Gioti A, Simon A, Le Pêcheur P, Giraud C, Pradier J M, Viaud M, Levis C
Unité P.M.D.V, I.N.R.A, Route de St-Cyr, 78026 Versailles, France.
J Mol Biol. 2006 Apr 28;358(2):372-86. doi: 10.1016/j.jmb.2006.01.076. Epub 2006 Feb 7.
The ascomycete Botrytis cinerea is a broad-spectrum plant pathogen. Here, we describe the first macroarray transcriptomic study of the fungus in real-time infection conditions. Infection of Arabidopsis thaliana leaves by B.cinerea was monitored using macroarrays, containing 3032 genes. Variance analysis revealed that 7% of B.cinerea genes are differentially expressed during infection and allowed us to identify 27 genes significantly up-regulated in planta. Among them, two genes have already been associated with fungal pathogenicity, while eight genes have unidentified functions. The 27 genes were separated into three groups according to their expression profile. The first group showed maximal expression at the early stage following fungal penetration, the second one showed maximal expression at the outset of the colonization of plant leaves and the third group showed maximal expression when the colonization of plant leaves was completed. A gene of the last group (BcPIC5), which is homologous to FKBP12 proteins, was disrupted in order to determine its role in pathogenicity. At seven days post-inoculation, the lesions caused by the DeltaBcPIC5 mutant on bean leaves were reduced by 69% and did not further expand compared to the wild-type. These results confirm that transcriptomic analysis under infection conditions can be very valuable for the identification of fungal genes related to pathogenicity.
子囊菌灰葡萄孢是一种广谱植物病原体。在此,我们描述了该真菌在实时感染条件下的首次宏阵列转录组学研究。使用包含3032个基因的宏阵列监测灰葡萄孢对拟南芥叶片的感染。方差分析表明,7%的灰葡萄孢基因在感染过程中差异表达,这使我们能够鉴定出27个在植物体内显著上调的基因。其中,两个基因已与真菌致病性相关,而八个基因功能未知。根据表达谱,这27个基因被分为三组。第一组在真菌穿透后的早期阶段表达量最高,第二组在植物叶片定殖开始时表达量最高,第三组在植物叶片定殖完成时表达量最高。为了确定最后一组中的一个与FKBP12蛋白同源的基因(BcPIC5)在致病性中的作用,对其进行了敲除。接种七天后,与野生型相比,ΔBcPIC5突变体在菜豆叶片上引起的病斑减少了69%,且不再进一步扩展。这些结果证实,感染条件下的转录组分析对于鉴定与致病性相关的真菌基因非常有价值。