, pollen germination, Alexander’s stain and Acetocarmine. An optimization of the germination medium was performed. We found a strong influence of prevailing temperatures on pollen viability, which decreased faster under warmer conditions. At moderate temperatures, viable pollen could still be observed after 28 days. Thus, a possible successful pollination can also be associated to long-range transported pollen. Storage experiments showed that pollen viability could be maintained longer at temperatures of −20 °C and −80 °C than at 4 °C. In particular, the TTC test has proven to be suitable for determining viability. Therefore, properly stored pollen can be used for breeding programs to support the survival of Fraxinus excelsior.

European pollen-based REVEALS land-cover reconstructions for the Holocene: methodology, mapping and potentials

Abstract. Quantitative reconstructions of past land cover are necessary to determine the processes involved in climate–human–land-cover interactions. We present the first temporally continuous and most spatially extensive pollen-based land-cover reconstruction for Europe over the Holocene (last 11 700 cal yr BP). We describe how vegetation cover has been quantified from pollen records at a 1∘ × 1∘ spatial scale using the “Regional Estimates of VEgetation Abundance from Large Sites” (REVEALS) model. REVEALS calculates estimates of past regional vegetation cover in proportions or percentages. REVEALS has been applied to 1128 pollen records across Europe and part of the eastern Mediterranean–Black Sea–Caspian corridor (30–75∘ N, 25∘ W–50∘ E) to reconstruct the percentage cover of 31 plant taxa assigned to 12 plant functional types (PFTs) and 3 land-cover types (LCTs). A new synthesis of relative pollen productivities (RPPs) for European plant taxa was performed for this reconstruction. It includes multiple RPP values (≥2 values) for 39 taxa and single values for 15 taxa (total of 54 taxa). To illustrate this, we present distribution maps for five taxa (Calluna vulgaris, Cerealia type (t)., Picea abies, deciduous Quercus t. and evergreen Quercus t.) and three land-cover types (open land, OL; evergreen trees, ETs; and summer-green trees, STs) for eight selected time windows. The reliability of the REVEALS reconstructions and issues related to the interpretation of the results in terms of landscape openness and human-induced vegetation change are discussed. This is followed by a review of the current use of this reconstruction and its future potential utility and development. REVEALS data quality are primarily determined by pollen count data (pollen count and sample, pollen identification, and chronology) and site type and number (lake or bog, large or small, one site vs. multiple sites) used for REVEALS analysis (for each grid cell). A large number of sites with high-quality pollen count data will produce more reliable land-cover estimates with lower standard errors compared to a low number of sites with lower-quality pollen count data. The REVEALS data presented here can be downloaded from https://doi.org/10.1594/PANGAEA.937075 (Fyfe et al., 2022).

Determination of Free Amino Acids in Bee Pollen by Liquid Chromatography with Fluorescence Detection

Bee pollen is one of the hive products that is of most interest today due to its multiple beneficial health properties, making it an increasingly popular food supplement. Bee pollen contains many bioactive compounds, such as fatty acids, vitamins, minerals, proteins, and amino acids, among others. In the present study, the free amino acid content was determined in bee pollen by using liquid chromatography coupled to a fluorescence detector. Sample treatment consisted of a solvent extraction of the free amino acids with ultrapure water and a further centrifugation of the extract, which was repeated twice. After that, it was necessary to perform a pre-column derivatization of the amino acids using a combination of two reagents (o-phthalaldehyde and 9-fluorenylmethyl chloroformate) prior to their separation in a Gemini® C18 reverse phase column in gradient elution mode. The analytical performance was evaluated, and several commercial bee pollen samples were analyzed. Significant differences in the free amino acid profile and concentration, which ranged between 19 and 192 mg/g, were observed depending on the botanical origin of the samples.

Sweeping-micellar electrokinetic chromatography with tandem mass spectrometry as an alternative methodology to determine neonicotinoid and boscalid residues in pollen and honeybee samples

In this work, it is proposed for the first time an electrophoretic approach based on micellar electrokinetic chromatography coupled with tandem mass spectrometry (MEKC-MS/MS) for the simultaneous determination of nine neonicotinoids (NNIs) together with the fungicide boscalid in pollen and honeybee samples. The separation was performed using ammonium perfluorooctanoate (50 mM, pH 9) as both volatile surfactant and electrophoretic buffer compatible with MS detection. A stacking strategy for accomplishing the on-line pre-concentration of the target compounds, known as sweeping, was carried out in order to improve separation efficiency and sensitivity. Furthermore, a scaled-down QuEChERS was developed as sample treatment, involving a lower organic solvent consumption and using Z-Sep+ as dispersive sorbent in the clean-up step. Regarding the detection mode, a triple quadrupole mass spectrometer was operating in positive ion electrospray mode (ESI+) under multiple reaction monitoring (MRM). The main parameters affecting MS/MS detection as well as the composition of the sheath-liquid (ethanol/ultrapure water/formic acid, 50:49.5:0.5 v/v/v) and other electrospray variables were optimized in order to achieve satisfactory sensitivity and repeatability. Procedural calibration curves were established in pollen and honeybee samples with LOQs below 11.6 µg kg−1 and 12.5 µg kg−1, respectively. Precision, expressed as RSD, lower than 15.2% and recoveries higher than 70% were obtained in both samples. Two positive samples of pollen were found, containing imidacloprid and thiamethoxam. Imidacloprid was also found in a sample of honeybees. The obtained results highlight the applicability of the proposed method, being an environmentally friendly, efficient, sensitive and useful alternative for the determination of NNIs and boscalid in pollen and honeybee samples.

SEC1A is a major Arabidopsis Sec1/Munc18 gene in vesicle trafficking during pollen tube tip growth.

SUMMARYPollen tubes (PTs) grow by the targeted secretion of new cell wall material to their expanding tip region. Sec1/Munc18 (SM) proteins promote membrane fusion through regulation of the SNARE complex. We have previously shown that disruption of protein glycosylation in the Arabidopsis thaliana hpat1 hpat3 double mutant leads to PT growth defects that can be suppressed by reducing secretion. Here, we identified five point mutant alleles of the SM protein SEC1A as hpat1/3 suppressors. The suppressors increased seed set, reduced PT growth defects and reduced the rate of glycoprotein secretion. In the absence of the hpat mutations, sec1a reduced pollen germination and PT elongation producing shorter and wider PTs. Consistent with a defect in membrane fusion, sec1a PTs accumulated secretory vesicles. Though sec1a had significantly reduced male transmission, homozygous sec1a plants maintained full seed set, demonstrating that SEC1A was ultimately dispensable for pollen fertility. However, when combined with a mutation in another SEC1‐like SM gene, keule, pollen fertility was totally abolished. Mutation in sec1b, the final member of the Arabidopsis SEC1 clade, did not enhance the sec1a phenotype. Thus, SEC1A is the major SM protein promoting pollen germination and tube elongation, but in its absence KEULE can partially supply this activity. When we examined the expression of the SM protein family in other species for which pollen expression data were available, we found that at least one Sec1‐like protein was highly expressed in pollen samples, suggesting a conserved role in pollen fertility in other species.

Evaluation of Environmental and Nutritional Aspects of Bee Pollen Samples Collected from East Black Sea Region, Turkey, via Elemental Analysis by ICP-MS.

Honeybee pollens are good food sources in terms of their mineral contents and are specific to the regions they are collected. In addition, they may be used as bioindicators in the assessment of environmental pollution based on their potentially toxic element contents. In the present study, mineral element composition and potentially toxic element levels of honeybee pollen samples collected from various cities in East Black Sea Region of Turkey (18 samples) were determined by inductively coupled plasma mass spectrometry (ICP-MS) after microwave assisted acid digestion. The method validation was performed by using CRM (Certified Reference Material-BCR®279-Sea Lettuce-Ulva lactuca) to evaluate the accuracy and precision. Elemental composition of honeybee pollens were detected within the following ranges (minimum-maximum, mg kg-1 dry pollen); Mn (manganese): 11.579-117.349, Fe (Iron): 34.865-811.043, Zn (zinc): 17.707-56.223, Se (selenium): 0.422-0.722, Cr (chromium): 0.848-6.949, Cu (copper): 7.510-26.344, Mg (magnesium): 549.921-2149.716, Ca (calcium): 726.575-2201.837, Na (sodium): 36.518-120.283, Pb (lead): < 0.005-0.622, Cd (cadmium): 0.039-1.390, Ni (nickel): 2.317-21.710, and As (arsenic): 1.331-2.248. Recommended daily allowance, target hazard quotients, hazard index, and carcinogenic risk values of the pollens were calculated with the help of these results. In considering THQ values, pollens were determined to be safe for the consumption of both genders. Based on the carcinogenic risk calculation, most of the pollens examined in this study were categorized as moderately risky. Monitoring studies can be used to identify new sources of contamination or the origin and spread of a particular element. Hence, bee pollens can also be considered as potential bioindicators of toxic metal pollution. HIGHLIGHTS: • Mineral content and potentially toxic metal levels of 18 honeybee pollens were determined. • Recommended daily allowance (RDA) values were calculated. • The nutritional aspects of honeybee pollen samples were evaluated. • Hazard quotient (HQ), hazard index (HI), and carcinogenic risk (CR) estimation of honeybee pollens were assessed. • The potentiality of honeybee pollens as a bioindicator for pollution was discussed.

Seasonality and Landscape Composition Drive the Diversity of Pollen Collected by Managed Honey Bees

The western honey bee, Apis mellifera, is the most important and widespread managed pollinator species. Honey bee diet is based on nectar and pollen, and pollen diversity and composition, in particular, affect colony health and fitness. As landscape composition is strongly linked to floral resource heterogeneity, it could influence the resource intake of honey bees. This work aimed to explore how the composition of pollen collected by honey bees was modulated by seasonality and landscape composition heterogeneity in a mountainous cultivated area of Northern Italy. We selected 13 locations, and at each location, we placed two honey bee colonies from which we collected pollen samples every month during the whole flowering season for two consecutive years. We then analyzed pollen samples in the laboratory and determined the Shannon diversity index of each pollen sample and the temporal pollen taxon replacement. We extracted the cover of the main habitat types at three spatial scales and tested the effect of landscape diversity and composition using Principal Component Analysis. Honey bees foraged on a high number of floral resources, however, they mostly collected pollen from a small number of taxa, with pollen type composition changing throughout the flowering season. In early spring and late summer, most pollen grains were collected from a few plant species, while from May to August the number of collected pollen types was significantly higher. Landscape composition affected pollen diversity only at the end of the flowering season. While honey bees were able to collect highly diverse pollen throughout spring and summer regardless of landscape composition, in late summer, when pollen collected is fundamental for the overwintering of the colony and its development in the following season, semi-natural areas became crucial for honey bee foraging activities, with pollen diversity increasing with increasing percentages of semi-natural areas. Our research highlighted the importance for honey bees of certain seasonal resources and of semi-natural habitats at the end of the flowering season, which ensure the subsistence of their colonies throughout the year.

Pollen-insect interaction meta-networks identify key relationships for conservation in mosaic agricultural landscapes.

Flower visitors use different parts of the landscape through the plants they visit, however these connections vary within and among land uses. Identifying which flower‐visiting insects are carrying pollen, and from where in the landscape, can elucidate key pollen–insect interactions and identify the most important sites for maintaining community‐level interactions across land uses. We developed a bipartite meta‐network, linking pollen–insect interactions with the sites they occur in. We used this to identify which land‐use types at the site‐ and landscape‐scale (within 500 m of a site) are most important for conserving pollen–insect interactions. We compared pollen–insect interactions across four different land uses (remnant native forest, avocado orchard, dairy farm, rotational potato crop) within a mosaic agricultural landscape. We sampled insects using flight intercept traps, identified pollen carried on their bodies and quantified distinct pollen–insect interactions that were highly specialized to both natural and modified land uses. We found that sites in crops and dairy farms had higher richness of pollen–insect interactions and higher interaction strength than small forest patches and orchards. Further, many interactions involved pollinator groups such as flies, wasps, and beetles that are often under‐represented in pollen–insect network studies, but were often connector species in our networks. These insect groups require greater attention to enable wholistic pollinator community conservation. Pollen samples were dominated by grass (Poaceae) pollen, indicating anemophilous plant species may provide important food resources for pollinators, particularly in modified land uses. Field‐scale land use (within 100 m of a site) better predicted pollen–insect interaction richness, uniqueness, and strength than landscape‐scale. Thus, management focused at smaller scales may provide more tractable outcomes for conserving or restoring pollen–insect interactions in modified landscapes. For instance, actions aimed at linking high‐richness sites with those containing unique (i.e., rare) interactions by enhancing floral corridors along field boundaries and between different land uses may best aid interaction diversity and connectance. The ability to map interactions across sites using a meta‐network approach is practical and can inform land‐use planning, whereby conservation efforts can be targeted toward areas that host key interactions between plant and pollinator species.