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Identification and characterization of genes involved in the interaction between rice and rice blast fungus, Magnaporthe grisea.

机译：水稻和稻瘟病菌（Magnaporthe grisea）相互作用中涉及的基因的鉴定和表征。

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摘要

To better understand the molecular basis of the defense response against the rice blast fungus (Magnaporthe grisea), a large-scale expressed sequence tag (EST) sequencing approach was used to identify genes involved in the early infection stages in rice (Oryza sativa). Six cDNA libraries were constructed using infected leaf tissues harvested from 6 conditions: resistant, partially resistant, and susceptible reactions at both 6 and 24 h after inoculation. Two additional libraries were constructed using uninoculated leaves and leaves from the lesion mimic mutant spl11. A total of 68,920 ESTs were generated from 8 libraries. Clustering and assembly analyses resulted in 13,570 unique sequences from 10,934 contigs and 2,636 singletons. Gene function classification showed that 42% of the ESTs were predicted to have putative gene function. Comparison of the pathogen-challenged libraries with the uninoculated control library revealed an increase in the percentage of genes in the functional categories of defense and signal transduction mechanisms and cell cycle control, cell division, and chromosome partitioning. In addition, hierarchical clustering analysis grouped the eight libraries based on their disease reactions. A total of 7,748 new and unique ESTs were identified from our collection compared with the KOME full-length cDNA collection. Interestingly, we found that rice ESTs are more closely related to sorghum (Sorghum bicolor) ESTs than to barley (Hordeum vulgare ), wheat (Triticum aestivum), and maize ( Zea mays) ESTs. To date, this is the largest EST collection generated from a single plant-pathogen interaction in plants. Therefore, the sequences reported in this study provide a significant improvement in our understanding of the rice defense mechanism to the rice blast fungus and will streamline the community effort in elucidating the functions of many defense response genes in rice.; Ubiquitination-regulated protein degradation is a common regulatory mechanism that controls a range of cellular processes in eukaryotes. Recent studies showed that ubiquitination might also play an important role in plant disease resistance. We have recently identified 43 rice ubiquitination-related E2 and E3 genes in our microarray analysis. These genes were induced or suppressed in Pi2 and Pi9 resistant plants after rice blast inoculation. The expression patterns of these E2 and E3 genes were confirmed using northern blot and RT-PCR methods. Among these E2 and E3 genes, we chose the homologue (OsSINAT5) of the Arabidopsis SINAT5 for a detailed functional analysis. Northern blot hybridization and RT-PCR confirmed that OsSINAT5 was induced by rice blast infection as early as 6 hr after inoculation. Functional analysis of this gene in resistance to rice pathogens was conducted using reverse genetics and biochemical approaches.
机译：为了更好地理解针对稻瘟病菌（Magnaporthe grisea）的防御反应的分子基础，使用了大规模表达序列标签（EST）测序方法来鉴定参与水稻（Oryza sativa）早期感染阶段的基因。使用从6种条件下收获的受感染叶片组织构建了六个cDNA文库：在接种后6和24小时都产生了抗性，部分抗性和易感反应。使用未接种的叶片和来自病变模拟突变体spl11的叶片构建了另外两个文库。从8个文库中总共产生了68,920个EST。聚类和装配分析产生了来自10,934个重叠群和2,636个单例的13,570个独特序列。基因功能分类显示，预计42％的EST具有推定的基因功能。将病原体攻击的文库与未接种的对照文库进行比较，发现防御和信号转导机制以及细胞周期控制，细胞分裂和染色体分区等功能类别中的基因百分比有所增加。此外，分层聚类分析基于八个库的疾病反应将其分组。与KOME全长cDNA集合相比，从我们的集合中总共鉴定了7,748个新的和独特的EST。有趣的是，我们发现与大麦（Hordeum vulgare），小麦（Triticum aestivum）和玉米（Zea mays）的EST相比，水稻的EST与高粱（sorghum bicolor）的EST更为紧密。迄今为止，这是从植物中单一植物-病原体相互作用产生的最大的EST集合。因此，本研究报道的序列大大提高了我们对稻瘟病菌的水稻防御机制的了解，并将简化阐明水稻中许多防御反应基因功能的社区努力。泛素化调节的蛋白质降解是控制真核生物中一系列细胞过程的常见调节机制。最近的研究表明，泛素化在植物抗病性中也可能起重要作用。我们最近在我们的微阵列分析中鉴定了43个与水稻泛素化相关的E2和E3基因。稻瘟病接种后，在Pi2和Pi9抗性植物中诱导或抑制了这些基因。使用Northern印迹和RT-PCR方法证实了这些E2和E3基因的表达模式。在这些E2和E3基因中，我们选择了拟南芥SINAT5的同源物（OsSINAT5）进行详细的功能分析。 Northern印迹杂交和RT-PCR证实OsSINAT5早在接种后6小时就被稻瘟病感染诱导。使用反向遗传学和生化方法对该基因在水稻病原体抗性中的功能进行了分析。

著录项

  • 作者

    Jantasuriyarat, Chatchawan.;

  • 作者单位

    The Ohio State University.;

  • 授予单位 The Ohio State University.;
  • 学科 Biology Genetics.
  • 学位 Ph.D.
  • 年度 2006
  • 页码 115 p.
  • 总页数 115
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 遗传学;
  • 关键词

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