Study of the defense response and gene regulation for pathogen attack

本研究的目的是阐明植物主动防御反应基因的信号转导和调控机制。我们深入研究了豌豆PAL和CHS基因激活和失活的分子基础，特别是豌豆病原菌松顶球腔菌（Mycosphaerella pinodes）产生的激发子和抑制子的作用。1.通过对PAL基因的一个成员（PSPAL 1）的人工重建，以及在豌豆的一个多基因小家族中的功能获得和功能丧失实验，证实了Box 2和Box 4的保守序列基序在真菌诱导子诱导的PAL基因激活过程中起着重要作用。然而，Box 1与Box 2和Box 4以协同的方式起作用，可能形成转录激活复合物。2.携带嵌合PSPAL 1启动子-GUS-Nos终止子构建体的转基因烟草的产生（除了这些构建体连接到GUS报告基因而不是CAT基因之外，与1中所述的嵌合基因的类型相同）证明了框1、2和4，是响应病原体攻击的先决条件顺式元件。这些元件在响应由真菌诱导子处理介导的活化中也是重要的。-1，394至+140或至+115 PSPAL 1（翻译融合型）启动子贡献了组织特异性表达，特别是在根的分生组织、茎的下部中，以及在花器官的组织中非常高的表达，在花器官中粉红色色素沉积并且木质素被强烈地用于介导。394至+5或至-27的转基因烟草表现出非常少的GUS表达，表明外显子序列在转基因烟草的表达中是重要的。

The object of this research is to elucidate the mechanisms of signal transduction and control of the genes involved in plant active defense responses conferring host-pathogen specificity. We intensively characterized the molecular basis involved in the activation and deactivation of plant active defense genes encoding PAL and CHS in pea, particularly focused on the study of the effects of elicitor and suppressor produced by a pea pathogen, Mycosphaerella pinodes. Details of the current status of the studies are,1.Artificial reconstruction of a members of PAL gene (PSPAL1) and in a small multigene family of pea demonstrated that the conserved sequence motifs of Box 2 and Box 4 were very important in the activation mediated by the treatment with fungal elicitor by so called Loss of Function and Gain of Function Experiments. However, Box 1 functions in a concerted manner with Box 2 and Box 4, probably forming a transcriptional activation complex.2.Generation of transgenic tobacco carrying the chimeric PSPAL1 promoter-GUS-Nos terminator construct (same types of the chimeric gene as described in 1 except these construct are connected to GUS reporter gene instead of CAT gene) demonstrated that Boxes1,2, and 4, are prerequisite cis-elements in response to the pathogen attack. These elements are also important in response to the activation mediated by the treatment with fungal elicitor. -1,394 to +140 or to +115 of PSPAL1 (translational fusion type) promoter contributed the tissue-specific expression, particularly in the merismatic tissues of roots, lower part of stems and very high expression in the tissues of flower organs where pink pigment is deposited and lignins are vigorously for med. Chimeric fusion of -1,394 to +5 or to -27 exhibited very little GUS expression, indicating that the exonic sequences are important in the expression if transgenic tobacco.