Cloning And Reproductive Regulation Of TryP And PGL Genes Of Adelphocoris Suturalis

Adelphocoris suturalis(Hemiptera: Miridae)is one of the most destructive pest in Yangtze River cotton growing regions of China.It is difficult to control the A.suturalis because of its quickly activity.Understanding the development mechanisms of the reproductive regulation of A.suturalis could provide new ideas for its prevention and treatment,as well as alternative genes for the cultivation of new transgenic plants.Based on the comparative transcriptome sequencing data obtained by our research team,we found that the functions of trypsin precursor(AsTryP)and pepsinogen-like(AsPGL)genes may be related to fertility of A.suturalis.In this study,RACE(rapid-amplification of cdna ends),qPCR(Quantitative Real-time PCR)and RNAi(RNA interference)were used to verify the role of TryP gene and PGL gene in fertility of A.suturalis.The main conclusions are as follows:1.Cloning and spatiotemporal expression analysis of AsTryPThe ORF(open reading frame)region of AsTryP is 873 bp.AsTryP encods 290 amino acids with isoelectric point(pI)of 5.04 and molecular weight of 73.04 kDa.Spatiotemporal expression analysis results show that AsTryP is highly expressed in 4th instar of nymphs,followed by 3rd,5th instar of nymphs and 8-day-old females.In different tissues of 8-day-old females,AsTryP exhibited the highest levels of transcription in thorax,the second is ovary and fat body.2.The effect of AsTryP on the fertility of A.suturalisAfter the interference of AsTryP,the expression of AsTryP was significantly inhibited in the whole reproductive phase.After microinjected with dsAsTryP,the number of eggs of 11-day-old females decreased by 26.5%,the lifetime fecundity of females was reduced by 21.14%,the adult longevity of females was shortened by 2.51 days,compared with the dsGFP control.But the pre-oviposition period and egg hatching rate did not exhibit significant differences between the ds AsTryP treatment and the dsGFP control.3.Cloning and spatiotemporal expression analysis of AsPGLThe ORF region of AsPGL is 1137 bp.AsPGL encods 378 amino acids,with pI of 5.07 and molecular weight of 93.72 kDa.Spatiotemporal expression analysis results show that AsPGL exhibits the highest levels of transcription in ovary,with high levels also detected in the fat body.5.The effect of AsPGL on the fertility of A.suturalisAfter dsAsPGL injected,the expression of AsPGL was significantly inhibited in the whole reproductive phase.The number of eggs in the ovary was reduced by 23.5% and the lifetime fecundity of females was reduced by 14.18% compared with the control group,after down-regulating the expression of AsPGL.The pre-oviposition period and egg hatching rate and adult longevity did not exhibit significant differences between the dsAsPGL treatment and the ds GFP control.It indicates that AsTryP plays an important role in female fertility and longevity of A.suturalis and AsPGL also played a crucial role in reproduction of A.suturalis.It laid a foundation for understanding reproductive regulation of A.suturalis...