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根据Banerjees等（1992）的方法，建立了中国小麦花叶病毒(CWMV) 和大麦黄花叶病毒(BaYMV)外壳蛋白（CP）叶绿体离体跨膜运输体系，研究了孵育时间、CP浓度对跨膜运输效率的影响。结果表明，CWMV CP和BaYMV CP可分别快速进入离体的小麦与大麦叶绿体中，其跨膜运输所需的孵育时间均最低为5 min，孵育时间超过15 min后进入叶绿体中CP的量不受影响；加入跨膜体系中CP的浓度与跨膜后进入叶绿体的CP浓度呈正相关，能够实现跨膜的最低CWMV CP和BaYMV CP浓度分别为4.2和37.8 μg·mL^-1。

英文摘要:

Referred to Banerjee's method, the systems of importing the coat proteins of Chinese wheat mosaic virus and Barley yellow mosaic virus (CWMV CP and BaYMV CP) into intact chloroplasts in vitro were established, the effects of incubation duration, the concentration of CWMV CP and BaYMV CP on import efficiency were studied. The results showed that both CWMV CP and BaYMV CP was able to rapidly import into isolated chloroplasts of wheat and barley. In the test, the lowest incubation duration of CWMV CP and BaYMV CP importing into the chloroplasts were both 5 min, the amounts of CWMV CP and BaYMV CP imported into the chloroplasts were not increased with the extension of incubation time after 15 min. Besides, the relation between the amounts of CWMV CP and BaYMV CP imported into the chloroplasts and the amounts of CWMV CP and BaYMV CP added in the import system was significantly positive, the lowest concentrations of CWMV CP and BaYMV CP in the import system was 4.2 and 37.8 μg·mL^-1, respectively. The establishment of CWMV CP and BaYMV CP importing into intact chloroplasts in vitro could supply foundation for studying the pathogenesis of CWMV and BaYMV.

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