Isolation and characterization of broad-spectrum disease-resistant Arabidopsis mutants.
Isolation and characterization of broad-spectrum disease-resistant Arabidopsis mutants.
复制标题
广谱抗病拟南芥突变体的分离和表征。
DOI：
10.1093/genetics/160.4.1661
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发表时间：
2002
期刊：
影响因子：
3.3
通讯作者：
Ryals,JohnA
中科院分区：
文献类型：
--
作者：
Maleck,Klaus;Neuenschwander,Urs;Cade,RebeccaM;Dietrich,RobertA;Dangl,JefferyL;Ryals,JohnA
为了鉴定组成型表达系统获得抗性（SAR）的拟南芥突变体，我们构建了在SAR诱导的PR-1启动子（PR-1/luc）控制下表达萤火虫荧光素酶基因的报告基因系。在EMS诱变一个良好表征的转基因株系后，我们筛选了250，000株M2植物体内报告基因的组成型表达。从一个突变体的集合含有几百个假定的突变体，我们集中在16个突变体缺乏自发性过敏反应（HR）细胞死亡。我们将其中4个组成性免疫（cim）突变体定位到染色体臂上。通过分析对寄生霜霉和假单胞菌菌株的反应，通过对内源标记基因的RNA印迹分析，以及通过测定突变体中的水杨酸水平，建立了抗病性的组成型表达。这些表型的多样性使我们能够在典型的SAR信号传导途径和一个单独的途径中定义不同的步骤，该途径仅在突变体的一个子集中具有活性。
To identify Arabidopsis mutants that constitutively express systemic acquired resistance (SAR), we constructed reporter lines expressing the firefly luciferase gene under the control of the SAR-induciblePR-1promoter (PR-1/luc). After EMS mutagenesis of a well-characterized transgenic line, we screened 250,000 M2plants for constitutive expression of the reporter genein vivo. From a mutant collection containing several hundred putative mutants, we concentrated on 16 mutants lacking spontaneous hypersensitive response (HR) cell death. We mapped 4 of theseconstitutiveimmunity (cim) mutants to chromosome arms. Constitutive expression of disease resistance was established by analyzing responses to virulentPeronospora parasiticaandPseudomonas syringaestrains, by RNA blot analysis for endogenous marker genes, and by determination of salicylic acid levels in the mutants. The variety of thecimphenotypes allowed us to define distinct steps in both the canonical SAR signaling pathway and a separate pathway for resistance toErysiphe cichoracearum, active in only a subset of the mutants.