Rao Zhi-Ming, Dong Hai-Tao, Zhuang Jie-Yun, Chai Rong-Yao, Fan Ye-Yang, Li De-Bao, Zheng Kang-Le
National Center for Rice Improvement, China National Rice Research Institute, Hangzhou 310006, China.
Yi Chuan Xue Bao. 2002 Oct;29(10):887-93.
A pair of near isogenic lines G205 and G71 were selected from recombinant inbred lines (RIL) of Zhong156 x Gumei2. On the resistance locus Pi-25(t), G205 had the resistant allele that was from Gumei 2 while G71 had the susceptible allele that was from Zhong156. For the genetic background, different alleles were detected on only 24 loci out of the 672 RFLP or SSLP loci surveyed. The expression profiles of G205 and G71 in response to Magnaporthe grisea were investigated using cDNA microarray containing 2200 Expression Sequence Tags (ESTs). The leaves were inoculated with the pathogen for 12 hours at 4-leaf stage and 998 genes were identified in total. Three genes were up-regulated significantly by the fungus in G205 only. The functions of two genes were known but that of the third gene were unknown. The two genes encoded casein kinase II alpha subunit and retrotransponson TOS17 insertion element respectively. Other thirty-five genes had similar expression patterns between NILs. Among them, 17 genes were up-regulated while 18 genes were down-regulated by the inoculation. The functions of 33 out of the 35 genes were known. BLAST analysis showed that all thirty-five. BLAST analysis showed that all thirty-five genes with known functions were relative to defense reactions, signal transduction, stress response, photosynthesis and sugar metabolism. Northern blot confirmed that four of five differentially displayed genes randomly selected had the same expression patterns as those detected in cDNA microarray. Two of them were up-regulated genes encoding casein kinase II alpha subunit and glycine-rich protein (Grp), and the other two down-regulated genes encoding nitrilase-associated protein and 18S small subnit ribosomal RNA gene respectively. Northern blot also revealed that the expression of Grp was consistently up-regulated from 0 to 36 h after the inoculation of the fungus. These results showed that cDNA microarray was a useful tool to study the molecular mechanisms of disease resistance in plants.
从籼稻品种中156与谷梅2构建的重组自交系（RIL）中筛选出一对近等基因系G205和G71。在抗稻瘟病基因座Pi-25(t)上，G205含有来自谷梅2的抗性等位基因，而G71含有来自中156的感病等位基因。就遗传背景而言，在检测的672个RFLP或SSLP位点中，仅在24个位点上检测到不同的等位基因。利用包含2200个表达序列标签（EST）的cDNA微阵列研究了G205和G71对稻瘟病菌的表达谱。在四叶期用病原菌接种叶片12小时，共鉴定出998个基因。仅在G205中有3个基因被真菌显著上调。其中两个基因的功能已知，但第三个基因的功能未知。这两个基因分别编码酪蛋白激酶IIα亚基和反转录转座子TOS17插入元件。另外35个基因在近等基因系间具有相似的表达模式。其中，17个基因被接种上调，18个基因被接种下调。35个基因中有33个基因的功能已知。BLAST分析表明，所有35个功能已知的基因都与防御反应、信号转导、胁迫响应、光合作用和糖代谢相关。Northern杂交证实，随机选择的5个差异表达基因中有4个与cDNA微阵列检测到的表达模式相同。其中两个上调基因分别编码酪蛋白激酶IIα亚基和富含甘氨酸蛋白（Grp），另外两个下调基因分别编码腈水解酶相关蛋白和18S小亚基核糖体RNA基因。Northern杂交还显示，接种真菌后0至36小时，Grp的表达持续上调。这些结果表明，cDNA微阵列是研究植物抗病分子机制的有用工具。
