Effects Of Temperaure On The Expression Of Metabolic Enzymes And Stored Protein Gene Of Soybean Pod Borer In Diapause

Soybean pod borer Legumonivora glycinivorella Matsumura is an important pest on soybeans.It is a specialized diapause pest that is harmed by earthworms.It damages the quality and yield of soybeans because it is eaten.It overwintered with mature larvae,and the diapause period was about 10 months,making it difficult to control.It is clear that the metabolic changes of the soybean diapause-diapause-diapause-relieving process can provide us with a scientific basis for accurately measuring the occurrence period of soybean moths,and then doing a good job of prevention and control to reduce the use of pesticides.In this paper,the soybean borer Leguminirora glycinioorella treated at 4℃,13℃,18℃,23℃,25℃and outdoor natural conditions at different temperatures in not diapause(before diapause,September)-diapause(diapause start,November;mid-diapause,February)-diapause termination(June).The changes of the activitity of the main metabolic enzymes(trehalase,THL;sorbitol dehydrogenase;SH,pyruvate kinase,PK;peroxidase,POD)for anti-freeze and the expression of storage protein hexamerin at different stages to provide the basis for the future occurrence of soybean borer in soybean.In this study,kits were used to determine the changes in the expression levels of the above four enzymes in soybean borers before and after diapause,after treatment at the induced diapause temperature and at various stages of diapause.Data analysis was performed using SPSS 23.0 and Excel data analysis software.Here are the main results:(1)Under natural conditions,the activities of THL,SH,PK,and POD in soybean body from pre-diapause(September)to early diapause(November)were significantly changed.THL activity decreased from 11.37 U/mg to 10.34 U/mg,SH activity decreased from 3.84 U/mg to 0.98 U/mg,and PK activity decreased from 9.10 U/mg to 1.23 U/mg,however,PK activity increased from 6.53 U/mg to 8.12 U/mg.(2)Soybean borer pods were treated with different induction temperatures at 4℃,13℃,18℃,23℃,and 25℃ until the early diapause(November).The activities of the four enzymes PK,THL,SH,and POD were significantly changed,and they were significantly different from those of the control group under natural conditions.Under 4℃ induction,the activity of THL,SH,and POD in soybean was lower than that of natural.The activities of these three enzymes were significantly reduced,which was decreased by 4.41 U/mg,2.88 U/mg,and 4.51 U/mg,respectively,while the enzyme activity of PK was significantly increased by 5.55 U/mg.(3)Soybean borer treated at different induction temperatures at 4℃,13℃,18℃,23℃,and 25℃ until the mid-diapause(February).There were significant differences in the activity of the 4 kinds of metabolic enzymes PK,THL,SH,and POD.The diapauses of diapause soybean fruit moths treated under different temperatures at 4℃,13℃,18℃,23℃,and 25℃ can be eliminated under the outdoor conditions,and the dip between the 23℃ and 25℃ soybean dregs and the natural conditions can be eliminated.Worm easier to relieve diapause.In mid-diapause(February),under 4 ℃ induced soybean pod borer,(1)SH activity was significantly higher than the 25℃ induction treatment;(2)The activity of PK and THL was similar to that of the enzyme-inducing activity of soybean borer in 18℃;(3)The activity of PK in soybean pod borer was lower than that in 18℃;(4)During the mid-diapause period,the activities of THL,SH and POD in soybean pod borer treated with 18°C were higher than those in the other 4 temperature-treated soybean pod borer;(5)The above 4 kinds of metabolic enzyme activities in soybean pod borer diapause mid-period caused by 25℃ were similar to the corresponding 4 kinds of enzyme activities under natural conditions.During the period of diapause withdrawal(June),the PK activity of soybean pod borer induced by 4 ℃,13 ℃,18 ℃,23 ℃,and 25 ℃ was higher than that of the above five temperature-treated soybean borers in the early diapause and diapause mid-stages,but thePK activity in the above five temperature-treated soybean borer did not differ significantly during the diapause-relieving period.(4)The analyze to the change of metabolic enzyme activity during the whole diapause period indicated that the activity of THL、SH and POD in soybean pod borer induced to diapause by five diapause-inducing temperature(4℃、13℃、18℃、23℃and 25℃)reached maximum value in February as 15.25 U/mg、1.94 U/mg and 35.56 U/mg respectively,and then gradually decreased,while the activity of PK reached minimum value at the same time as 1.57 U/mg and then gradually increased.So,February is the turning point of metabolic process in soybean pod borer during diapause period,it’s a critical period to change from deep diapause to diapause termination,known as awaking stage,meanwhile,the metabolism change from weak to strong.(5)By induced with different temperature,the expression of storage protein Hexamerin in diapausing soybean pod borer varied significantly among different diapause stages.The expression of the protein hexamerrin in the 4℃ induced soybean pod borer was significantly higher than that in the natural condition,and the difference was significant.Soybean pod borer reached its peak at the beginning of diapause(November),diapause transition period(February)and diapause release(June),5.58 U/mg,9.84 U/mg,and 10.11 U/mg,respectively.In February,there was a turning point in its expression.