Study On Import Of Plant Virus Coat Protein Into Intact Chloroplasts Through Membrane In Vitro

Tobacco mosaic virus(TMV), a typical member of Tobamovirus, which hold huge host extent. Tobacco mosaic disease infected by TMV was the leading disease in crop region of tobacco all over the world continuously. Rice stripe virus(RSV), a typical member of Tenuivirus, was an important rice virus in 16 province or city in China. It had infiicated damage over a huge hectarage from 2000 in Jiangsu province and its neighber areas.Two antiserum against RSV, Sv11 or Sv21, and two antiserum against TMV, Mv31 or Mv41 were produced with the purified virus, the diluted times of the four antiserum were 1:65536, 1:16384, 1:2048 and 1:8192. We could detect virus in infected plants by Sv11, Sv21 and Mv41 with high detection accuracy for 1:16384, 1:8192 and 1:12800 by indirect ELISA successfully, but the three antiserum were less sensitive to health plants.Effects of time and the amount of TMV-CP on import efficiency and species selection of import of TMV-CP into chloroplasts were studied. So we could prove up that the characteristic of importing TMV-CP into intact chloroplasts in vitro and its species speciality. Results showed TMV-CP was rapidly imported into isolated chloroplasts of tobacco. And the amount of TMV-CP imported into the chloroplasts was independent of the import time, if the time preponderate over 15 min. Besides, the relation between the amount of TMV-CP imported into the chloroplasts and the amount of TMV-CP added in the import was significantly positive correlation. Furthermore, our studies showed that TMV-CP possess species selection when TMV-CP crossed the membrane of chloroplasts, this was why TMV can not infect gramineous plants.The system of import of Rice Stripe Virus Coat Protein(RSV-CP) into chloroplasts in vitro was established, effects of time and the amount of RSV-CP on import efficiency and species selection of import of RSV-CP into chloroplasts were studied first. Results showed RSV-CP was rapidly imported into isolated chloroplasts of rice and wheat. And the amount of RSV-CP imported into the chloroplasts was independent of the import time, if the time preponderate over 15 min. Besides, the relation between the amount of RSV-CP imported into the chloroplasts and the amount of RSV-CP added in the import was significantly positive correlation. Furthermore, our studies showed that RSV-CP possess species selection when RSV-CP crossed the membrane of chloroplasts, this was why RSV can not infect nonhost plants such as nightshade family of plants.We found infection of import time and the amount of CP to import efficiency for RSV-CP and TMV-CP were the same. The amount of CP imported into the chloroplasts was independent of the import time, if the time preponderate over 15 min. Besides, the relation between the amount of CP imported into the chloroplasts and the amount of CP added in the import was significantly positive correlation. Furthermore, studies showed that two virus-CP possess species selection when CP crossed the membrane of chloroplasts.