Golkari Saber, Gilbert Jeannie, Prashar Suvira, Procunier J Douglas
Cereal Research Centre, Agriculture & Agri-Food Canada, 195 Dafoe Road, Winnipeg, MB, R3T 2M9, Canada.
Plant Biotechnol J. 2007 Jan;5(1):38-49. doi: 10.1111/j.1467-7652.2006.00213.x.
A wheat cDNA microarray consisting of 5739 expressed sequence tags (ESTs) was used to investigate the transcriptome patterns of the glume, lemma, palea, anther, ovary and rachis dissected from infected wheat spikes after inoculation with the fungus Fusarium graminearum, the causal agent of fusarium head blight (FHB) disease. Stringent conditions were employed to reduce the false discovery rate. The significance analysis of microarrays (SAM) was used to identify transcripts that showed a differential response between fungal-challenged vs. control plants. To verify the microarray data, Northern blot analysis was carried out on randomly selected up-regulated clones. We observed 185 (3.2%) up-regulated and 16 (0.28%) down-regulated ESTs in the six organs constituting the wheat spike. Many up-regulated ESTs (46.67%) showed no homology with sequences of known functions, whereas others showed homology with genes involved in defence and stress responses, the oxidative burst of H(2)O(2), regulatory functions, protein synthesis and the phenylpropanoid pathway. The monitoring of genes in specific organs avoided the averaging of expression values over multiple organs that occurs when using data from the whole spike. Our data allowed us to uncover new up-regulated genes expressed in specific organs. The study revealed that each organ had a defined and distinctive transcriptome pattern in response to F. graminearum infection.
使用包含5739个表达序列标签（EST）的小麦cDNA微阵列，来研究接种了镰刀菌属禾谷镰刀菌（引起赤霉病（FHB）的病原体）后，从受感染小麦穗上解剖下来的颖片、外稃、内稃、花药、子房和穗轴的转录组模式。采用严格条件以降低错误发现率。利用微阵列显著性分析（SAM）来鉴定在真菌挑战植株与对照植株之间表现出差异反应的转录本。为了验证微阵列数据，对随机选择的上调克隆进行了Northern印迹分析。我们在构成小麦穗的六个器官中观察到185个（3.2%）上调的EST和16个（0.28%）下调的EST。许多上调的EST（46.67%）与已知功能序列没有同源性，而其他EST与参与防御和应激反应、H₂O₂的氧化爆发、调节功能、蛋白质合成和苯丙烷途径的基因具有同源性。对特定器官中的基因进行监测避免了使用来自整个穗的数据时多个器官表达值的平均化。我们的数据使我们能够发现特定器官中表达的新的上调基因。该研究表明，每个器官在响应禾谷镰刀菌感染时都有明确且独特的转录组模式。
