Multiple RT-PCR Detection Of Quarantine Viruses In Grape And Expression Of The Coat Protein Gene Of Arabis Mosaic Nepo Virus And Preparation Of Virus-Specific Antiserum

In China, only 9 grape and virod species was exactly identified. However, the grape and virod species were 55 in total in the world. Therefore, some plant virus diseases are quarantine plant disease to China and it is important to study the method for detecting the plant virus, which may be very useful for the development of the Chinese grape industry. By now, methods used to detect ArMV are biological detection, serological detection , imaging the virus particles and molecular biology detection in China, among which serological detection and molecular biology detection are in common use.Arabis mosaic nepo virous(ArMV)has been defined as an important quarantine plant virus(on the A2 list). By now, methods used to detect ArMV are serological detection, which need to prepare quantity of antiserum with high pureness and specificity for detecting the plant virus by serological method. But it is difficult to obtain quantity of antigen with high pureness with the traditional method of purifying the virus particles from the infected plant. And the antiserum prepared by traditional method is not absolute specificity, which may cause false positive results in the process of detecting the plant virus. In this study, we detected ArMV in the in the grape seeding imported from the other country by serological detection, and established a RT-PCR method to detect ArMV. At the same time, in order to overcome the limitation of the preparation of antiserum by traditional method, we use molecular biology method to clone the partial coat protein gene of ArMV, construct expression vector, express the coat protein and prepare the antiserum with high specificity.Multiple RT-PCR, based on RT-PCR, could offer much more information in one reaction system than the common RT-PCR. Compared with single RT-PCR, multiple RT-PCR is more quick, more efficient and more economic in detecting two and above two species virus at the same time. So it is very useful in detecting viruses and breeding the grape seed without viruses.Tomato ringsopt virus/nepovirus(ToRSV)and Tobacco ringspot virus(TRSV), which seriously infect grape, are on the A1 and A2 list of quarantine plant virus. Because they always infect grape together, it took more time and cost more money to detect them at the...