MRNA Differential Display Of Laodelphax Striatellus Group Of Different Virus Transmission Ability

Rice stripe disease is caused by Rice stripe virus (RSV) which cause serious damage to rice in esatern Asia. The virus is transmitted by Laodelphax striatellus in the way of circulative,propagative transmission.Here we constructed a high affinity virus transmitter group and a low affinity virus transmitter group.DDRT-PCR(differential display RT-PCR) was exploited to achieve high efficient screen for differentially expressed genes and predicate the possible funcation of differentially expressed genes.After continual several generations' artificial selective breeding,a high affinity virus transmitter group and a low affinity virus transmitter group were constructed successfully.Incidence of RSV rised to about 90.0% while initial incidence was 16.4% in high affinity transmitter group.Incidence of the low affinity transmitter group fell to about 2.5%.Incidence of the high affinity transmitter could keep about 90.0% although without artificial selecting pressure for three generations.High affinity transmitter group is significant for the research of virus transmission mechanism, variability of pathogenicity of RSV, varieties resistance.In order to understand the molecular mechanism of RSV transmission,the differential gene expression between the high affinity virus transmitter group and the low affinity virus transmitter group was detected with the technique of DDRT-PCR.The results showed that differentially expressed bands could be amplified with four in eight random primers.Thirty-five differentially expressed bands were screened altogether,of which six were selected to identify by RNA dot hybridization.Four differential expression fragments were cloned with PMD18-T vector and sequenced.Fragment DD2 appeared in the high affinity transmitter group was similar to 5-hydroxytryptamine receptor 1D(5-HTR1D) gene.Fragment DD3 appeared in the high affinity transmitter group was similar to gyrase B gene.Fragment DD4 appeared in the low affinity transmitter group was unknown,which homologous genes were not found in GenBank by Blast analysis.Fragment DD5 appeared in the high affinity transmitter group was similar to 60S ribosomal protein L40 gene and ubiquitin gene.