Optimization Of Artificial Rearing Methods For Carposina Sasakii And Expression Pattern Analyses Of Diapause Hormone Gene

Carposina sasakii Matsumura is the major pest which causes severe damage to fruits in northern China.To further strengthen the research of Carposina Sasakii,this study has confirmed the optimal combination of female and male pest numbers,the optimum temperature and humidity for female adult pests to lay eggs and hatch them,and the effect on the amount of falling eggs by different colors of egg cards,the expression pattern of diapause hormone gene was explored.It is most suitable for the adult pests to lay eggs when the relative humidity was90% with a temperature of 25°C and the number ratio of female and male pests was2+3,the number of eggs laid by single females was 153.;It was most suitable for hatching eggs when the relative humidity was 90%,and the temperature was between23 °C-31 °C,the highest hatching rate was 97.3%.In red,green,white and black,The amount of eggs laid on the white egg card was much higher than the other egg cards.The cDNA sequence of the diapause hormone gene(CsDH-PBAN)of Carposina Sasakii was cloned.The cDNA sequence length of CsDH-PBAN was 730 bp,the open reading frame(ORF)length was 570 bp,encoding 189 amino acids with a relative molecular weight of 21.75 kDa and an isoelectric point(pI)of 8.78.The amino acid sequence encoded by CsDH-PBAN is 98% similar to Plutella xylostella and 60% to Helicoverpa zea.Real-time PCR analysis showed that CsDH-PBAN was expressed in the head of the adult,the head of the first-fruited larva under long light,the head of the pupae,the head of the first-fruited diapause larva under short light,the head of the larva after 20 days of diapause and the head of the larva after 60 days of diapause;The expression of the head of the first-fruited larva under long light was the lowest,and the expression of the head of the scorpion was increased.The expression level was 3.12 times that of the head of the long-lighted fruit larvae.The highest expression was found in the head of adults,which was 15.21 times higher than that in the head of he long-lighted fruit larvae.The expression of CsDH-PBAN in the head of the first-fruited diapause larva under short light was 3.53 times higher than that in the head of the firstfruited larva under long light.The expression of CsDH-PBAN in the head the larva after 20 days of diapause decreased,which was 1.14 times higher than that in the head of the first-fruited larva under long light.At the head of the larva after 60 days of diapause,the expression of CsDH-PBAN increased significantly,which was 6.98 timeshigher than that in the head of the first-fruited larva under long light.The expression level of CsDH-PBAN was positively correlated with the activity of life and negatively correlated with diapause.The expression of CsDH-PBAN might be related to energy metabolism.