Effect of different drying methods on the amino acids,α-dicarbonyls and volatile compounds of rape bee pollen

The significant demand for high quality food has motivated us to adopt appropriate processing methods to improve the food nutritional quality and flavors.In this study,the effects of five drying methods,namely,pulsed vacuum drying(PVD),freeze drying(FD),infrared drying(IRD),hot-air drying(HAD)and sun drying(SD)on free amino acids(FAAs),α-dicarbonyl compounds(α-DCs)and volatile compounds(VOCs)in rape bee pollen(RBP)were determined.The results showed that FD significantly released the essential amino acids(EAAs)compared with fresh samples while SD caused the highest loss.Glucosone was the dominantα-DCs in RBP and the highest loss was observed after PVD.Aldehydes were the dominant volatiles of RBP and SD samples contained more new volatile substances(especially aldehydes)than the other four drying methods.Comprehensively,FD and PVD would be potential methods to effectively reduce the quality deterioration of RBP in the drying process.

Inhibitory effect of procyanidin B2 and tannin acid on cholesterol esterase and their synergistic effect with orlistat

This study was aimed to analyze the effect of procyanidin B2(PC)and tannin acid(TA)on the activities of cholesterol esterase(CEase)and the inhibitory mechanisms of enzymatic activity.The interaction mechanisms were investigated by enzymatic kinetics,multi-spectroscopy methods,thermodynamics analysis,molecular docking,and dynamic simulations.PC and TA could bind with CEase and inhibit the activity of enzyme in a mixed-competitive manner and non-competitive manner,which was verified by molecular docking simulations and dynamics simulations.Also,PC and TA showed the synergistic inhibition with orlistat.Fluorescence,UVvis and the thermodynamic analysis revealed that the complexes were formed from CEase and inhibitors by noncovalent interaction.As revealed by the circular dichroism results,both PC and TA decreased enzymatic activities by altering the conformations of CEase.The inhibition of PC and TA on CEase might be one mechanism for its cholesterol-lowering effect.

Functional characterization of sensory neuron membrane protein 1a involved in sex pheromone detection of Apolygus lucorum(Hemiptera:Miridae)

The mirid bug Apolygus lucorum(Hemiptera:Miridae)is a polyphagous pest that affects a wide range of host plants.Its control remains challenging mainly due to its rapid reproduction,necessitating an understanding of sex pheromone communication.The recognition of sex pheromones is vital for courtship and mating behaviors,and is mediated by various chemosensory-associated proteins.Among these,sensory neuron membrane protein(SNMP),a CD36-related protein,is suggested to play crucial roles in detecting sex pheromones.In this study,we employed transcriptomic and genomic data from A.lucorum and phylogenetic approaches,and identified four putative SNMP genes(AlucSNMP1a,AlucSNMP1b,AlucSNMP2a,and AlucSNMP2b)with full open reading frames.Expression analysis revealed the ubiquitous presence of AlucSNMP transcripts in multiple tissues,with only AlucSNMP1a exhibiting male-biased expression in the antennae,suggesting its potential role in male chemosensation.Functional analysis using the Xenopus oocyte expression system,coupled with two-electrode voltage clamp recording,demonstrated that the co-expression of AlucSNMP1a with specific pheromone receptors(PRs)and the Odorant receptor co-receptor(Orco)significantly enhanced electrophysiological responses to sex pheromones compared to the co-expression of PRs and Orco alone.Moreover,the results indicated that the presence of AlucSNMP1a not only affected the responsiveness to sex pheromones but also influenced the kinetics(activation and inactivation)of the induced signals.In contrast,the co-expression of AlucSNMP1b with AlucPR/Orco complexes had no impact on the inward currents induced by two pheromone compounds.An examination of the selective pressures on SNMP1 genes across 20 species indicated strong purifying selection,implying potential functional conservation in various insects.These findings highlight the crucial role of AlucSNMP1a in the response to sex pheromones.

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

Characterization of physicochemical and immunogenic properties of allergenic proteins altered by food processing:a review

Food allergens are mainly naturally-occurring proteins with immunoglobulin E(IgE)-binding epitopes.Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether and how food processing techniques reduce allergenicity.We here discuss the impacts of food processing technologies on the modification of physicochemical,structural,and immunogenic properties of allergenic proteins.Detection techniques for characterizing changes in these properties of food allergens are summarized.Food processing helps to reduce allergenicity by aggregating or denaturing proteins,which masks,modifies,or destroys antigenic epitopes,whereas,it cannot eliminate allergenicity completely,and sometimes even improves allergenicity by exposing new epitopes.Moreover,most food processing techniques have been tested on purified food allergens rather than food products due to potential interference of other food components.We provide guidance for further development of processing operations that can decrease the allergenicity of allergenic food proteins without negatively impacting the nutritional profile.

Reducing pesticide use:Synthesis and application of ROS-SPC as an efficient nanocarrier and scavenger of reactive oxygen species in plants

There is an increasing need to reduce the use of pesticides to reduce their potential threat to food/environmental safety.At the same time,an increase in reactive oxygen species(ROS)induced by abiotic stresses in plants can lead to an increase in ROS in the plant and affect yield.In this paper,ROS-SPC was synthesised by two reactions and used as an efficient pesticide nanocarrier/adjuvant and scavenger of reactive oxygen species(ROS)in plants.By hydrophobic interaction,ROS-SPC spontaneously conjugated tofluazinam with a pesticide loading capacity(PLC)of 15.1%.Afterfluazinam was conjugated to ROS-SPC,the particle size offluazinam was reduced from 64.70 nm reduced to 19.82 nm,and the contact angle of pesticide droplets on plant leaves was significantly reduced from 59.44°to 26.76°.ROS-SPC as a carrier was tested to inhibit phytopathogenic fungi by 200%more than conventional delivery methods.In addition,we also learned that ROS-SPC with endocytosis capability can indeed remove reactive oxygen species from plants.Tests using HUVEC cells showed that ROS-SPC has low cytotoxicity within a reasonable range of applications,and ROS-SPC was tested to have low toxicity to pollinating bees.

The Influence of Bt-Transgenic Maize Pollen on the Bacterial Diversity in the Midgut of Chinese Honeybees, Apis cerana cerana

Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene （800 ng mL^-1） and supercoiled plasmid DNA （800 ng mL^-1） under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon＇s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.

Effect of fermented bee pollen on metabolic syndrome in high-fat diet-induced mice

Bee pollen has potential in preventing metabolic syndrome(MetS).The present study aimed to investigate the effect of yeast-fermented wall-broken bee pollen(YB)intervention on ICR mice with MetS induced with a high-fat(HF)diet.After YB intervention in mice for 16 weeks,the effect on alleviating MetS was evaluated based on MetS serum parameters,hepatic oxidant status markers and gut microbial populations.The results of animal experiment showed that YB intervention attenuated MetS.Based on multivariate statistical analysis results,YB treatment signifi cantly increased glutathione S-transferase(GST)and catalase(CAT)activities and decreased the malondialdehyde(MDA)level in the liver.Further investigation showed that YB restored the Nrf-2-Keap-1 pathway to alleviate oxidative stress.Additionally,gut microbial community analysis revealed that YB restored the increase in the Firmicutes to Bacteroidetes(F/B)ratio(6.94 for the HF group and 3.74 for HF+YB group)and improved Lactobacillus and Lactococcus abundance induced by the HF diet.Overall,YB improved function and prevented MetS by modulating the gut microbiota and alleviating oxidative stress.

Comparative Analyses of Proteome Complement Between Worker Bee Larvae of High Royal Jelly Producing Bees (A. m. ligustica) and Carniolian Bees (A. m. carnica)

This study is to compare the protein composition of the high royal jelly producing bee （A. m. ligustica） with that of Carniolian bee （A. m. carnica） during their worker larval developmental stage. The experiment was carried out by two- dimensional gel electrophoresis. The results showed that significant higher numbers of total proteins （283） were detected in larvae of high royal jelly producing bees （Jelly bee） than those of Carniolian bees （152） on 2-d-old larvae. Among them, 110 proteins were presented on both strains of bee larvae, whereas 173 proteins were specific to larvae of Jelly bees, and 42 proteins were exclusive to Carniolian larvae. However, on the 4th d, a significant higher number of total proteins （290） were detected in larvae of Jelly bees than those of Carniolian bees （240）, 163 proteins resolved to both bee larvae, and 127 proteins were specific to Jelly bees and 77 proteins to Camiolian bees. Until the 6th d, also a significant higher number of total proteins （236） were detected in larvae of Jelly bees than those of Carniolian bees （180）, 132 proteins were constantly expressed in two bee larvae, whereas 104 and 48 proteins are unique to Jelly bee and Carniolian bee larvae, respectively. We tentatively concluded that the metabolic rate and gene expression of Jelly bees larvae is higher than those of Carniolian bees based proteins detected as total proteins and proteins specific to each stage of two strains of bee larvae. Proteins constantly expressed on 3 stages of larval development with some significant differences between two bee strains, and proteins unique to each stage expressed differences in term of quality and quantity, indicating that larval development needed house keeping and specific proteins to regulate its growth at different development phage, but the expression mold is different between two strains of larval development.

Profile Analysis of the Proteome of the Egg of the High Royal Jelly Producing Bees (Apis mellifera L.)

The protein composition of the egg development in the high royal jelly producing bees （Apis mellifera L.） was investigated. This pioneer study was to separate and quantify the proteins in the egg of the high royal jelly producing worker bees （Apis mellifera L.） by using two-dimensional gel electrophoresis along with their three-day development. The results showed that 160, 195, and 176 proteins, with a wide range of molecular weight （17-80 KDa） and relatively narrow scope of pI （4. 00-8.40） could be detected on day 1, day 2, and day 3, respectively, during the developmental process of the egg. Meanwhile 44 protein spots were constantly detected along with the egg development. Among them 36% were in the uptrend along with the egg development, 14% were in the downtrend, and 39% were of the largest expressed volume on day 2. In addition, the specific proteins were expressed on day 1, day 2, and day 3 （89, 77, and 80, respectively）. Besides the coexistent and specific proteins, 24 proteins were expressed on day 1 and day 2, but silenced on day 3, 49 proteins were expressed on day 2 and day 3, but silenced on day 1, only 3 proteins were expressed on day 1 and day 3, but silenced on day 2. The result indicates that egg development is a sequential and complex gene controlled process, where the eggs of day 2 express the most active proteins. The coexistent proteins suggest that it is conservative and indispensable for this event. These specific proteins suggest that the different developmental stage needs specific proteins to regulate it.

Expression of genes involved in progesterone receptor paracrine signaling and their effect on litter size in pigs

Background： Embryonic mortality during the period of implantation strongly affects litter size in pigs. Progesterone receptor（PGR） paracrine signaling has been recognized to play a significant role in embryonic implantation. IHH,NR2F2, BMP2, FKBP4 and HAND2 were proved to involve in PGR paracrine signaling. The objective of this study was to evaluate the expression of IHH, NR2F2, BMP2, FKBP4 and HAND2 in endometrium of pregnant sows and to further investigate these genes＇ effect on litter size in pigs. Real-time PCR, western blot and immunostaining were used to study target genes/proteins expression in endometrium in pigs. RFLP-PCR was used to detect single nucleotide polymorphisms（SNPs） of target genes.Results： The results showed that the m RNA and protein expression levels of IHH, NR2F2 and BMP2 were up-regulated during implantation period（P 〈 0.05 or P 〈 0.01）. All target proteins were mainly observed in luminal epithelium and glandular epithelium. Interestingly, the staining of NR2F2 and HAND2 was also strong in stroma. SNPs detection revealed that there was a-204 C 〉 A mutation in promoter region of NR2F2 gene. Three genotypes were found in Large White, Landrace and Duroc sows. A total of 1847 litter records from 625 sows genotyped at NR2F2 gene were used to analyze the total number born（TNB） and number born alive（NBA）. The study of the effect on litter size suggested that sows with genotype CC tend to have higher litter size.Conclusions： These results showed the expression patterns of genes/proteins involved in PGR paracrine signaling over implantation time. And the candidate gene for litter size was identified from genes involved in this signaling. This study could be a resource for further studies to identify the roles of these genes for embryonic implantation in pigs.

Overexpression of a Glycosyltransferase Gene from a Metabolically Poly-Resistant Beckmannia syzigachne Population Alters Growth and Confers Herbicide Resistance to Brachypodium distachyon

Beckmannia syzigachne is a noxious weed for rice-wheat rotations in China.The B.syzigachne(AH-02)population evolved metabolic resistance to fenoxaprop-P-ethyl and mesosulfuron-methyl.To investigate the function of GT73C1 in this population,the GT73C1 gene was amplified by reverse transcription-polymerase chain reaction,and the sequence was 100%consistent with the transcriptome data.Its phylogenetic tree was displayed and annotated using FigTree v1.4.4.The plant overexpression vector of GT73C1 gene was constructed and used to transform Brachypodium distachyon plants.Furthermore,the expression of GT73C1 was significantly induced by fenoxaprop-P-ethyl and mesosulfuron-methyl,which was consistent with the findings from the whole plant bioassay.These results indicate that GT73C1 is closely related to the metabolic resistance of B.distachyon.

Possibility of Socio-economic Attributes and ICT Application in Improving Quality of Farmers

Evaluation on the diversified Information and Communication Technologies( ICTs) application along their association in socio-economic attributes can provide valuable insights about the trends of farmers towards improvement.Therefore,analytical research design and descriptive statistics were used to analyze the data collected from 400 respondents within different regions of Punjab Province in Pakistan.Based on the findings,it is recommended that government in collaboration with all stakeholders should launch regular,holistic and modern ICTs programs like applications of robo calls,SMS,etc.services at central level to ensure timely availability of agricultural information to improving the quality of young farmers.Furthermore,it is recommended to launch effective monitoring and evaluation mechanism of ICTs for agricultural development.

Transcriptome profiling during double-flower development provides insight into stamen petaloid in cultivated Lilium

Asiatic hybrid lilies(Lilium spp.),as the biggest cultivar groups of the lily variety are an ornamental plant with elegant floral patterns and bright colors.We discovered a double-flower Asiatic hybrid cultivar‘Annemarie’s Dream’whose stamens convert into petaloid stamens in varying degrees.Double-flower is a significant ornamental trait of the flower organ.However,the molecular mechanism of stamen petaloid formation has not been widely studied in Asiatic hybrid lily.Therefore,we used RNA-seq to contrast transcriptomes of stamen,petaloid stamen and inner tepal at two developmental stages.In total,190,488 unigenes were obtained and 37,549 differentially expressed genes(DEGs)were identified.We focused on DEGs involved in phytohormone signaling and transcription factor regulation,especially the MADS-box genes(A-class gene LiAP1;B-class gene LiPI;C-class gene LiAG;E-class gene LiAGL6,LiSEP3).Furthermore,we performed weight gene co-expression network analysis(WGCNA)and identified two co-expressed MADS-box homeotic genes(LiAG,CL14315.Contig2_All;LiAP1,CL10060.Contig3_All)as hubs.We also found that CL3014.Contig2_All(PYL)and CL5627.Contig1_All(GID2)as phytohormone-related genes may participate in the regulation of the stamen petaloid during double-flower development.In summary,our findings provide an insight into the molecular regulatory network underlying stamen petaloid and thereby offering a theoretical basis for double-flower breeding in Lilium.

Biochemistry and transcriptome analysis reveal condensed tannins alleviate liver injury induced by regulating cholesterol metabolism pathway

Free cholesterol has been considered to be a critical risk factor of nonalcoholic fatty liver disease(NAFLD).It remains unknown whether dietary intake of condensed tannins(CTs)have distinguishable effects to alleviate liver damage caused by a high cholesterol diet.Male C57BL/6 mice were fed a high cholesterol diet for 6 weeks,and given CTs treatment at a dosage of 200 mg/(kg·day)at the same time.The results indicated that compared with mice fed a normal diet,a high cholesterol diet group resulted in significant weight loss,dysregulation of lipid metabolism in blood and liver,and oxidative stress in the liver,but CTs treatment dramatically reversed these negative effects.Hematoxylin and eosin(H&E)staining and frozen section observation manifested that CTs treatment could effectively reduce the deposition of liver cholesterol and tissue necrosis caused by high cholesterol intake.CTs alleviated liver injury mainly by regulating the expression of related genes in cholesterol metabolism pathway and AMPK phosphorylation.Our results confirmed that CTs have remarkable cholesterol lowering and anti-liver injury effects in vivo.

Differential volatile organic compounds in royal jelly associated with different nectar plants

The aim of this work was to distinguish volatile organic compound（VOC） profiles of royal jelly（RJ） from different nectar plants. Headspace solid-phase microextraction（HS-SPME） was used to extract VOCs from raw RJ harvested from 10 nectar plants in flowering seasons. Qualitative and semi-quantitative analysis of VOCs extracts were performed by gas chromatography-mass spectrometry（GC-MS）. Results showed that VOC profiles of RJ from the samples were rich in acid, ester and aldehyde compound classes, however, contents of them were differential, exemplified by the data from acetic acid, benzoic acid methyl ester, hexanoic acid and octanoic acid. As a conclusion, these four VOCs can be used for distinguishing RJ harvested in the seasons of different nectar plants.

Research progress on distribution,sources,identification,toxicity,and biodegradation of microplastics in the ocean,freshwater,and soil environment

Microplastics(MPs)are distributed in the oceans,freshwater,and soil environment and have become major pollutants.MPs are generally referred to as plastic particles less than 5 mm in diameter.They consist of primary microplastics synthesized in microscopic size manufactured production and secondary microplastics generated by physical and environmental degradation.Plastic particles are long-lived pollutants that are highly resistant to environmental degradation.In this review,the distribution and possible sources of MPs in aquatic and terrestrial environments are described.Moreover,the adverse effects of MPs on natural creatures due to ingestion have been discussed.We also have summarized identification methods based on MPs particle size and chemical bond.To control the pollution of MPs,the biodegradation of MPs under the action of different microbes has also been reviewed in this work.This review will contribute to a better understanding of MPs pollution in the environment,as well as their identification,toxicity,and biodegradation in the ocean,freshwater,and soil,and the assessment and control of microplastics exposure.

Breeding the Mite-Resistant Honeybee by Nutritional Crossbreed Technology

Mite （Varroa destructor） is one of the most serious parasite threats to the honey bee （Apis mellifera） reared in China. The beekeepers mainly use the drug to control and kill the mite in the past years, but the honey products may be contaminated and the mite is becoming drug-resistant. The main idea of this paper is to research the possibility of mite-resistant honeybee rearing by nutritional crossbreed. The larvae （Apis mellifera ligustica） are bred with the royal jelly of Apis carana carana, and then the morphological index of the worker generation, genotypic frequency and gene frequency of the MDH Ⅱ, genetic resemblance, and mite resistance are measured. The results show that： compared to the parent workers, the proboscis length, anterior wing area, the total length of the third and fourth dorsal plate of the abdomen, the length of the fourth dorsal plate of the tuberculum, the area of the sixth abdominal segment, and the area of wax mirrors are significantly different, but the differences in the brachium index, dactylus index, and wing claw are not significant. Moreover, there are some mutations in the genotypic frequency and gene frequency of the MDH Ⅱ. The mite resistance of the nutritional crossbreed worker is significantly higher. The morphological, physical, and biochemical characters, genetic resemblance, and the mite-resistant ability of the worker generation can be changed by nutritional crossbreeding. Nutritional crossbreeding can be a new way to breed the honeybee.

Non-associative learning underlies pollination interaction of pollinators and flowering plants

The behavioral response of pollinators is significantly influenced by the prior experience of flower visiting.Learning of pollinators,including non-associative learning,associative learning,and operant conditioning,is determined by the presence or absence of rewards during the flower visiting experience.Here,we indicate that process of non-rewarding flower(empty flower)visiting coincident well with the behavioral paradigm of non-associative learning.Habituation,one of non-associative learning,most likely modulates the pollinating behavior patterns of empty flower visitation.Moreover,we propose that the process of habituation recovery,including spontaneous recovery and dishabituation,may also modulate the behavior of pollinators,which leads to ecological consequences of long-distance pollen dispersal and high outcross pollination rate.We believe that utilizing the methodology of non-associative learning behavioral neurobiology paradigm to investigate pollinator behavior will establish novel insights into the sensory responses and neural activity of pollination behavior in the pollination systems.

Determination of Amantadine Residue in Honey by Solid-phase Extraction and High-performance Liquid Chromatography with Pre-column Derivatization and Fluorometric Detection

Amantadine （AMA） is an anti-viral drug used in apiculture to protect honeybee against the sacbrood virus （Morator aetatulae）. This study described a reliable high-performance liquid chromatographic （HPLC） method for analyzing AMA in honey using a solid-phase extraction （SPE） cartridge （Plexa PCX） for purification, 4-fluoro-7- nitro-2,1,3-benzoxadiazole （NBD-F） as a pre-column derivatization agent, and fluorometric detection （λex =470 nm, λem=530 nm）. The chromatographic separation was performed on an XDB C18 column （150×4.6 mm i.d.） using 0.1% trifluoroacetic acid/acetonitrile （35 ; 65, V/V） as the mobile phase at a flow rate of 1.0 mLomin 1 with a run time of 20 min. Under these optimal conditions, a linear relationship was observed in the range of 0.025--1.0μg·mL-1 with a good correlation coefficient （0.998） and low limit of detection （0.0080 μg·g-1）, the recoveries were all above 90%, and the intra-day and inter-day precision （RSD） ranged from 3.4%--5.1%.