Identification of potato genes induced during colonization by Phytophthora infestans
Identification of potato genes induced during colonization by Phytophthora infestans
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DOI：
10.1046/j.1364-3703.2001.00059.x
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发表时间：
2001-05-01
影响因子：
4.9
通讯作者：
Collinge, Margaret
中科院分区：
文献类型：
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作者：
Beyer, Katinka;Binder, Andres;Collinge, Margaret
采用抑制减法杂交（Suppression Subtractive Hybridization， SSH）技术寻找马铃薯与疫霉亲和互作诱导的基因。以苯并（1,2,3）噻二唑-7-碳硫酸s-甲基lester （BTH）处理过的马铃薯叶片为对照组织，构建了一个低冗余、频率较低的经典植物致病相关（PR）基因文库。利用反北分析（将排列的克隆与放射性标记的cDNA群体杂交）对288个克隆进行诱导序列筛选。在用这种方法检测到的75个克隆中，有43个似乎是诱导的。然后对其中11个克隆进行了总RNA印迹分析，证实其中10个克隆在感染P. infestans期间转录物水平升高。RNA印迹分析分析的一些cdna与已知在感染期间诱导的基因具有同源性，例如与β -1,3-葡聚糖酶。另一组cdna与参与解毒的酶具有同源性：γ -谷氨酰半胱氨酸合成酶、细胞色素P450、谷胱甘肽s -转移酶和mrp型ABC转运蛋白。其他感染诱导的cdna编码先前未报道的感染诱导的推定蛋白质：例如，位于er的伴侣蛋白BiP，以及作为染色体分离突变抑制因子分离出来的细粒曲霉（Aspergillus nidulans SudD）的同源物。因此，差异文库提供了分析感染期间发生的代谢变化和更详细的疾病过程本身的机会。
Suppression Subtractive Hybridization (SSH) was applied in a search for genes induced during the compatible interaction between Phytophthora infestans and potato. Using potato leaves that had been treated with benzo(1,2,3) thiadiazole-7-carbothioic acid S-methylester (BTH) as the control tissue, a low redundancy library with a relatively low frequency of the classic plant Pathogenesis-Related (PR) genes was generated. 288 of the clones were screened for induced sequences using Inverse Northern analysis (hybridizing the arrayed clones with radiolabelled cDNA populations). Of the 75 clones that were detectable by this method, 43 appeared to be induced. Eleven of these clones were then analysed by total RNA blot analysis, and elevation of transcript levels during P. infestans infection was confirmed for 10 of them. Some of the cDNAs analysed by RNA blot analysis have homology to genes already known to be induced during infection, e. g. to beta-1,3-glucanase. Another group of cDNAs have homology to enzymes involved in detoxification: gamma-glutamylcysteine synthetase, cytochrome P450, glutathione S-transferase and an MRP-type ABC transporter. Other infection induced cDNAs encode putative proteins that have not previously been reported to be induced by infection: e. g. the ER-located chaperone BiP, and a homologue of Aspergillus nidulans SudD, which was isolated as a suppressor of a mutation in chromosome disjunction. The differential library therefore presents the opportunity to analyse the metabolic changes occurring during infection, and the disease process itself in more detail.