| Powdery mildew, caused by Uncinula necator (Schw.) Burr., is the most ubiquitously damaging fungal disease of grapevines worldwide. China is one of the major centers of origin of Vitis species with the potential as sources of disease resistance. In our lab, by mRNA DDRT-PCR (mRNA differential display reverse transcription-PCR ) and RACE (Rapid amplification of cDNA ends ) , the novel GLOXrg gene of Chinese wild Vitis was cloned from Vitis pseudoreticulata Baihe-35-1, which has a high resistance to Uncinula necator.In our research, the physical and chemical character and the function of GLOXrg were predicted by the usual tools and softwares of bioinformatics. GLOXrg was cloned as fusion with GST, and expressed, and the fusion protein was purified, which provide the excellent tools for detecting further its function of resistance to Uncinula necator. Fusion protein GST-GLOXrg was isolated and used to raise the polyclonal anti-(GST-GLOXrg) in rabbits. These offer powful base for the research of the specific features of GLOXrg spatial and temporal expression and the detecting of transgenic grapevine plants. The research results are as follows:1. The prediction results of physical and chemical character and function of the induced protein of GLOXrg shows that GLOXrg was probably glyoxal oxidase.which catalyzes the oxidation of a number of aldehyde and a-hydroxy carbonyl compounds, reducing O2 to H2O2 in the process.But molecular weight of the induced protein of GLOXrg was smaller than knowed glyoxal oxidase of fungi. By the analysis of the disease-resistant H2O2, GLOXrg probably play a crucial role in the resistance to Uncinula necator.2. Based on the cDNA full sequence of GLOXrg gene of Vitis pseudoreticulata, we designed a pair of specific primers, and obtained DNA band of about 1572 bp length by PCR from the first strain of 5'RACE cDNA of Vitis pseudoreticulata Baihe-35-1. The sequencing result shows that the factual length of the open reading frames is 1572 bp, and the sequence is... |
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