Constructing And Transforming Of Rnai Vectors For 2 Osvdac Genes In Rice

Voltage-dependent anion channel (VDAC) exist widely in fungi, plants and animals. It play an important role both in the material transport and regulation of energy metablism between mitochondria and cytoplasm, and in the process of mitochondria-mediated apoptosis. In rice genome, there are at least 8 vdac genes identified. In this study, two RNA interference (RNAi) expression vectors of two osvdac genes, osvdac3 and osvdac5, were constructed by method of Gateway recombination cloning technology, and transformed into two varieties of YTB and Nipponbare to explore their physiological function in rice. The main results are as follow:1. Four RNAi expression vectors for osvdac3 and osvdac5, named osvdac3-1, osvdac3-2, osvdac5-1 and osvdac5-2, were successfully constructed.2. Four RNAi expression vectors were transformed into YTB independently. The positive transgenic rice plants and seeds were obtained only in the group of callus transformed by RNAi expression vector osvdac3-2. Statistical data showed that the interfered YTB callus has a lower differentiation rate. And transgenic plants has a lower positive rate and seeds-setting rate than the control of wild type YTB.3. The resistant callus of Nipponbare transformed with the RNAi expression vectors, osvdac5-1 and osvdac5-2, independently or together can not differentiate into plantlet in the differential medium. Therefore, RNA extracted directly from resistant callus and wild type Nipponbare callus was used for real-time PCR to analyze the quantity of osvdac5 expression. The result showed that the RNA expression level osvdac5 in resistant callus infected by RNAi vector osvdac5-1 is much lower than that of wild type callus. It is demenstrated that the interfering gene osvdac5 in resistant callus was successfully carried, but why there was no differentiation of resistant callus is still to be verified.