| Sodium pyrosulfite and Glycine are two common food additives in life.Sodium pyrosulfite can be widely used as preservative and antioxidant in food industry,such as the production of preserves,biscuits and candies.Glycine is used as food flavoring agent and sweetening agent.The studies have shown that cytotoxicity and genotoxicity are involed in the Sodium pyrosulfite.Not only it can cause Chromosome aberration(CA)and Sister chromatid exchange(SCE)in human blood lymphocyte,but also has certain damage to the nervous system.The enhance level of lipid peroxidation in the kidney and liver of mices were caused by Sodium pyrosulfite.Now many researchers focus on the toxicology of Sodium pyrosulfite especially at the liver,kidney and neuron of mices.However,the influence of about Sodium pyrosulfite for Polyrhachis vicina Roger has not been reported.Glycine is one of the main elements of the central nervous system that mediating rapid inhibitory nerve conduction.Glycine has a damaging effect on excitatory nerves.Meanwhile,many dissertations have reported that Glycine solution can not only reduce the climbing ability of insect,but also influence the duration and frequency of behavior pattern,such as flying,walking,grooming,and other activities.Consequently,Glycine is potentially toxic to insect.The impact on about Glycine for Polyrhachis vicina Roger has not been reported.The estrogen-related receptor(ERR)belongs to the third subfamily of the nuclear receptor superfamily,so far,no natural ligand has been found.Therefore,as an orphan nuclear receptor,ERR structure is very similar to estrogen receptor(ER),with homology.It has been shown that ERR and ER jointly regulating various life activities in mammals.At the same time,it play an essential role in the growth and development of insect and coordination of their normal metabolism.Ecdysteroid receptor(EcR)belongs to the first subtribe nuclear receptor superfamily member,it ligand is Ecdysteroid hormones.It is the only known nuclear receptor of ligand in insect.During the growth and development process of insect,Ecdysteroid has strictly regulation on a series of important life activities of insect,such as molting and metamorphosis.The dimer composed of Ecdysteroid receptor(EcR)and UItraspiracle(USP)is the target of Ecdysteroid,through the jointing a series of transcription factors,regulating life activities of insect,such as,the growth,development and reproduction and so on.Glutamine synthetase(Gins)is a kind of enzyme widely existing in most cells of organism.Not only can it specifically catalyze glutamic acid to glutamine,but it is also a key enzyme in glutamate metabolism.Glutamine synthetase mainly involved in life activities such as regulating cell growth,energy metabolism,protein and nucleic acid synthesis,and immune response.This enzyme is also involved in maintaining normal nerve transmission in the body and repairing damaged nerves.The mutation of this enzyme can lead to the occurrence of a variety of neurocognitive diseases and also play an indispensable role in insect growth and development.Polyhachis vicina Roger is a typical social insect.One of the main characteristics is that they has obvious grade differentiation.As a complete metamorphosed insect,P.vicina are divided into four development stages including egg,larva,pupa,adult,the adult can be segmented into worker,male,and female,according to their external appearance,living habit,and social division of labor.P.vicina is also the only edible and medicinal ant designated by Ministry of Health of the People's Republic of China,it has many merits such as short development cycle and easy feeding.It is a good material for study the growth and development of social insect and the regulation of gene expression.In this experiment,the influences of Sodium pyrosulfite and Glycine were evaluated to on the appearance change,body weight and mortality of P.vicina different castes adult.Real-time fluorescence quantitative PCR(qRT-PCR)technology wao used to detecting the change in the expression of PvEcR,PvERR,PvCREB,PvGins and PvUSP at the mRNA level after treatment with Sodium pyrosulfite and Glycine,it is speculated that these two additives may have an impact on the development of the adult.of P.vicina.In order to verify the interference efficiency of food additives acting on the different castes adult development of P.vicina,RNA interference(RNAi)technology was used to acting on PvERR and PvGins genes of the adult,respectively.Then real-time fluorescence quantitative PCR(qRT-PCR)technologywas used to detecting the mRNA expression of PvEcR,PvERR and PvGins in different castes adult,statistic changes of the shape,body weight and mortality were made,and compared with the interference results of the previous two food additives,it was confirmed that these two food additives were harmful to adult of P.vicina.Furthermore,the relationship among PvEcR,PvERR,and PvGins was further analyzed from mRNA level,and the effect of PvEcR,PvERR and PvGins on the development of P.vicina were speculated.The research results are as follows:1.The adult of different castes of P.vicina were feeding by the Sodium pyrosulfite solution.Compared with the control group,through analyzing the weight of different grades,the body weight of different castes in the treated group decreased,but there was no significant difference.Compared with the control group,the survival rate of males in the treated group was the lowest,followed by workers and females.Compared with the control group,the males of the treated group appeared broken wings and remnant wings,however,the workers and females were not obvious change in appearance.Compared with the control group,the results of qRT-PCR shown that the expression level of PvEcR,PvERR,PvCREB,PvGins and PvUSP in the treated group were up-regulation and down-regulation fluctuation.From these results,it is speculated that the effects of Sodium pyrosulfite on body weight,mortality and appearance may be related to the interference of the normal expression of related genes in different grades of adult.2.The adult of different castes of P.vicina were treated by the Glycine solution.Compared with the control group,in the treatment group the body weight loss and survival rate of reduced.In the treatment group,except for the remnant wings of male,the appearance of adult insect other grades changed little.Compared with the control group,the results of qRT-PCR shown that the expression level of PvEcR,PvERR,PvCREB,PvGins and PvUSP in the treated group were up-regulation and down-regulation fluctuation.From these results,it is speculated that the effects of Glycine on body weight,mortality and appearance may be related to the interference of the normal expression of related genes in different grades of adult.3.Different castes adult of P.vicina were treated by PvERR-dsRNA solution.Compared with the control group,PvERR was down-regulation to different levels in different castes,which indicate that PvERR was successfully interfered.After PvERR was interfered,the weight of different castes adult loss and the survival rate of the different castes adult were decreased.It was found that male had broken wings,and the female shown different degrees of expansion in their abdomen,this show that the interference of PvERR has a certain effect on the body weight,mortality and appearance of adult of different grades.After PvERR was interfered,the relative expression of PvERR and PvEcR in worker,female and male were significantly decreased,and the relative expression of PvGins were up and down fluctuation in the worker and female,while it was significantly down-regulation in male.This indicate that PvERR is closely correlated with PvEcR and PvGins.This association may have adverse effect on the development of adult of different grades.After PvERR was interfered,the effects of PvERR on the body weight,mortality,appearance and related gene expression level of adult of different grades were similar to the influence of Sodium pyrosulfite and Glycine on different grades,this show that the influence of food additives on the development of adult of P.vicina may be related to the intervention of related genes in the ants.4.The workers and females were treated by PvGins-dsRNA solution.Compared with the control group,the expression of PvGins in both workers and females were down-regulation,which indicated that PvGins was the successfully interfered.Under the influence of PvGins gene interfered,the weight of workers and females loss,the mortality rate of increased.The abdomen of workers were wrinkled,while females were darkened and enlarged.After PvGins were interfered,the expression level of PvEcR and PvERR in workers and females were down-regulation.From this point of view,the change of PvGins expression level lead to the expression change of PvEcR and PvERR genes,which also indicate that there are correlation among PvGins,PvEcR and PvERR genes.This association may influence the development of workers and females.After PvGins is interfered,the effects of PvGins on body weight,mortality,appearance and related gene expression level of workers and females were similar to the influences of the two food additives on workers and females.This show that the influence of food additives on the development of adult of P.vicina may be related to the intervention of related genes in the ants. |
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