[Identification of genes related to resistance to Magnaporthe grisea using differential display technique in rice].

Hai-Ying Zhang, Yong Liu, Dong-Cheng Liu, Xiu-Zhi Wang, Chao Wang, Ling-Xia Wang, Ai-Min Zhang, Ping Li
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Abstract

Rice blast caused by Magnaporthe grisea is one of the most serious constraints on high productivity. Understanding the mechanism of the infection of Magnaporthe grisea and the change of gene expression after infection is useful to control blast disease in rice. This work presents the isolation of differentially expressed cDNA fragments from rice leaf induced by the inoculum suspension of Magnaporthe grisea using mRNA differential display technique. Total 87 differential expressed cDNA fragments were recoveried and reamplified. The dot-blotting results showed that 6 fragments of 81 were confirmed to be the expression induced by Magnaporthe grisea inoculum. Those fragments were then cloned into vectors for sequencing. Sequence analysis through Internet Blast searching showed that 3 fragments were novel gene fragments. One was homologous with a putative malate synthase gene on rice chromosome 4 with 78% identities of amino acid; one was highly homologous (75% identity) with rice RPR1 gene on chromosome 11, which has a conservative structure of NBS-LRR domain and may be related to signal transduction of rice defense reaction;another one was homologous with a putative thioredoxin gene on rice chromosome 6 with the identity of 72%.

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[利用差异展示技术鉴定水稻抗稻瘟病相关基因]。
稻瘟病是制约水稻高产的最严重因素之一。了解稻瘟病的侵染机制及侵染后基因表达的变化,有助于水稻稻瘟病的防治。利用mRNA差异显示技术从稻瘟病病菌接种悬浮液诱导的水稻叶片中分离出差异表达的cDNA片段。共有87个差异表达的cDNA片段被恢复并重新扩增。点印迹结果表明,81个片段中有6个片段被证实是由稻瘟病病菌接种诱导的表达。然后将这些片段克隆到载体中进行测序。Internet Blast检索序列分析显示,其中3段为新基因片段。其中一个与水稻4号染色体上推测的苹果酸合成酶基因同源,氨基酸同源性为78%;一个与水稻11号染色体上的RPR1基因高度同源(同源度为75%),该基因具有NBS-LRR结构域的保守结构,可能与水稻防御反应的信号转导有关;另一个与水稻6号染色体上推定的硫氧还蛋白基因同源,同源度为72%。
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