Manthey Katja, Krajinski Franziska, Hohnjec Natalija, Firnhaber Christian, Pühler Alfred, Perlick Andreas M, Küster Helge
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Postfach 100131, D-33501 Bielefeld, Germany.
Mol Plant Microbe Interact. 2004 Oct;17(10):1063-77. doi: 10.1094/MPMI.2004.17.10.1063.
Transcriptome profiling based on cDNA array hybridizations and in silico screening was used to identify Medicago truncatula genes induced in both root nodules and arbuscular mycorrhiza (AM). By array hybridizations, we detected several hundred genes that were upregulated in the root nodule and the AM symbiosis, respectively, with a total of 75 genes being induced during both interactions. The second approach based on in silico data mining yielded several hundred additional candidate genes with a predicted symbiosis-enhanced expression. A subset of the genes identified by either expression profiling tool was subjected to quantitative real-time reverse-transcription polymerase chain reaction for a verification of their symbiosis-induced expression. That way, induction in root nodules and AM was confirmed for 26 genes, most of them being reported as symbiosis-induced for the first time. In addition to delivering a number of novel symbiosis-induced genes, our approach identified several genes that were induced in only one of the two root endosymbioses. The spatial expression patterns of two symbiosis-induced genes encoding an annexin and a beta-tubulin were characterized in transgenic roots using promoter-reporter gene fusions.
基于cDNA阵列杂交和计算机筛选的转录组分析被用于鉴定在苜蓿根瘤和丛枝菌根(AM)中均被诱导的蒺藜苜蓿基因。通过阵列杂交,我们分别检测到数百个在根瘤和AM共生中上调的基因,共有75个基因在两种相互作用中均被诱导。基于计算机数据挖掘的第二种方法产生了数百个额外的具有预测共生增强表达的候选基因。通过两种表达分析工具鉴定出的一部分基因进行了定量实时逆转录聚合酶链反应,以验证它们的共生诱导表达。通过这种方式,证实了26个基因在根瘤和AM中被诱导,其中大多数基因是首次被报道为共生诱导的。除了提供许多新的共生诱导基因外,我们的方法还鉴定出了几个仅在两种根内共生之一中被诱导的基因。使用启动子-报告基因融合技术在转基因根中对编码膜联蛋白和β-微管蛋白的两个共生诱导基因的空间表达模式进行了表征。