| Brood pheromones is an important chemical communication substance.Its main components are 10 kinds of esters.In recent years,researchers have discovered an important brood pheromone component,E-beta-ocimene.It can not only inhibit the bees ovarian mature,also involved in the regulations of the worker foraging behavior,increased the proportion of workers,and postponed the first collecting date of foragers.However,the determination of E-beta-ocimene content in larvae surface,and the differences in the expression of E-beta-ocimene related genes between ages are not sufficiently studied.In this study,GC-MS/MS was used to determine the content of E-beta-ocimene on the body surface of worker larvae of 2-6 days of age.It revealed that the larvae of workers at different days of age were different in the content of the body surface of E-beta-ocimene.QPCR technology was used to explore the differences in expression of related genes in different day old larvae.The results are as follows:In the first part,the content of E-beta-ocimene.in the epidermis of honeybee larvae was determined by gas chromatography-mass spectrometry(GC-MS/MS).The samples of 2 to 6 day old larvae of workers in Apis mellifera were collected and extracted with n-hexane solution and preserved at-20 ?.GC-MS/MS collected data and compared with Nist library,We have a qualitative and quantitative analysis of E-beta-ocimene.The resp.Lts showed that the content of E-beta-ocimene in bee larvae changed with the age of bee larvae.The content increased with the increase of age,reaching the maximum value at 4 days of age,about 2.225?g,but the body surface content of 5 and 6 days larvae gradually decreased.The difference significance analysis showed that there were significant differences in the content of E-beta-ocimene,but there was no significant difference between 5 and 6 days larvae.The weight of the larvae increased exponentially with the growth of the larval age,and the weight difference between the larvae of 2 to 6 ages was very significant.Compared with the weight of the corresponding days of age,the relative content of E-beta-ocimene on the surface of the larvae of 2,3 and 4 days is higher,and the relative content of 5 and 6 days is lower,that is,the content was relatively high in the small larva stage,while the content in the large larval stage was relatively low.The second part,real-time PCR(qPCR)was employed to determine and analysis the expression of related genes about E-beta-ocimene in 2 to 6 days old in Apis mellifera.The res?Lts showed that the relative expression of 3-har,mk-like,pk-like and fps was not significant.The relative expression level of aatc-like,md and llp-like was higher at 2 days old,especially the gene aatc-like.It has high expression at the age of 2 and 3 days,which is 2 to 3 times of the expression of 5 day age.The expression level of hs 1 was not high at any age of day,but the expression level of gene decreased gradually with the increase of age.Interestingly,the relative expression of gene aatc-like,md,llp-like and at 1 was up at the age of 6 days,and it was close to the relative expression of 2 days old.The up-reg?Lated expression of these genes at the age of 6 days may be related to the time that the larvae are entering the capping period. |
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