Flavonoids, phenolic acids and abscisic acid in Australian and New Zealand Leptospermum honeys

Flavonoids, phenolic acids and abscisic acid of Australian and New Zealand Leptospermum honeys were analyzed by HPLC. Fifteen flavonoids were isolated in Australian jelly bush honey (Leptospermum polygalifolium), with an average content of 2.22 mg/100 g honey. Myricetin (3,5,7,3',4',5'-hexahydroxyflavone), luteolin (5,7,3',4'-tetrahydroxyflavone) and tricetin (5,7,3',4',5'-pentahydroxyflavone) were the main flavonoids identified. The mean content of total phenolic acids in jelly bush honey was 5.14 mg/100 g honey, with gallic and coumaric acids as the potential phenolic acids. Abscisic acid was quantified as twice the amount (11.6 mg/100 g honey) of the phenolic acids in this honey. The flavonoid profile mainly consisted of quercetin (3,5,7,3',4'-pentahydroxyflavone), isorhamnetin (3,5,7,4'-tetrahydroxyflavone 3'-methyl ethyl), chrysin (5,7-dihydroxyflavone), luteolin and an unknown flavanone in New Zealand manuka (Leptospermum scoparium) honey with an average content of total flavonoids of 3.06 mg/100 g honey. The content of total phenolic acids was up to 14.0 mg/100 g honey, with gallic acid as the main component. A substantial quantity (32.8 mg/100 g honey) of abscisic acid was present in manuka honey. These results showed that flavonoids and phenolic acids could be used for authenticating honey floral origins, and abscisic acid may aid in this authentication. (C) 2002 Published by Elsevier Science Ltd.

Isolamento e identificação da morina em mel brasileiro de Apis mellifera

The flavonoid fraction was purified by a combination of chromatography on Amberlite XAD-2 and preparative silica gel TLC. Morin (3, 5, 7, 2', 4'- pentahydroxyflavone) was the only flavonol found in honey from Brazilian Citrus sp.. The structure of morin was determined on the basis of UV and ¹H and 13C NMR spectral data together with literature references. This is the first report on the isolation of morin from Brazilian Citrus honey.

Phenolic acids and abscisic acid in Australian Eucalyptus honeys and their potential for floral authentication

Seven phenolic acids related to the botanical origins of nine monofloral Eucalyptus honeys from Australia, along with two abscisic isomers, have been analyzed. The mean content of total phenolic acids ranges from 2.14 mg/100 g honey of black box (Eucalyptus largiflorens) honey to 10.3 mg/100 g honey of bloodwood (Eucalyptus intermedia) honey, confirming an early finding that species-specific differences of phytochemical compositions occur quantitatively among these Eucalyptus honeys. A common profile of phenolic acids, comprising gallic, chlorogenic, coumaric and caffeic acids, can be found in all the Eucalyptus honeys, which could be floral markers for Australian Eucalyptus honeys. Thus, the analysis of phenolic acids could also be used as an objective method for the authentication of botanical origin of Eucalyptus honeys. Moreover, all the honey samples analyzed in this study contain gallic acid as the main phenolic acid, except for stringybox (Eucalyptus globoidia) honey which has ellagic acid as the main phenolic acid. This result indicates that the species-specific differences can also be found in the honey profiles of phenolic acids. Further-more, the analysis of abscisic acid in honey shows that the content of abscisic acid varies from 0.55 mg/100 g honey of black box honey to 4.68 mg/ 100 g honey of bloodwood honey, corresponding to the contents of phenolic acids measured in these honeys. These results have further revealed that the HPLC analysis of honey phytochemical constituents could be used individually and/or jointly for the authentication of the botanical origins of Australian Eucalyptus honeys. (C) 2003 Elsevier Ltd. All rights reserved.

Flavonoids in Australian Melaleuca, Guioa, Lophostemon, Banksia and Helianthus honeys and their potential for floral authentication

Flavonoids in Australian honeys from five botanical species (Melaleuca, Guioa, Lophostemon, Banksia and Helianthus) have been analyzed in relation to their floral origins. Tea tree (Melaleuca quinquenervia) and heath (Banksia ericifolia) honeys show a common flavonoid profile comprising myricetin (3,5,7,3',4',5'-hexahydroxyflavone), tricetin (5,7,3',4,5'-pentahydroxyflavone), querectin (3,5,7,3',4'-pentahydroxyflavone) and luteolin (5,7,3',4'-tetrahydroxyflavone), which was previously suggested as a floral marker for an Australian Eucalyptus honey (bloodwood or Eucalyptus intermedia honey). These honeys of various floral species can be differentiated by their levels of total flavonoids, being 2.12 mg/100 g for heath honey and 6.35 m/100 g for tea tree honey. In brush box (Lophostemon conferta) honey, the flavonoid profile comprising mainly tricetin, luteolin and quercetin is similar to that of another Eucalyptus honey (yellow box or Eucalyptus melliodora honey). These results indicate that the flavonoid profiles in some of the Australian non-Eucalyptus honeys may contain more or less certain flavonoids from Eucalyptus floral sources because of the diversity and extensive availability of Eucalyptus nectars for honeybee foraging yearly around or a possible cross contamination of the monofloral honeys during collection, transportation and/or storage. Further analyses are required to differentiate and/or verify the botanical sources of the flavonoids that contribute to the flavonoid profiles of these honeys, by restricting honey sampling areas and procedures, employing other complementary analytical methods (e.g. pollen analysis, sugar profile) and using materials (e.g. nectar) directly sourced from the flowering plant for comparative studies. In Australian crow ash (Guioa semiglauca) honey, myricetin, tricetin, quercetin, luteolin and an unknown flavonoid have been found to be the main flavonoids, which is characteristic only to this type of honey, and could thus be used as the floral marker, while in Australian sunflower (Helianthus annuus) honey, the content of total flavonoids is the smallest amount comparing to those in the other honeys analysed in this study. However, the flavonoid quercetin and the flavonoid profile mainly consisting of quercetin, quercetin 3,3'-dimethyl ether (5,7,4'-trihydroxy3,3'-dimethoxyflavone), myricetin and luteolin are characteristic only to this sunflower honey and could thus be used for the authentication.

Phenolic acids in Australian Melaleuca, Guioa, Lophostemon, Banksia and Helianthus honeys and their potential for floral authentication

Eight phenolic acids and two abscisic acid isomers in Australian honeys from five botanical species (Melaleuca, Guioa, Lophostemon, Banksia and Helianthus) have been analyzed in relation to their botanical origins. Total phenolic acids present in these honeys range from 2.13 mg/100 g sunflower (Helianthus annuus) honey to 12.11 mg/100 g tea tree (Melaleuca quinquenervia) honey, with amounts of individual acids being various. Tea tree honey shows a phenolic profile of gallic, ellagic, chlorogenic and coumaric acids, which is similar to the phenolic profile of an Australian Eucalyptus honey (bloodwood or Eucalyptus intermedia honey). The main difference between tea tree and bloodwood honeys is the contribution of chlorogenic acid to their total phenolic profiles. In Australian crow ash (Guioa semiglauca) honey, a characteristic phenolic profile mainly consisting of gallic acid and abscisic acid could be used as the floral marker. In brush box (Lophostemon conferta) honey, the phenolic profile, comprising mainly gallic acid and ellagic acid, could be used to differentiate this honey not only from the other Australian non-Eucalyptus honeys but also from a Eucalyptus honey (yellow box or Eucalyptus melliodora honey). However, this Eucalyptus honey could not be differentiated from brush box honey based only on their flavonoid profiles. Similarly, the phenolic profile of heath (Banksia ericifolia) honey, comprising mainly gallic acid, an unknown phenolic acid (Phl) and coumaric acid, could also be used to differentiate this honey from tea tree and bloodwood honeys, which have similar flavonoid profiles. Coumaric acid is a principal phenolic acid in Australian sunflower honey and it could thus be used together with gallic acid for the authentication. These results show that the HPLC analysis of phenolic acids and abscisic acids in Australian floral honeys Could assist the differentiation and authentication of the honeys. © 2005 Elsevier Ltd. All rights reserved.

Polinização em cultura de laranja (Citrus sinensis L. Osbeck, var. Pera-Rio)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genome Sequence Of Streptomyces Wadayamensis Strain A23, An Endophytic Actinobacterium From Citrus Reticulata.

The actinobacterium Streptomyces wadayamensis A23 is an endophyte of Citrus reticulata that produces the antimycin and mannopeptimycin antibiotics, among others. The strain has the capability to inhibit Xylella fastidiosa growth. The draft genome of S. wadayamensis A23 has ~7.0 Mb and 6,006 protein-coding sequences, with a 73.5% G+C content.

Africanized honey bees are efficient at detecting, uncapping and removing dead brood

The hygienic behavior of honey bees is based on a two-step process, including uncapping and removing diseased, dead, damaged, or parasitized brood inside the cell. We evaluated during periods of 1 h the time that hygienic and non-hygienic colonies of Africanized honey bees spend to detect, uncap and remove pin-killed brood using comb inserts with transparent walls placed in observation hives. We observed that hygienic colonies are significantly faster in detecting, uncapping and removing dead brood in the cells (P < 0.001).

A scientific note about spectroscopic analysis of honey bee brood comb cappings in hygienic and non-hygienic honey bee colonies

Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)

Enzymatic differences between the endophyte Guignardia mangiferae (Botryosphaeriaceae) and the citrus pathogen G. citricarpa

The endophyte Guignardia mangiferae is closely related to G. citricarpa, the causal agent of citrus black spot; for many years these species had been confused with each other. The development of molecular analytical methods has allowed differentiation of the pathogen G. citricarpa from the endophyte G. mangiferae, but the physiological traits associated with pathogenicity were not described. We examined genetic and enzymatic characteristics of Guignardia spp strains; G. citricarpa produces significantly greater amounts of amylases, endoglucanases and pectinases, compared to G. mangiferae, suggesting that these enzymes could be key in the development of citrus black spot. Principal component analysis revealed pectinase production as the main enzymatic characteristic that distinguishes these Guignardia species. We quantified the activities of pectin lyase, pectin methylesterase and endopolygalacturonase; G. citricarpa and G. mangiferae were found to have significantly different pectin lyase and endopolygalacturonase activities. The pathogen G. citricarpa is more effective in pectin degradation. We concluded that there are significant physiological differences between the species G. citricarpa and G. mangiferae that could be associated with differences in pathogenicity for citrus plants.

The four hexamerin genes in the honey bee: structure, molecular evolution and function deduced from expression patterns in queens, workers and drones

Background: Hexamerins are hemocyanin-derived proteins that have lost the ability to bind copper ions and transport oxygen; instead, they became storage proteins. The current study aimed to broaden our knowledge on the hexamerin genes found in the honey bee genome by exploring their structural characteristics, expression profiles, evolution, and functions in the life cycle of workers, drones and queens. Results: The hexamerin genes of the honey bee (hex 70a, hex 70b, hex 70c and hex 110) diverge considerably in structure, so that the overall amino acid identity shared among their deduced protein subunits varies from 30 to 42%. Bioinformatics search for motifs in the respective upstream control regions (UCRs) revealed six overrepresented motifs including a potential binding site for Ultraspiracle (Usp), a target of juvenile hormone (JH). The expression of these genes was induced by topical application of JH on worker larvae. The four genes are highly transcribed by the larval fat body, although with significant differences in transcript levels, but only hex 110 and hex 70a are re-induced in the adult fat body in a caste-and sex-specific fashion, workers showing the highest expression. Transcripts for hex 110, hex 70a and hex70b were detected in developing ovaries and testes, and hex 110 was highly transcribed in the ovaries of egg-laying queens. A phylogenetic analysis revealed that HEX 110 is located at the most basal position among the holometabola hexamerins, and like HEX 70a and HEX 70c, it shares potential orthology relationship with hexamerins from other hymenopteran species. Conclusions: Striking differences were found in the structure and developmental expression of the four hexamerin genes in the honey bee. The presence of a potential binding site for Usp in the respective 5' UCRs, and the results of experiments on JH level manipulation in vivo support the hypothesis of regulation by JH. Transcript levels and patterns in the fat body and gonads suggest that, in addition to their primary role in supplying amino acids for metamorphosis, hexamerins serve as storage proteins for gonad development, egg production, and to support foraging activity. A phylogenetic analysis including the four deduced hexamerins and related proteins revealed a complex pattern of evolution, with independent radiation in insect orders.

Sequential hygienic behavior in Carniolan honey bees (Apis mellifera carnica)

We examined the sequence, order or steps of hygienic behavior (HB) from pin-killed pupae until the removal of them by the bees. We conducted our study with four colonies of Apis mellifera carnica in Germany and made four repetitions. The pin-killing method was used for evaluation of the HB of bees. The data were collected every 2 h after perforation, totaling 13 observations. Additionally, for one hygienic colony and another non-hygienic colony, individual analyses of each dead pupa were made at every observation, including all details, steps or sequences of HB. The bees recognize the cells containing dead pupae within 2 h after perforation, initially making a hole in the capping, which is the beginning of HB. Uncapping of the dead brood cell reached maximum values from 4 to 6 h after perforation; after 24 h, practically all cells were already uncapped. Another variable, called brood partially removed, was analyzed 4 h after perforation, after the cells had been perforated, which involved uncapping, followed by partial or total removal of the brood. Maximum values of brood partially removed were found 10 h after perforation, though such cells could be found up to 48 h after perforation. The most frequent sequence of events in both colonies was: capped cell -> punctured cell. brood partially removed -> empty cell. A new model of three pairs of recessive genes (uncapping u1, u2 and remover r) was proposed in order to explain the genetic control of the HB in Apis mellifera. We recommend evaluating HB 24 h after perforation and using a correction factor to compensate for control removal levels. We found a series of details of HB, which allow a study of how various factors may affect the sequence of the activities involved in HB and investigation of the genetics that controls this process.

Comparative study of the hygienic behavior of Carniolan and Africanized honey bees directed towards grouped versus isolated dead brood cells

In Apis mellifera, hygienic behavior involves recognition and removal of sick, damaged or dead brood from capped cells. We investigated whether bees react in the same way to grouped versus isolated damaged capped brood cells. Three colonies of wild-type Africanized honey bees and three colonies of Carniolan honey bees were used for this investigation. Capped worker brood cells aged 12 to 14 days old were perforated with the pin-killing method. After making holes in the brood cells, the combs were placed back into the hives; 24 h later the number of cleaned cells was recorded in areas with pin-killed and control brood cells. Four repetitions were made in each colony. Isolated cells were more frequently cleaned than grouped cells, though variance analysis showed no significant difference (P = 0.1421). Carniolan bees also were somewhat, though not significantly more hygienic than Africanized honey bees (P = 0.0840). We conclude that honey bees can detect and remove both isolated and grouped dead brood. The tendency towards greater hygienic efficiency directed towards grouped pin-killed brood may be a consequence of a greater concentration of volatiles emanating from the wounds in the dead pupae.

Pollination of tomatoes by the stingless bee Melipona quadrifasciata and the honey bee Apis mellifera (Hymenoptera, Apidae)

The pollination effectiveness of the stingless bee Melipona quadrifasciata and the honey bee Apis mellifera was tested in tomato plots. The experiment was conducted in four greenhouses as well as in an external open plot in Ribeirao Preto, SP, Brazil. The tomato plants were exposed to visits by M. quadrifasciata in one greenhouse and to A. mellifera in another; two greenhouses were maintained without bees (controls) and an open field plot was exposed to pollinators in an area where both honey bee and stingless bee colonies are abundant. We counted the number of tomatoes produced in each plot. Two hundred tomatoes from each plot were weighed, their vertical and transversal circumferences were measured, and the seeds were counted. We collected 253 Chrysomelidae, 17 Halictidae, one Paratrigona sp, and one honey bee from the flowers of the tomato plants in the open area. The largest number of fruits (1414 tomatoes), the heaviest and largest tomatoes, and the ones with the most seed were collected from the greenhouse with stingless bees. Fruits cultivated in the greenhouse with honey bees had the same weight and size as those produced in one of the control greenhouses. The stingless bee, M. quadrifasciata, was significantly more efficient than honey bees in pollinating greenhouse tomatoes.

Changes in integument structure during the imaginal molt of the honey bee

The changing pattern of developing cuticle and associated epidermis is described during the imaginal molt in the honey bee. Observations began immediately after the pupal molt, and included histological analyses of the integument during apolysis and the subsequent deposition and differentiation of the adult cuticle. Apolysis coincides with a marked increase in the thickness and reorganization of the epidermal layer, reflecting changes in cell structure. The epidermis remains thickened during the period of cuticle deposition, suggesting intense biosynthetic activity, but turns into a very thin layer during cuticle differentiation, clearly indicating that secretory activity for cuticle formation is terminating. The thoracic cuticle differentiates earlier and becomes thicker than the abdominal. The observed changes in integument structure provide insights that permit an improved physiological characterization for staging pupal and pharate adult development.