Comparative analysis of ESTs involved in grape responses to Xylella fastidiosa infection.
BACKGROUND
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.


RESULTS
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.


CONCLUSIONS
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.
Lin
Hong
H
USDA-ARS, 9611 S, Riverbend Avenue, Parlier, California 93648, USA. hlin@fresno.ars.usda.gov
Doddapaneni
Harshavardhan
H
Takahashi
Yuri
Y
Walker
M Andrew
MA
D016000:Cluster Analysis; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology; D018520:Plant Shoots / Q000235:genetics / Q000382:microbiology; D018547:Plant Stems / Q000235:genetics / Q000382:microbiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013997:Time Factors; D027843:Vitis / Q000235:genetics* / Q000382:microbiology; D044167:Xylella / Q000254:growth & development*
D018076:DNA, Complementary