Bastiaanse Héloïse, Muhovski Yordan, Parisi Olivier, Paris Roberta, Mingeot Dominique, Lateur Marc
Life Sciences Department, Breeding and Biodiversity Unit, Walloon Agricultural Research Center, Rue de Liroux, 4, 5030 Gembloux, Belgium.
BMC Genomics. 2014 Nov 29;15:1043. doi: 10.1186/1471-2164-15-1043.
Scab, caused by the fungus Venturia inaequalis, is one of the most important diseases of cultivated apple. While a few scab resistance genes (R genes) governing qualitative resistance have been isolated and characterized, the biological roles of genes governing quantitative resistance, supposed to be more durable, are still unknown. This study aims to investigate the molecular mechanisms involved in the partial resistance of the old Belgian apple cultivar 'Président Roulin' against V. inaequalis.
A global gene expression analysis was conducted in 'Président Roulin' (partially resistant) and in 'Gala' (susceptible) challenged by V. inaequalis by using the cDNA-AFLP method (cDNA-Amplified Fragment Length Polymorphism). Transcriptome analysis revealed significant modulation (up- or down-regulation) of 281 out of approximately 20,500 transcript derived fragments (TDFs) in 'Président Roulin' 48 hours after inoculation. Sequence annotation revealed similarities to several genes encoding for proteins belonging to the NBS-LRR and LRR-RLK classes of plant R genes and to other defense-related proteins. Differentially expressed genes were sorted into functional categories according to their gene ontology annotation and this expression signature was compared to published apple cDNA libraries by Gene Enrichment Analysis. The first comparison was made with two cDNA libraries from Malus x domestica uninfected leaves, and revealed in both libraries a signature of enhanced expression in 'Président Roulin' of genes involved in response to stress and photosynthesis. In the second comparison, the pathogen-responsive TDFs from the partially resistant cultivar were compared to the cDNA library from inoculated leaves of Rvi6 (HcrVf2)-transformed 'Gala' lines (complete disease resistance) and revealed both common physiological events, and notably differences in the regulation of defense response, the regulation of hydrolase activity, and response to DNA damage. TDFs were in silico mapped on the 'Golden Delicious' apple reference genome and significant co-localizations with major scab R genes, but not with quantitative trait loci (QTLs) for scab resistance nor resistance gene analogues (RGAs) were found.
This study highlights possible candidate genes that may play a role in the partial scab resistance mechanisms of 'Président Roulin' and increase our understanding of the molecular mechanisms involved in the partial resistance against apple scab.
由不等黑星菌(Venturia inaequalis)引起的苹果黑星病是栽培苹果最重要的病害之一。虽然已经分离和鉴定了一些控制质量抗性的黑星病抗性基因(R基因),但控制数量抗性(据推测更持久)的基因的生物学作用仍然未知。本研究旨在探究古老的比利时苹果品种‘鲁林总统’对不等黑星菌的部分抗性所涉及的分子机制。
采用cDNA-AFLP方法(cDNA扩增片段长度多态性),对受不等黑星菌侵染的‘鲁林总统’(部分抗性)和‘嘎啦’(感病)进行了全基因组基因表达分析。转录组分析显示,接种48小时后,‘鲁林总统’中约20,500个转录本衍生片段(TDF)中有281个发生了显著调控(上调或下调)。序列注释显示,这些片段与几个编码属于植物R基因的NBS-LRR和LRR-RLK类蛋白质的基因以及其他防御相关蛋白质相似。根据基因本体注释,将差异表达基因分类到功能类别中,并通过基因富集分析将这种表达特征与已发表的苹果cDNA文库进行比较。第一次比较是与来自未感染叶片的两个苹果(Malus x domestica)cDNA文库进行的,结果显示在两个文库中,‘鲁林总统’中参与应激反应和光合作用的基因均有增强表达的特征。在第二次比较中,将部分抗性品种中对病原体有反应的TDF与Rvi6(HcrVf2)转化的‘嘎啦’株系(完全抗病)接种叶片的cDNA文库进行比较,结果显示既有共同的生理事件,特别是在防御反应调控、水解酶活性调控和对DNA损伤的反应方面存在差异。通过电子定位将TDF定位到‘金冠’苹果参考基因组上,未发现与主要黑星病R基因有显著共定位,但与黑星病抗性数量性状位点(QTL)或抗性基因类似物(RGA)无共定位。
本研究突出了可能在‘鲁林总统’的部分黑星病抗性机制中发挥作用的候选基因,并增进了我们对苹果黑星病部分抗性所涉及分子机制的理解。