Ramonell Katrina, Berrocal-Lobo Marta, Koh Serry, Wan Jinrong, Edwards Herb, Stacey Gary, Somerville Shauna
Department of Biological Sciences, University of Alabama, Tuscaloosa, Alabama 35487-0344, USA.
Plant Physiol. 2005 Jun;138(2):1027-36. doi: 10.1104/pp.105.060947. Epub 2005 May 27.
Chitin is a major component of fungal walls and insect exoskeletons. Plants produce chitinases upon pathogen attack and chito-oligomers induce defense responses in plants, though the exact mechanism behind this response is unknown. Using the ATH1 Affymetrix microarrays consisting of about 23,000 genes, we examined the response of Arabidopsis (Arabidopsis thaliana) seedlings to chito-octamers and hydrolyzed chitin after 30 min of treatment. The expression patterns elicited by the chito-octamer and hydrolyzed chitin were similar. Microarray expression profiles for several genes were verified via northern analysis or quantitative reverse transcription-PCR. We characterized T-DNA insertion mutants for nine chito-oligomer responsive genes. Three of the mutants were more susceptible to the fungal pathogen, powdery mildew, than wild type as measured by conidiophore production. These three mutants included mutants of genes for two disease resistance-like proteins and a putative E3 ligase. The isolation of loss-of-function mutants with enhanced disease susceptibility provides direct evidence that the chito-octamer is an important oligosaccharide elicitor of plant defenses. Also, this study demonstrates the value of microarray data for identifying new components of uncharacterized signaling pathways.
几丁质是真菌细胞壁和昆虫外骨骼的主要成分。植物在受到病原体攻击时会产生几丁质酶,几丁质寡聚物会诱导植物产生防御反应,不过这种反应背后的确切机制尚不清楚。我们使用由约23000个基因组成的ATH1 Affymetrix微阵列,检测了拟南芥(Arabidopsis thaliana)幼苗在处理30分钟后对几丁质八聚体和水解几丁质的反应。几丁质八聚体和水解几丁质引发的表达模式相似。通过Northern分析或定量逆转录PCR验证了几个基因的微阵列表达谱。我们对9个几丁质寡聚物响应基因的T-DNA插入突变体进行了表征。通过分生孢子产生量测定,其中3个突变体比野生型对真菌病原体白粉病更敏感。这3个突变体包括两个抗病样蛋白基因和一个推定的E3连接酶基因的突变体。具有增强病害易感性的功能缺失突变体的分离提供了直接证据,表明几丁质八聚体是植物防御的重要寡糖激发子。此外,这项研究证明了微阵列数据在鉴定未表征信号通路新成分方面的价值。