Pollination efficiencies of three bee (Hymenoptera: Apoidea) species visiting rabbiteye blueberry

Inadequate bee pollination limits rabbiteye blueberry, Vaccinium ashei Reade, production in the some areas of the southeastern United States. Honey bees, Apis mellifera L., are currently the only manageable pollinators available for pollinating V. ashei. However, a new adaptable pollinator for rabbiteye blueberry, Osmia ribifloris Cockerell, was successfully reared and flown in captivity. The bee nested successfully in wooden shelters and conferred superior fruit set to 2-yr-old potted, rabbiteye blueberry bushes. Pollination efficiency or the percentage of blueberry flowers to set fruit after being visited once by a female O. ribifloris was comparable to that of the female blueberry bee Habropoda laboriosa (F.) and worker honey bees. Interestingly, honey bees once thought to be inefficient pollinators of rabbiteye blueberry were found to be very efficient, especially for 'Climax' and 'Premier' flowers.     

Coevolved reproductive strategies in the oligolectic bee Callonychium petuniae (Apoidea, Andrenidae) and three purple flowered Petunia species (Solanaceae) in southern Brazil

Females of Callonychium petuniae were found to be oligolectic foragers on three sympatric purple flowered Petunia species in Rio Grande do Sul, southern Brazil. Males patrol patches of Petunia flowers in search for females. They also wait for females in or on the flowers and perch for mates in the vicinity of blossoms. Copulation is always initiated on Petunia flowers. Flying in copulation posture, the couple visits Rrther flowers where the female continues to collect pollen. The mates land outside the Petunia patch to copulate for up to 20 min. and then take ofr separately. We interprete the flights in copula posture as male strateies to prevent the female from returning to the area of high competitor abundance for further pollen collecting after copulation. Thereby males may increase their chances to be the last male to mate before the eggs are fertilized. Femals and males of C. petuniae are the most frequent pollinators of purple flowered Petunia species and indiscriminately transfer pollen between them. Cytogenetic analyses of chromosome numbers and meiotic phases in individuals of two sympatric Petunia species and in presumable hybrids showed, however, that the species maintain their integrity. Spontaneous color choices in multicolored patches of dummy flowers revealed that C. petuniae bees have a high preference for Petunia-purple color tones, as seen by the human eye. We discuss the significance of color selectivity by C. petuniae bees and of the visual signals exhibited by purple flowered Petunia species to keep the coevolved reproductive strategies stable.     

Diversity and effect of historical inundation on bee and wasp (Hymenoptera: Apoidea,Vespoidea) communities in the Brazilian Pantanal

Hymenoptera are the third largest insect order and is one of the most important ecological agents in terrestrial ecosystems, while terrestrial flooded areas are considered priority for conservation due to their uniqueness. We present the first inventory for bees (Apoidea) and wasps (Vespoidea) and describe their alpha, beta, and gamma diversity s in the Brazilian Pantanal to evaluate the effect of the flood intensity on the community structure. We tested the hypothesis that areas with different profiles would present less similar composition, for this, sampling was carried out in the Brazilian Pantanal between November 2015 and March 2016 using passive traps and active search with entomological net in 19 areas distributed in five flood categories. Ecological metrics were used to describe the community, as well as multivariate analyzes for community interpretation. We collected 3342 individuals belonging to 377 species and eight families and expected 532 (±?23) species of Apoidea and Vespoidea. Different areas presented low similarity and the compositions of the communities tended to be different, mainly between the extremes of the flood profile for different families. The biological characteristics of representatives from the highest taxonomic levels (Subfamily, Tribe) are important for occupation of the areas in the Pantanal. Describing the fauna and understanding how it is influenced by flooding history provides tools for conservation strategies in the Pantanal and further effort is needed to preserve distinct areas within the biome.     

Modelling and estimating pollen movement in oilseed rape (Brassica napus) at the landscape scale using genetic markers

Understanding patterns of pollen movement at the landscape scale is important for establishing management rules following the release of genetically modified (GM) crops. We use here a mating model adapted to cultivated species to estimate dispersal kernels from the genotypes of the progenies of male-sterile plants positioned at different sampling sites within a 10 x 10-km oilseed rape production area. Half of the pollen clouds sampled by the male-sterile plants originated from uncharacterized pollen sources that could consist of both large volunteer and feral populations, and fields within and outside the study area. The geometric dispersal kernel was the most appropriate to predict pollen movement in the study area. It predicted a much larger proportion of long-distance pollination than previously fitted dispersal kernels. This best-fitting mating model underestimated the level of differentiation among pollen clouds but could predict its spatial structure. The estimation method was validated on simulated genotypic data, and proved to provide good estimates of both the shape of the dispersal kernel and the rate and composition of pollen issued from uncharacterized pollen sources. The best dispersal kernel fitted here, the geometric kernel, should now be integrated into models that aim at predicting gene flow at the landscape level, in particular between GM and non-GM crops.     

Diversity in bee (Hymenoptera: Apoidea) and social wasp (Hymenoptera: Vespidae, Polistinae) community in "campos rupestres", Bahia, Brazil

Hymenoptera such as bees and social wasps are regular floral visitors in "campos rupestres" vegetation. A community of bees and social wasps was studied during floral visitation in an area of "campos rupestres", at Chapada Diamantina, BA, Brazil, from September 2001 to April 2002. The community was described in relation to diversity, evenness, and dominance rank, considering the individuals abundance (H' = 2.14/ J' = 0.55) and biomass (H' = 2.34/ J' = 0.60). Thirty nine bee (588 individuals/ 15.742 g) and 11 social wasp species (52 individuals/ 2.156 g) were collected, being the first report of social wasps for the Brazilian "campos rupestres". The main species regarding number of individuals were Trigona spinipes (Fabricius), Apis mellifera L., Frieseomelitta francoi (Moure), and Bombus brevivillus Franklin. About 48% of the species were represented by a single individual. There was an inversion in the dominance rank when the species biomass was considered. B. brevivillus, A. mellifera, T spinipes, and other species represented by 15 individuals or less, such as the social wasps Synoeca cyanea (Olivier), Polistes canadensis (L.) and Myschocyttarus drewseni (Saussure), and the bees Eufriesea nigrohirta (Friese), Xylocopa grisescens Lepeletier and Megachile (Pseudocentron) sp.l were the predominant species. The use of biomass in diversity analysis permitted to detect differences in the relative contribution of species in hierarchy dominance. The comparison between bee faunas from different areas indicates a large similarity of the sampled fauna in Palmeiras (Bahia State) with neighboring ecosystems, although with low values of similarity.     

Glucosinolates in oilseed rape (Brassica napus) and the incidence of pollen beetles (Meligethes aeneus)

During an early stage of migration into a field of oilseed rape in the spring of 1988, more pollen beetles were observed on the double-low variety, Ariana, than on the adjacent single-low varieties, Bienvenu and Mikado. More beetles were found on open flowers on terminal racemes than on yellow and green flower buds on lateral racemes. Five alkenyl, three indolyl and two aromatic glucosinolates were identified in the floral tissues of oilseed rape. The major compounds were progoitrin (2-OH, 3-butenyl) and glucobrassicanapin (4-pentenyl). There were no differences in the types and concentrations of glucosinolates present in the terminal and lateral racemes of any variety, and only small differences between single- and double-low varieties. Concentrations of glucosinolates in floral tissues of crops grown with a little nitrogen were twice those of well-fertilised crops. There was no association between the number and distribution of pollen beetles on plants and the amounts or types of glucosinolates present in floral tissues.     

Assessing bee (Hymenoptera: Apoidea) diversity of an Illinois restored tallgrass prairie: methodology and conservation considerations

Bee species diversity and the effectiveness of four sampling methods were investigated in a west-central Illinois restored tallgrass prairie. Bees were sampled using malaise traps, ground-level pan traps, elevated pan traps, and vane traps. A total of 4,622 bees representing 31 genera and 111 species were collected. Malaise traps collected the greatest number of bees and species, and ground-level pan traps the least. Among the pan traps and vane traps, blue-colored traps collected the greatest abundance and species richness, and yellow traps the least. Chao1 estimator and rarefaction analyses showed that substantial increases in sample sizes would be necessary to achieve asymptotic species richness levels, particularly if ground-level pan traps alone were used. Elevated pan traps and vane traps collected relatively similar species composition. Different colored pan traps at the same height collected more similar species composition than did those at different heights, but species composition of blue ground-level pan traps was relatively similar to elevated pan traps, regardless of color. Indicator species analysis revealed 22 species that were significantly associated with a specific trap type, and 11 species that were associated with a particular pan trap color/elevation. Results of this study show that elevated traps can increase the effectiveness of bee surveys in tallgrass prairie, and that a combination of trap types gives a more complete picture of the bee fauna than does a single survey method. These results should be considered along with cost, ease of use, and goals when planning and designing bee inventories.     

Potential effects of oilseed rape expressing oryzacystatin-1 (OC-1) and of purified insecticidal proteins on larvae of the solitary bee Osmia bicornis

Despite their importance as pollinators in crops and wild plants, solitary bees have not previously been included in non-target testing of insect-resistant transgenic crop plants. Larvae of many solitary bees feed almost exclusively on pollen and thus could be highly exposed to transgene products expressed in the pollen. The potential effects of pollen from oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) were investigated on larvae of the solitary bee Osmia bicornis (= O. rufa). Furthermore, recombinant OC-1 (rOC-1), the Bt toxin Cry1Ab and the snowdrop lectin Galanthus nivalis agglutinin (GNA) were evaluated for effects on the life history parameters of this important pollinator. Pollen provisions from transgenic OC-1 oilseed rape did not affect overall development. Similarly, high doses of rOC-1 and Cry1Ab as well as a low dose of GNA failed to cause any significant effects. However, a high dose of GNA (0.1%) in the larval diet resulted in significantly increased development time and reduced efficiency in conversion of pollen food into larval body weight. Our results suggest that OC-1 and Cry1Ab expressing transgenic crops would pose a negligible risk for O. bicornis larvae, whereas GNA expressing plants could cause detrimental effects, but only if bees were exposed to high levels of the protein. The described bioassay with bee brood is not only suitable for early tier non-target tests of transgenic plants, but also has broader applicability to other crop protection products.     

Hybridisation between oilseed rape (Brassica napus) and tetraploid Brassica rapa under field conditions

Cross-compatible relatives of crop species contribute to the uncertainty regarding the potential risk of transgene escape from genetically modified varieties. The most successful crossing partner of oilseed rape (Brassica napus L.) is diploid Brassica rapa L. Variation of ploidy level among B. rapa cultivars has, until recently, been neglected in the context of gene flow and hybridisation with oilseed rape. We estimated the extent of hybridisation between autotetraploid B. rapa varieties (female) and B. napus (pollen donor) under experimental field conditions. Morphology, variation of relative DNA amount, and microsatellite markers were used to distinguish between intraspecific offspring of tetraploid B. rapa and interspecific hybrids with B. napus. Of 517 seed progenies of tetraploid B. rapa, 45 juvenile plants showed species specific morphological traits of oilseed rape. The detection of putative hybrids based on variation in relative DNA amounts was problematic due to the occurrence of aneuploidy. In total, 84 offspring showed relative DNA amounts deviating from tetraploid B. rapa, four of which were hexaploids. Of the 205 offspring analysed at three microsatellite loci, 67 had oilseed rape alleles. Based on molecular evidence a minimum hybridisation rate of 13.0% was estimated. A few mother plants accounted for the majority of hybrids. The mean pollen viability of hybrids between B. napus and tetraploid B. rapa (80.6%) was high in comparison with mean pollen viability of triploid hybrids between B. napus and diploid B. rapa. Therefore, the occurrence of tetraploid B. rapa should be taken into consideration when estimating the likelihood of gene flow from oilseed rape to close relatives at the landscape level. Tetraploid B. rapa is a common component of several seed mixtures and establishes feral populations in northwest Germany. Assuming a similar abundance of diploid and tetraploid B. rapa, gene flow from B. napus to tetraploid may be more likely than gene flow to diploid B. rapa.     

Genetic manipulation in cultivars of oilseed rape (Brassica napus) using Agrobacterium

Summary The response of oilseed rape cultivars to infection with Agrobacterium tumefaciens and A. rhizogenes and the possibility of regenerating genetically transformed oilseed rape plants were examined. The frequency at which Agrobacterium induced galls or hairy-roots on in vitro cultured plants ranged from 10% to 70%, depending on the cultivar. From galls induced by the tumorigenic strain T37, known to be strongly shoot inducing on tobacco, roots developed frequently. Occasionally, shoots formed and some of these produced tumour cell specific nopaline. Attempts to grow the transformed shoots into plants have so far been unsuccessful. Whole plants transformed with Ri-T-DNA, however, were regenerated. These had crinkled leaves and abundant, frequently branching roots that showed reduced geotropism, similar to previously isolated Ri T-DNA transformed tobacco and potato plants. The transformed oilseed rape plants flowered, but failed to form seeds.     

Convergent evolution of morphological specializations in Central European bee and honey wasp species as an adaptation to the uptake of pollen from nototribic flowers (Hymenoptera, Apoidea and Masaridae)

In the Central European apoid and masarid fauna, 13 bee species belonging to seven different taxonomic groups and one honey wasp species were found to be equipped with a specialized pollen-collecting apparatus on the face which is used to harvest pollen from nototribic flowers, viz. representatives of the Lamiaceae and the Scrophulariaceae. Pollen from these two plant families plays an important part in the larval nourishment of these hymenopteran species. The pollen-collecting apparatus consists of a peculiar facial pilosity composed of transformed hairs which are thickened at their base and either knobbed apically (Celonites abbreviatus) or extended into a thin tail which is either straight and bent at right angles ( Rophites spp., Anthophora borealis, A. furcata ) or wavy ( Anthidium spp., Anthocopa andrenoides, Osmia aurulenta, O. caerulescens ). Nototribic flowers are worked by pollen-collecting females by rubbing the facial area covered with the transformed hairs over the anthers (Celonites, Anthidium, Anthocopa, Osmia) , by buzzing the pollen amongst the transformed hairs (Anthophora Jurcata) or by a combination of rubbing movements and buzzing (Rophites) . Bee species lacking special morphological devices were observed to perform specialized behaviours when collecting pollen at nototribic flowers. Nototribic flowers, therefore, appear to be difficult to exploit for pollen. The raised position of the anthers in nototribic flowers as a possible means to reduce excessive pollen losses caused by pollen-collecting hymenopterans is discussed.     

Biological control of Sclerotinia disease by Aspergillus sp. on oilseed rape in the field

Sclerotinia sclerotiorum causes serious yield losses to many crops worldwide. Aspergillus sp. Asp-4, previously shown to inhibit germination of sclerotia of S. sclerotiorum in vitro and in the field, was evaluated in field trials for suppression of this pathogen on oilseed rape. Spray application of Asp-4 to the soil prior to sowing rice in a rice–oilseed rape rotation resulted in a significant reduction in incidence of Sclerotinia stem rot on oilseed rape compared with the non-treated control in two field trials. This application of Asp-4 also resulted in a significant reduction in germination of sclerotia relative to the non-treated control in these field trials, suggesting that this reduction in sclerotial germination led to disease control. Microscopic examination demonstrated that Asp-4 could effectively colonise external and internal portions of sclerotia of S. sclerotiorum in vitro. Incubation of Asp-4 with sterile sclerotial material induced production of β-glucanase and chitinase activities by this isolate; β-glucanase and chitinase being potentially capable of degrading the glucan and chitin polymeric components of sclerotia. Incubation of Asp-4 with sterile sclerotial material also resulted in a significant reduction in dry weight of this sclerotial material relative to the non-treated control in 96?h in vitro experiments. Experiments reported here indicate that Aspergillus sp. Asp-4 has promise as a biological control agent for S. sclerotiorum on oilseed rape. Experiments reported here suggest that disease control results from inhibition of germination of sclerotial resting structures due to mycoparasitic colonisation by Asp-4.     

Profiling of phenylpropanoids in transgenic low-sinapine oilseed rape (Brassica napus)

A dsRNAi approach silencing a key enzyme of sinapate ester biosynthesis (UDP-glucose:sinapate glucosyltransferase, encoded by the UGT84A9 gene) in oilseed rape (Brassica napus) seeds was performed to reduce the anti-nutritive properties of the seeds by lowering the content of the major seed component sinapine (sinapoylcholine) and various minor sinapate esters. The transgenic seeds have been produced so far to the T6 generation and revealed a steady suppression of sinapate ester accumulation. HPLC analysis of the wild-type and transgenic seeds revealed, as in the previous generations, marked alterations of the sinapate ester pattern of the transformed seeds. Besides strong reduction of the amount of the known sinapate esters, HPLC analysis revealed unexpectedly the appearance of several minor hitherto unknown rapeseed constituents. These compounds were isolated and identified by mass spectrometric and NMR spectroscopic analyses. Structures of 11 components were elucidated to be 4-O-glucosides of syringate, caffeyl alcohol and its 7,8-dihydro derivative as well as of sinapate and sinapine, along with sinapoylated kaempferol glycosides, a hexoside of a cyclic spermidine alkaloid and a sinapine derivative with an ether-bridge to a C₆-C₃-unit. These results indicate a strong impact of the transgenic approach on the metabolic network of phenylpropanoids in B. napus seeds. Silencing of UGT84A9 gene expression disrupt the metabolic flow through sinapoylglucose and alters the amounts and nature of the phenylpropanoid endproducts.     

Effects of simulated acid rain on oilseed rape (Brassica napus) physiological characteristics at flowering stage and yield

A pot experiment was conducted to study the effects of different acidity simulated acid rain on the physiological characteristics at flowering stage and yield of oilseed rape (B. napus cv. Qinyou 9). Comparing with the control (pH 6.0), weak acidity (pH = 4.0-5.0) simulated acid rain stimulated the rape growth to some extent, but had less effects on the plant biomass, leaf chlorophyll content, photosynthetic characteristics, and yield. With the further increase of acid rain acidity, the plant biomass, leaf chlorophyll content, photosynthetic rate, antioxidative enzyme activities, and non-enzyme antioxidant contents all decreased gradually, while the leaf malonyldialdehyde (MDA) content and relative conductivity increased significantly. As the results, the pod number per plant, seed number per pod, seed weight, and actual yield decreased. However, different yield components showed different sensitivity to simulated acid rain. With the increasing acidity of simulated acid rain, the pod number per plant and the seed number per pod decreased significantly, while the seed weight was less affected.     

模拟酸雨对油菜花期生理特性和产量的影响

以甘蓝型油菜品种秦优九号为材料,通过盆栽试验研究了不同pH值模拟酸雨处理对油菜花期生理特性和产量的影响.结果表明:当模拟酸雨pH=4.0～5.0时,在一定程度上刺激了油菜的生长,植株的生物量、叶绿素含量、光合生理特性和产量构成因素等指标均略高于对照(pH 6.0)或与对照相当,但差异不显著.随着模拟酸雨酸度的增强,植株的生物量、叶绿素含量、光合生理特性、抗氧化酶活性和非酶抗氧化剂含量等生理指标逐渐下降,且随酸度增强下降幅度增大,丙二醛含量和相对电导率显著增加,膜脂过氧化加剧,最终导致单株角果数、每角粒数、粒重和实际产量等指标逐渐下降.但不同产量构成因素对酸雨敏感性不同,单株角果数、每角粒数随酸雨酸度的增强而显著下降,而粒重受酸雨影响不显著.     

Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus)

The objective of this study was to evaluate pollen dispersal inBrassica napus (oilseed rape). The selectable marker, used to follow pollen movement, was a dominant transgene (bar) conferring resistance to the herbicide glufosinate-ammonium. Transgenic and non-transgenic plants of the cultivar Westar were planted in a 1.1 ha field trial, with the transgenic plants in a 9 m diameter circle at the centre, surrounded by non-transgenic plants to a distance of at least 47 m in all directions. A 1 m circle of non-transgenic plants was sown in the centre of the transgenic area to allow estimation of the level of pollen dispersal when plants were in close contact. Honeybee hives were placed at the trial site to optimize the opportunity for cross-pollination. During the flowering period, regular observations were made of the number of plants flowering and the number and type of insects present in 60 1 m² areas. These areas were located uniformly around the plot at distances of 1, 3, 6, 12, 24, 36 and 47 m from the edge of the 9 m circle of transgenic plants. Seed samples were harvested from each of the 7 distances so that approximately 20% of the circumference of the plot was sampled at each distance. The centre non-transgenic circle was also sampled. Plants were grown from the seed samples and sprayed with glufosinate to estimate the frequency of pollen dispersal at each distance. In order to screen enough samples to detect low frequency cross-pollination events, seed samples were tested in the greenhouse and on a larger scale in the field. Results were confirmed by testing progeny for glufosinate resistance and by Southern blot analysis. The estimated percentage of pollen dispersal in the non-transgenic centre circle was 4.8%. The frequency was estimated to be 1.5% at a distance of 1 m and 0.4% at 3 m. The frequency decreased sharply to 0.02% at 12 m and was only 0.00033% at 47 m. No obvious directional effects were detected that could be ascribed to wind or insect activity.     

Electron microscopic structure of the cephalic gland of the bee Andrena variabilis Smith (Hymenoptera: Apoidea)

The structure of the glandular tissue located beneath the eye sulcus in Andrena variabilis Smith was studied by means of light and electron microscopy. The glandular tissue was found to be constituted of single-layered, high cylindrical epithelial cells. Electron microscopy revealed strong foldings of the cell membrane on the basal pole of the gland cells. Next to the basal lamina, nerve fibres were found with great frequency. The central, perinuclear zone of the gland cells was rich in cell organelles. The lamellae of the rough-surfaced endoplasmic reticulum (rer) were mostly observed near the nuclear membrane. The vesicles and tubules of the smooth-surface endoplasmic reticulum (ser) were found to be congregated in contiguous compact regions. A well-developed and complicated canalicular system was enclosed between the rer and ser compartments, presumably serving as a passage for the secretory products. Membrane-bound electron-dense granules (secretory granules) were also a characteristic finding in the ser. In a small proportion of the gland cells, strong vacuolization indicated the degradation processes. From the light and electron microscopic characteristics of the gland cells, the existence of pheromone-producing secretory activity of exocrine type in the cephalic gland is presumed.