Study On Chiral Configuration Of Honeybee JH And The Effect Of JHA On The Titer Of Honeybee JH

Honeybee is a typical social insect,with complete population biological characteristics,different morphological characteristics and clear division of labor within the population,forming a closely interdependent complete population.Juvenile hormone(JH)is a kind of sesquiterpenoids synthesized and secreted by the corpora allata(CA)cells of insects.It is mainly used to regulate the growth characteristics of larvae,promote ovarian maturation and prevent the larvae from entering the next instar early.JH is also involved in a series of important physiological and biochemical processes,such as body appearance,molting,diapause,stress resistance and stimulation of yolk formation in female adults.At present,the analysis method of JH is mainly to determine its titer,and does not involve the chiral configuration analysis of JH enantiomers.In order to explore the enantiomeric configuration of honeybee JH,a chiral enantiomeric analysis method of honeybee JH was established;The effect of JHA on JH titer of honeybee larvae was studied.1.Study the separation of JH under the conditions of different packing chiral column chromatography,mobile phase and instrument.The results showed that: under the optimized conditions,10 chiral peaks of JHⅠ,JHⅡ and JHⅢ could be separated by ultra performance liquid chromatography-mass spectrometry(UPLC-MS/MS)with a reversed-phase coated chiral column of polysaccharide derivatives and a mobile phase of 1% formic acid water acetonitrile.The results showed that the coated chiral column with amylose tris(5-chloro-2-methylphenylcarbamate)coated on silica gel had a good separation effect for JH.The results showed that the detection methods of JHⅠ,JHⅡ and JHⅢ had good sensitivity and reproducibility,and met the detection and analysis standards.2.The enantiomers of R-JHⅢ and S-JHⅢ were successfully prepared by liquid chromatography.The time-dependent density universal inclusion theory(TDDFT)was used to construct each enantiomer and predict the ECD.The absolute configuration of the two enantiomers of JHⅢ was determined and the optical rotation of each enantiomer was detected.Compared with the mass spectra of honeybee larvae,the results showed that only S-JHⅢ enantiomers were synthesized in honeybee.This is different from the R-JH in most other insects,which indicates that the chiral configuration of S-JHⅢ may play an important role in regulating some specific physiological behaviors of honeybee.3.To study the effect of JHA on JH titer of honeybee.Three kinds of JHA,fenoxycarb,methoprene and pyriproxyfen,were selected to treat honeybee larvae according to OECD standard.The results showed that the JH titer of honeybee larvae decreased significantly under JHA stress.In different concentrations of JHA treatment group,the JH titer of larvae was 13 μg The JH titer of 100 mg/L and 150 mg/L treatment groups was 3 μg High concentration of JHA could significantly reduce the JH titer of larvae;The JH titers of larvae were lower than 8 μg/L.These results suggest that JHA of three different concentrations can significantly reduce the JH titer of honeybee,which may be related to the different effects of JHA on the synthesis and metabolism of endogenous JH by lateral pharyngeal body.4.Using the next generation high-throughput sequencing technology,we compared the transcriptome level changes between the control group and the treatment group.The results showed that 34,532 and 222 DEGs were identified in three different JHA treatment groups and the control group.The expression of the group treated with propyl ether was up regulated by 21 and down regulated 13;Compared with the control group,222 expression was up regulated and 310 were down regulated in phenoxycarb group;Compared with the control group,155 up regulated and 67 down regulated expression in the treatment group.Two DEGs were screened which responded to three different JHA.Among them,JHE is a specific enzyme to degrade JH and regulate JH titer to hydrolyze JH.The expression of the group treated with eipronate and propyl ether increased,which reduced the p H of JH and affected the larva development.It was suggested that JHA affected the p H of bee by affecting the enzyme and synthetase of JH.The results showed that JHA could only selectively synthesize S-Body JHⅢ in honeybee for the first time,and the titer of JHⅢ in honeybee larvae decreased significantly after oral administration of low concentration JHA,which affected the growth and development of honeybee larvae.