Abstract
BackgroundThe 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.ResultsTwelve 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 × 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.ConclusionThis 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.
Highlights
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
(page number not for citation purposes) http://www.biomedcentral.com/1471-2229/7/8 for three different tissues and 10 different stages of Pierce's disease development were constructed to identify spatial and temporal transcriptional changes resulting from X. fastidiosa infection
Out of the 1,942 non-redundant expressed sequence tags (EST) that were cloned in this study, about 33% were found to be unique, demonstrating the effectiveness of the experimental design and the construction strategy utilized for these suppression hybridization (SSH) libraries
Summary
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. Pierce's disease (PD) has been a chronic problem for California's grape industry since the 1880s. The threat from this disease has recently become more severe with the introduction and establishment of a more effective vector, the glassy-winged sharpshooter (Homalodisca coagulate). The disease is caused by Xylella fastidiosa, a xylem-limited, gram negative bacterium that is hosted by a wide range of plant species in and around vineyards in the southern United States and Mexico [1]. Even low populations of the glassy-winged sharpshooter can have severe impact on vineyard health, limiting the effectiveness of predators to solve PD. As pesticide use becomes more restricted and as pesticide resistance develops, it is likely that the ultimate solution to PD will be host resistance
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