Volatile compounds and palynological analysis from pollen pots of stingless bees from the mid-north region of Brazil

ABSTRACT Samburá is the botanical pollen nectar agglutinated by salivary secretions of bees. Stingless bee pollen samples were collected in three periods of the year in Monsenhor Gil town, PI, Brazil, for extraction of volatile constituents by different techniques, analyzed by gas chromatography-mass spectrometry (GC-MS) and the palynological analysis used to identify the dominant pollen. Among the volatile compounds identified, kaur-16-ene, methyl and ethyl hexadecanoate, methyl linoleate and heneicosane were identified more frequently in the studied parameters: period of sample collection and extraction techniques used. The palynological analysis identified the pollen of Mimosa caesalpiniifolia Benth. as the dominant pollen in all samples studied.

Morphological characterization of pollens from three Apiaceae species and their ingestion by twelve-spotted lady beetle (Coleoptera: Coccinellidae) / Caracterização morfológica de três espécies de Apiaceae e sua ingestão pela joaninha de doze pintas (Coleoptera: Coccinellidae)

Abstract Larvae and adults of certain species of predator lady beetles feed on pollen, guaranteeing their survival, and at times, reproduction in the absence of preferred prey. Palynology, therefore, may contribute in the investigation of botanical families visited by these predators in order to obtain this floral resource. There are records of the visitation of Apiaceae flowers by Coleomegilla maculata DeGeer, 1775 (Coleoptera, Coccinellidae), but not the ingestion of their pollen grains by this lady beetle. The external morphology of pollen grains of three Apiaceae aromatic species (Anethum graveolens L., Coriandrum sativum L., Foeniculum vulgare Mill.) was characterized, and it was evaluated the ingestion of these pollens by fourth instar larvae and adults of C. maculata upon confinement along with flowers of these Apiaceae for 24 and 48 hours. The pollen grains of those species presented similar external morphology. In the two times of exposure, the larvae ingested the same amount of pollen from the three Apiaceae species, and the amount of C. sativum pollen ingested was the same between larvae and adults. The amount of A. graveolens pollen grains ingested by the adults was significantly greater than the pollens of C. sativum and F. vulgare, in 24 hours, with the opposite occurring in 48 hours. In the first 24 hours, the adults ingested more A. graveolens pollen than the larvae, with the opposite occurring with F. vulgare. There was no significant difference in the amount of Apiaceae pollen ingested between larvae and adults in 48 hours. The results suggest that the pollen-eating habits of certain aphidophagous lady beetles may be crucial in their preservation within agro-ecosystems.

Resumo Larvas e adultos de certas espécies de joaninhas predadoras podem se alimentar de pólen, garantindo a sua sobrevivência, e, às vezes, a reprodução na ausência da presa preferencial. Palinologia, portanto, pode contribuir na investigação de famílias botânicas visitadas por estes predadores a fim de obter esse recurso floral. Há registros da visitação de flores de Apiaceae por Coleomegilla maculata DeGeer, 1775 (Coleoptera, Coccinellidae), mas não da ingestão de seus grãos de pólen por essa joaninha. A morfologia externa de grãos de pólen de três espécies aromáticas de Apiaceae (Anethum graveolens L., Coriandrum sativum L., Foeniculum vulgare Mill.) foi caracterizada e avaliou-se a ingestão desses polens por larvas de quarto instar e adultos de C. maculata em confinamento com flores dessas Apiaceae por 24 e 48 horas. Os grãos de pólen dessas espécies apresentaram morfologia externa semelhante. Nos dois tempos de exposição, as larvas ingeriram a mesma quantidade de pólen das três espécies de Apiaceae, e a quantidade de pólen de C. sativum ingerida foi semelhante entre as larvas e adultos. A quantidade de grãos de pólen de A. graveolens ingerida pelos adultos foi significativamente maior do que a dos polens de C. sativum e F. vulgare, em 24 horas, com o oposto ocorrendo em 48 horas. Nas primeiras 24 horas, os adultos ingeriram mais pólen de A. graveolens do que as larvas, ocorrendo o oposto com F. vulgare. Não houve diferença significativa na quantidade de pólen de Apiaceae ingerida entre as larvas e adultos em 48 horas. Os resultados sugerem que o hábito de alimentar de pólen de certas joaninhas afidófagas pode ser crucial para a sua preservação dentro dos agroecossistemas.

Foliar application of Zn at flowering stage improves plant’s performance, yield and yield attributes of black gram.

Black gram plants subjected to varying levels of Zn supply (0.01 to 10 µM Zn) showed optimum growth and dry matter yield in plants receiving 1 µM Zn. The dry matter yield of plants decreased in plants receiving 0.01 and 0.1 µM Zn (deficient) and excess levels of Zn (2 and 10 µM Zn). The plants grown with Zn deficient supply showed delayed flowering, premature bud abscission, reduced size of anthers, pollen producing capacity, pollen viability and stigma receptivity resulting in poor pod formation and seed yield. Providing Zn as a foliar spray at pre-flowering stage minimized the severity of Zn deficiency on reproductive structure development and enhanced the seed nutritional status by enhancing seed Zn density, seed carbohydrate (sugar and starch content) and storage proteins (albumins, globulins, glutenins, and prolamines).

Histological analysis of pollen-pistil interactions in sour passion fruit plants (Passiflora edulis Sims)

The success of sexual plant reproduction is directly influenced by specific interactions between the pollen and pistil. Light, fluorescence and scanning electron microscopy techniques were used to evaluate the steps of pollination in sour passion fruit plants (Passiflora edulis Sims). In the compatible interaction, pollen tubes grow through stigma projections towards the ovary. The pollen grain surface was found to be spheroidal and to consist of heteroreticulate exine with six colpi. Furthermore, analysis in vivo of pollen-pistil interactions indicated that stigmas of flowers 24 hours before anthesis are unable to discriminate compatible (genetically unrelated) and incompatible (genetically related) pollen grains. Taken together, these results provide insight into the cellular mechanisms underlying pollination in passion fruit plants.

Unión de los antibióticos tilosina, tilmicosina y oxitetraciclina a proteínas presentes en abejas, larvas y productos de la colmena de Apis mellifera L / Binding of tylosin, tilmicosin and oxytetracycline to proteins from honeybees, larvae and beehive products

Las abejas melíferas son afectadas por gran cantidad de enfermedades infecciosas principalmente producidas por bacterias, hongos, virus y parásitos eucariotas. Dentro de las ocasionadas por procariotas, la loque americana es una enfermedad extremadamente grave que afecta a larvas y pupas de abejas; su agente causal es la bacteria esporulada Paenibacillus larvae. La administración de antibióticos es la principal alternativa para el control de esta enfermedad en colmenares con altos niveles de infección. El objetivo del presente trabajo fue determinar, mediante un método biológico, la unión de los antibióticos tilosina, tilmicosina y oxitetraciclina a las proteínas presentes en abejas adultas, larvas menores de 72 horas, larvas mayores de 72 horas, jalea de obreras, miel y polen, con la finalidad de diseñar un modelo de ruta cinética de los antibióticos. Los límites de sensibilidad de la técnica de valoración de estos antibióticos fueron 0,05 μg/ml para tilosina y tilmicosina, y 0,01 μg/ml para oxitetraciclina. Los coeficientes de correlación fueron superiores a 0,90 y los coeficientes de variación intra e inter-ensayo inferiores al 5%. Tanto tilosina como oxitetraciclina presentaron un porcentaje de unión a proteínas de un 15% en promedio en tejidos y subproductos de la colmena, lo cual resultó inferior a lo observado con tilmicosina (29% en promedio). En conclusión, por sus características químicas, su actividad antimicrobiana y su baja tasa de unión a las abejas, larvas y subproductos de la colmena, la tilosina presenta propiedades farmacocinéticas que podrían representar una ventaja terapéutica para el tratamiento de la loque americana en colmenas.

American Foulbrood (AFB) caused by the spore-forming bacterium Paenibacillus larvae is the most serious disease of bacterial origin affecting larvae and pupae of honeybees. Antibiotics are used in many countries for the control of AFB in high incidence areas, but their misuse may lead to antibiotic resistance of bacterial strains and honey contamination. The objective of the present work was to determine, through a biological method, the protein binding of tylosin, tilmicosin and oxytetracycline to worker jelly; honey; pollen; adult bees and larvae in order to propose their kinetic routes. The sensitivity limit of the technique used was 0.05 μg/ml for tylosin and tilmicosin and 0.01 μg/ml for oxytetracycline, respectively. The method had intra and inter-assay correlation coefficients over 0.90, respectively and a coefficient variation of intra-and inter-assay for all antibiotics and processed samples under 5%. Tylosin and oxytetracycline presented lower percentages of protein binding in tissues and hive products (average 15%) in relation to those observed for tilmicosin (29%). In conclusion, tylosin is useful for AFB control in honey bee colonies due to its chemical characteristics, antimicrobial activity and levels of protein binding in bees, larvae, and beehive products.

Abnormal anther development and high sporopollenin synthesis in benzotriazole treated male sterile Helianthus annuus L.

Foliar application of 1.5% benzotriazole induced 100% pollen sterility in H. annuus. Pollen abortion in treated plants was mainly associated with abnormal behaviour of tapetum. A limited number of anther locule showed early degeneration of tapetum followed by disintegration of sporogenous tissues. On the other hand, some locules showed normal development of tapetum at initial stages. However, this tapetum exhibited degenerated and non-functional cell organelles. In both these situations tapetum failed to provide proper nourishment to developing microspores. The ultrastructure of both tapetum and microspores is different from that of control material with irregularities of exine deposition, endopolyploidy of tapetal nuclei and an alteration of organelle composition being correlated with sterility. Pollen grains thus developed were devoid of nucleus and cell organelles and were complete sterile.

Gene flow from transgenic to nontransgenic soybean plants in the Cerrado region of Brazil

Evaluation of transgenic crops under field conditions is a fundamental step for the production of genetically engineered varieties. In order to determine if there is pollen dispersal from transgenic to nontransgenic soybean plants, a field release experiment was conducted in the Cerrado region of Brazil. Nontransgenic plants were cultivated in plots surrounding Roundup Ready transgenic plants carrying the cp4 epsps gene, which confers herbicide tolerance against glyphosate herbicide, and pollen dispersal was evaluated by checking for the dominant gene. The percentage of cross-pollination was calculated as a fraction of herbicide-tolerant and -nontolerant plants. The greatest amount of transgenic pollen dispersion was observed in the first row, located at one meter from the central (transgenic) plot, with a 0.52% average frequency. The frequency of pollen dispersion decreased to 0.12% in row 2, reaching 0% when the plants were up to 10 m distance from the central plot. Under these conditions pollen flow was higher for a short distance. This fact suggests that the management necessary to avoid cross-pollination from transgenic to nontransgenic plants in the seed production fields should be similar to the procedures currently utilized to produce commercial seeds.

Selection and estimation of the heritability of sunflower (Helianthus annuus) pollen collection behavior in Apis mellifera colonies

We selected honey bee colonies (Apis mellifera L.) with a high tendency to collect sunflower pollen and estimated the heritability of this trait. The percentage of sunflower pollen collected by 74 colonies was evaluated. Five colonies that collected the highest percentages of sunflower pollen were selected. Nineteen colonies headed by daughters of these selected queens were evaluated for this characteristic in comparison with 20 control (unselected) colonies. The variation for the proportion of sunflower pollen was greater among colonies of the control group than among these selected daughter colonies. The estimated heritability was 0.26 +/- 0.23, demonstrating that selection to increase sunflower pollen collection is feasible. Such selected colonies could be used to improve sunflower pollination in commercial fields.

Ubisch bodies and pollen ontogeny in Oxalis articulata Savigny

The correlation between the ontogeny of Ubisch bodies and pollen development in Oxalis articulata was studied with Transmission Electron Microscopy (TEM). The ultrastructural changes observed during the different stages of development in the tapetal cells are related to Ubisch bodies, sporopollenin and pollen-kitt formation. The pro-orbicules have the appearance of lipid globuli and their formation is related to the endoplasmic reticulum of rough type (ERr). The lipid globules or pro-orbicules disappear in the mature Ubisch bodies, and the places that they occupied remain free of contents or with pollen-kitt.

Digestibilidad del polen de cactáceas columnares en los murciélagos glosofaginos Glossophaga longirostris y Leptonycteris curasoae (Chiroptera: Phyllostomidae) / Digestibility of columnar cacti pollen grains in the glosophagine bats Glossophaga longirostris and Leptonycteris curasoae (Chiroptera: Phyllostomidae)

We examined the protoplasmic assimilation of columnar cacti pollen grains in two species of Venezuelan desert glosophagine bats, Glossophaga longirostris and Leptonycteris curasoae, by determining the amount of empty (digested) pollen grains found in their fecal samples. To determine the amount of empty pollen grains, the fecal samples were stained to differ between empty and full (non-digested) pollen grains. The number of empty and full pollen grains observed in the fecal samples were corrected using the amount of aborted pollens present (before anthesis) in flowers of the columnar cacti species (Subpilocereus repandus, Stenocereus griseus and Pilosocereus tillianus ) used by bats as food in the study site; G. longirostris and L. curasoae digested 64.2% and 71.3% of all the pollens fed, respectively. These high values confirm the importance of pollen in the diet of these bats, given its high nitrogen level

Para estudiar la asimilación protoplasmática de los granos de polen de cactáceas columnares en dos especies de murciélagos glosofaginos, determinamos el porcentaje de granos de polen vacíos (digeridos) en muestras fecales de Glossophaga longirostris y Leptonycteris curasoae , capturados en una zona árida de Venezuela. Para determinar el porcentaje de granos de polen digeridos por los murciélagos, aplicamos un colorante sobre las muestras fecales que permite diferenciar sin dificultad el polen vacio del polen lleno. Después de cuantificar los granos de polen de ambas categorías en las muestras fecales, los valores fueron corregidos tomando en cuenta el porcentaje de polen vacío (previo a la antesis) en las flores maduras de las especies de cactáceas columnares (Subpilocereus repandus, Stenocereus griseus y Pilosocereus tillianus ). G. longirostris asimila el 64.2% de los granos de polen que ingiere y L. curasoae asimila el 71.3%. Estos elevados porcentajes confirman la importancia del polen en la dieta de estos murciélagos, dado su alto contenido de nitrógeno

Distribution of pectins in the pollen apertures of Oenothera hookeri: velans ster/+ster

Cell wall pectins are some of the most complex biopolymers known, and yet their functions remain largely mysterious. The aim of this paper was to deepen the study of the spatial pattern of pectin distribution in the aperture of Oenothera hookeri.velans ster/+ster fertile pollen. We used "in situ" immunocytochemical techniques at electron microscopy, involving monoclonal antibodies JIM5 and JIM7 directed against pectin epitopes in fertile pollen grains of Oenothera hookeri.velans ster/+ster. The same region was also analyzed by classical cytochemistry for polysaccharide detection. Immunogold labelling at the JIM7 epitope showed only in mature pollen labelling mainly located at the intine endo-aperture region. Cytoplasmic structures near the plasma membrane of the vegetative cell showed no labelling gold grains. In the same pollen stge the labelling at the JIM5 epitope was mostly confined to a layer located in the limit between the endexine and the ektexine at the level of the border of the oncus. Some tubuli at the base of the ektexine showed also an accumulation of gold particles. No JIM5 label was demonstrated in the aperture chamber and either in any cytoplasmic structure of the pollen grains. The immunocytochemical technique, when compared with the traditional methods for non-cellulose polysaccharide cytochemistry is fare more sensitive and allows the univocal determination of temporal and spatial location of pectins recognized by the JIM7 and JIM5 MAbs.