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采用花椰菜抗黑腐病近等基因系C731(感病系)和C712(抗病系)作为材料,利用cDNA-AFLP技术,研究了花椰菜黑腐病抗性在黑腐病菌侵染和非侵染条件下基因表达的情况.获得了两个阳性克隆M2和M6.Northern杂交和点杂交进一步证明M2是组成型表达的cDNA片段,只是在病菌侵染与非侵染条件下表达丰度不同.而M6是差异表达的cDNA片段.Southern杂交表明M6 cDNA片段在花椰菜基因组中是单拷贝序列.随后的序列分析发现M6 cDNA片段与拟南芥1号染色体的BAC F19P19中66665~66813bp有84%同源性,该片段编码拟南芥的2A6蛋白的部分序列.推测的氨基酸序列与拟南芥的2A6蛋白部分序列有91%同源性.用H2O2作为外源分子胁迫处理的初步功能分析发现:M6 cDNA片段受H2O2诱导,在诱导早期16~24h高度表达,其在叶片中的积累呈逐渐上升趋势.根据序列分析和初步的功能分析的结果推测:M6 cDNA片段可能就是编码花椰菜中类2A6蛋白的部分基因片段.是参与花椰菜抗病反应信号途径的相关调控基因片段.
Using cauliflower resistant to black rot disease near isogenic lines C731 (susceptible line) and C712 (resistant line) as materials, the effects of black rot resistance of cauliflower on the infection and non-infection of black rot virus were studied by cDNA-AFLP technique. Under the conditions of gene expression obtained two positive clones M2 and M6.Northern hybridization and dot hybridization further confirmed that M2 is constitutively expressed cDNA fragments, only in the infection of bacteria and non-invasive conditions, the abundance of different expression M6 is a differentially expressed cDNA fragment.Southern hybridization shows that the M6 cDNA fragment is a single copy sequence in the genome of cauliflower.Analysis of the subsequent sequence found that the M6 cDNA fragment has 84% homology with 66665 to 66813 bp in BAC F19P19 of Arabidopsis chromosome 1 This fragment encodes a partial sequence of Arabidopsis 2A6 protein.The deduced amino acid sequence is 91% homologous to part of the 2A6 protein sequence of Arabidopsis thaliana Preliminary functional analysis of H2O2 as an exogenous molecular stress treatment found that M6 The cDNA fragments were highly induced by H2O2 and highly expressed in leaves at 16 ~ 24 h after induction, and the accumulation in leaves increased gradually.According to the results of sequence analysis and preliminary functional analysis, it was presumed that the M6 cDNA fragment may encode the Cauliflower Portion 2A6 protein gene fragments. Regulatory gene fragment is involved in the signal pathway resistance Cauliflower reaction.