The Screen Of The Differentially Expressed Genes Of Flight Muscle Cell Apoptosis In Macrosiphum Avenae Fabricius After Migration

Our previous work found that the muscle cells do apoptosis after Macrosiphum avenae long distance migration. This study was performed to screen the differential expressed genes during the muscle cell apoptosis, with the molecular techniques of RT-PCR, DDRT-PCR, QPCR and RACE. The results of our work help to know the molecular mechanism of aphid migration, from which we can find new way for decreasing the disaster. The results and conclusions as follows,1. The ubiquitin-ribosomal S27a gene's encode region (GenBank: HQ285247) of Macrosiphum avenae was cloned using RT-PCR and RACE. Full length is 507 bp, translation area is 453bp for 150 amino acids. The results of homologue contrast to other insects shows that, ubiquitin-ribosomal S27a gene has high conservation, 77.92%.2. Also, the ubiquition gene encode region (GenBank: HQ285248) of Schizaphis Graminum (Rondani), was cloned. The heredity of the three aphids plus Acyrthosiphon pisum (Harris) has been determined from gene ubiquitin and confirmed the model of protein histysis.3. With the technique of DDRT-PCR, the differential expressed genes after the long distance migration of Macrosiphum avenae were screened. In this study, almost all of the mRNA of Macrosiphum avenae had been cloned out, using the 3rd DDRT-PCR primers: 3 anchor primers and 8 random primers, totally 24 combinations. Through the 6% polyacrylamide gel electrophoresis, 64 differential expressed mRNA bands has been screened, including 8 specific expressed and 17 non-expressed, 16 up-regulated and 23 down-regulated gene bands after migration, named P53-like regulator of apoptosis and cell cycle, Ras-related protein Rab-39B-like, Beta-galactosidase, Glucose transporter 1, Probable maltase L-like et al. Besides we also compared the effects in separating the differential expressed genes between the 6% polyacrylamide gel electrophoresis and improved high concentration agar gel electrophoresis. The latter screened out more than 70% of the former differential expressed genes, especially the specific expressed genes. So we can say that the latter can play an important role in screening the specific expressed genes in experiments like inducing by single intense mannully treatment.4. The expression of Macrosiphum avenae ubiquitin-ribosomal S27a gene of each instar and its different parts was analysised by QPCR. The result shows: the expression of ubiquitin-ribosomal S27a gene in abdomen is far higher than in head and thorax. In the 5th day, the first day after migration, the expression in all three parts show an obvious increase and then decrease to previous. It dedicates that the ubiquitin-ribosomal S27a gene plays unreplaceable role in proteins histysis and translocation of the energy in Macrosiphum avenae, stimulus the behaviors change from migration to breeds. And also the way to understand the molecular mechanism of migration and the muscle cell apoptosis was made possible.