Study On The Expression Of Some Cold Resistance Genes And Some Amino Acids Of Overwintering Euzophera Pyriella

Euzophera pyriella is one of the most important pests in the forest and fruit industry in Xinjiang,it overwinters with mature larvae,it at the place where the trees become warped or is harmful.In order to explore the diapause characteristics of larvae of Euzophera pyriella during diapause period,the cold resistance related genes-the Heat shock protein gene,Fatty acid desaturase gene,and other genes related to diapause,such as arginine kinase,acetyl-CoA carboxylase,Fatty acid synthase,Glycogen phosphorylase,hexokinase,and amino acid were selected as research objects to study their expression or change rule,aiming at promoting the exploration of diapause mechanism of Euzophera pyriella,and providing new ideas for integrated Euzophera pyriella control.(1)Screening of reference genes for diapause period and low temperature stress of Euzophera pyriella:under 4? low temperature stress,the stability of the reference gene of Euzophera pyriella is UBQ>GAPDH>Actin>18s>TUB;Under different low temperature stress for 1h,the stability of the reference gene is UBQ>18s>GAPDH>Actin>TUB.The stability of reference genes in overwintering diapause of Euzophera pyriella is GAPDH>18s>UBQ>TUB>Actin.UBQ can be selected as the reference gene under short-term temperature stress of Euzophera pyriella,and GAPDH can also be used as an reference gene in diapause research.(2)Expression characteristics of Heat shock protein(HSP60,HSP70,HSP90)genes: The expression of HSPs gene was significantly up-regulated after 4? low temperature stress,reaching the peak at 1 hour,and began to decrease at 3h,and then increased again.The differential expression of HSP60 was significantly lower than that of HSP70 and HSP90 genes.Under different low temperature stress for 1h,there was no significant difference in HSPs gene expression under 10? low temperature stress.The expression increased significantly under 5? stress and began to decrease under 0?.The expression level of HSP60 and HSP70 genes increased under lower temperature stress,while the expression level of HSP90 genes remained at a low level.All three genes reached their maximum at-15?.The expression trends of the three heat shock protein genes during overwintering diapause are similar.They began to increase significantly in September,and began to decrease and increase again in October.The three genes were at high expression levels in the middle and late stages of diapause and continued until the end of diapause.(3)The expression of fatty acid desaturase(FADs)gene was up-regulated during diapause,FAD-?6increased first and then decreased during diapause,reaching the highest peak in January.FAD-?9 was highly expressed before diapause,and it rose first during diapause,then declined after December,but remained at a high level;The expression trend of FAD-?11 is similar to FAD-?9,but the highest peak appears in November.The expression of ACCase gene was relatively high during diapause,and it began to increase in September,then decreased first and then increased;AK gene continued to be highly expressed during diapause and did not decrease.The expression of FAS gene reached the maximum in November,but in September and October it was also significantly higher than that in June,the expression level reached the June level rapidly in December;GP gene has the same trend as AK gene;HK gene was the lowest when it was about to enter diapause,after entering diapause,it began to increase gradually,similar to the expression abundance in June,and began to decrease at the later stage of diapause.(4)Changes in total amino acids and amino acid composition of larvae of Euzophera pyriella in diapause period: The total amount of 16 amino acids in each period is not significantly different.Threonine,arginine,alanine,valine,methionine and lysine increased significantly with the increase of diapause time.While glutamic acid,histidine,glycine,tyrosine and cystine decreased significantly.The changes of alanineand cystine are the most significant in this process,it may be related to diapause metabolism.In conclusion,UBQ or GAPDH were selected as the internal reference genes according to the short-term low temperature stress or diapause period,HSPs?FADs?ACCase?AK?FAS?GP and HK genes fluctuate a lot in diapause period of Euzophera pyriella,except FAD-?6 gene and HK gene,other genes were highly expressed in September before diapause,these genes are closely related to the diapause of Euzophera pyriella,there are great differences in other periods,which are closely related to the metabolism of insects,the changes of amino acid contents in diapause stage also confirmed the complexity of metabolism,the results provide data for further study on diapause mechanism of Euzophera pyriella.