RNAi Vector Construction Of CYLV CP Gene And Sugarcane Plants Transgenic Research

Sugarcane (Saccharum officinarum L.) is a kind of important sugar and commercial crop plant in tropical and subtropical areas. The sugarcane holds for60percent of global sugar products. But the disease of sugarcane has greatly influenced the sugar content and output. Sugarcane yellow leaf syndrome (SCYLS) was detected in the1990s first in Hawaii, and was found in2001in China. SCYLS is one of the most important diseases around the word, which can cause decline in sugar quality and quantity.Sugarcane yellow leaf virus (SCYLV) is the pathogen of SCYLS. This virus belongs to Luteoviridae family, and is spread by the aphid. The genome of SCYLV is a sense ssRNA, and includes6open reading frames (ORFs). The ORFs3can code the coat protein (CP), which was proved related to many important functions of the virus.RNA interference (RNAi) is one of the fast developing techniqus in the genetic engineering. The theory of RNAi is the dsRNA can induce and control the expression of homology genes, and the genes will be silent. The specialty of RNAi is greatly efficient and specific in distinguish the genes. This techniqus is using wide in testing the genes'function, opposing the diseases and improving the breed in botany.The genetic background of sugarcane is complex and the period of common breeding is long. On the other hand, the CP kind of sugarcane is high sensitive to SYLS, which is one of the main parents of China's sugarcane. So the genetic engineering is an effective method to improving the sugarcane breeding.In this study, the CP gene of SCYLV that was isolated in Hainan was cloned, sequenced and study the relationship between SCYLV isolates. The sense stand an antisense strand of CP gene was cloned and plugged in the vector bone pCAMBIA3300. This recombination plant expressional vector i19-CP-ZF-3300included a sense stand, an intron and a antisense strand and the vector was genetic transformed into the Agrobacterium. Through the Agrobacterium-mediated transformation method, the il9-CP-ZF-3300was transformed into the callus of sugarcane. After the tissue culture and the challenge of PPT, the transgenosis plants was gained. PCR testing and RT-PCR testing was using to test the transgenosis plants, which were prepared for the last research.