林木转基因抗病存在广谱抗性、持久抗性和低转化率等困难。设计人工病原物诱导启动子(SPIP)调控SWAP70表达，激活信号网络的关键节点Rop，促进尾叶桉芽分化并使其具广谱抗病性。以GWSF-mini35S为基础，优化元件类型、排列、间隔序列等，获得理想的SPIP，建立序列设计与SPIP转录特性关系的数学模型。克隆SWAP70基因，构建SPIP调控SWAP70表达的载体，转化尾叶桉。克隆Rop基因，通过酵母双杂交筛选与SWAP70互作、参与抗病反应或影响愈伤分化的Rop蛋白。分别在愈伤分化和植株生长阶段，通过信号分子类型和浓度的变化诱导SPIP，调控SWAP70的表达，影响Rop活性，检测激素含量、ROS含量、相关酶活性及基因的表达变化，探讨Rop与愈伤分化和抗病间的信号互作。本研究是林木转基因广谱抗病的新尝试，可为SPIP的设计提供参考，为阐明桉树Rop抗病和愈伤分化信号网络积累资料。

To promote callus differentiation and confer broad-spectrum disease resistance on Eucalyptus Urophylla, synthetic pathogen-inducible promotors (SPIPs) are designed to regulate SWAP70 expression. Rho-related GTPase of plants (Rop), a key GTPases involved in diverse signaling processes including immunity, growth and development, could be actived by interacting with SWAP70, which replaces bound GDP by GTP converting the molecular switch to the active conformation. .GWSF-mini35S promotor with low background transcriptional activity, screened from 8 SPIPs at previous study, could be induced by salicylic acid, Ralstonia solanacearum and the spores of Phytophthora capsici. At the basis of GWSF-mini35S promotor, the factors influencing the transcriptional character of SPIPs will be optimized, including cis-element types, arrangement, location, etc. And the ideal SPIP, which achieves some advantages, such as more induction factors, low basal activity, high expression activity and quick expression initiation, will be selected to subsequent study. Using the results of SPIP transcriptional character as foundation and consulting the transcriptional character of pathogen-inducible promoters reported at published literature, the mathematical model concerning the relationship between transcriptional character and the sequence of pathogen-inducible promoters would be established by genetic algorithm or simulated annealing algorithm. .The cDNA sequences of Arabidopsis SWAP70 gene and Eucalyptus SWAP70 are amplified from leaf cDNA, respectively. Then, the SWAP70 genes are used to construct plant inducible expression vector under the control of optimized SPIP. The genes are introduced into E. Urophylla by Agrobacterium tumefaciens mediated transformation. And the stable transformations are confirmed by PCR and Southern blot. .The interactions between SWAP70 and Ropx in callus differentiation and disease resistance of E. Urophylla are studied by yeast two-hybrid system. The cloned SWAP70 genes are used to construct bait voctor. The full-length Rop genes are isolated from Eucalyptus by PCR-RACE and they are used to construct prey voctors..In order to evaluate the effect on differentiation of adventitious buds of SWAP70 differential expression, it was induced by different inductor at callus differentiation stage of E. Urophylla. The content of hormone and reactive oxygen species (ROS) and expression changes of related enzymes are detected..Regulated SWAP70 expression with several pathogenic factors at growth stage of transgenic plant, the content of ROS, the expressional level of pathogenesis-related proteins and related enzymatic activities are detected. .The project is a new trials of tree broad-spectrum resistance by genetic engineering. It will offer helpful references to the design of SPIP and accumulate experiment evidence to clarify Rop signal network in Eucalyptus.