Lin Hong, Doddapaneni Harshavardhan, Takahashi Yuri, Walker M Andrew
USDA-ARS, 9611 S, Riverbend Avenue, Parlier, California 93648, USA.
BMC Plant Biol. 2007 Feb 22;7:8. doi: 10.1186/1471-2229-7-8.
The gram-negative bacterium Xylella fastidiosa (Xf) is the causal agent of Pierce's disease (PD) in grape as well as diseases of many fruit and ornamental plants. The current molecular breeding efforts have identified genetic basis of PD resistance in grapes. However, the transcriptome level characterization of the host response to this pathogen is lacking.
Twelve tissue specific subtractive suppression hybridization (SSH) cDNA libraries derived from a time course sampling scheme were constructed from stems, leaves and shoots of PD resistant and susceptible sibling genotypes (V. rupestris x V. arizonica) in response to Xf infection. A total of 5,794 sequences were obtained from these cDNA libraries from which 993 contigs and 949 singletons were derived. Using Gene Ontology (GO) hierarchy, the non-redundant sequences were classified into the three principal categories: molecular function (30%), cellular components (9%) and biological processes (7%). Comparative analysis found variations in EST expression pattern between infected and non-infected PD resistant and PD susceptible grape genotypes. Among the three tissues, libraries from stem tissues showed significant differences in transcript quality suggesting their important role in grape-Xylella interaction.
This study constitutes the first attempt to characterize the Vitis differential transcriptome associated with host-pathogen interactions from different explants and genotypes. All the generated ESTs have been submitted to GenBank and are also available through our website for further functional studies.
革兰氏阴性菌木质部难养菌(Xylella fastidiosa,Xf)是葡萄上皮尔氏病(Pierce's disease,PD)以及许多果树和观赏植物疾病的病原体。目前的分子育种工作已确定了葡萄中抗PD的遗传基础。然而,宿主对这种病原体反应的转录组水平特征尚不清楚。
从抗PD和感病的同胞基因型(河岸葡萄x亚利桑那葡萄)的茎、叶和嫩梢中,根据时间进程采样方案构建了12个组织特异性消减抑制杂交(SSH)cDNA文库,以响应Xf感染。从这些cDNA文库中总共获得了5794个序列,从中得到了993个重叠群和949个单拷贝序列。利用基因本体论(GO)层次结构,将非冗余序列分为三大类:分子功能(30%)、细胞成分(9%)和生物学过程(7%)。比较分析发现,受感染和未受感染的抗PD和感病葡萄基因型之间的EST表达模式存在差异。在这三种组织中,茎组织文库的转录本质量存在显著差异,表明它们在葡萄与木质部难养菌相互作用中起重要作用。
本研究首次尝试从不同外植体和基因型中表征与宿主-病原体相互作用相关的葡萄差异转录组。所有生成的EST已提交至GenBank,也可通过我们的网站获取,以供进一步的功能研究使用。