Open AccessDiagnosis of Pierce's Disease Using Biomarkers Specific to Xylella fastidiosa rRNA and Vitis vinifera Gene Expression
Author(s) -
Hyunju Choi,
F. Goes da Silva,
Hyunju Lim,
Alberto Iandolino,
Yunhee Seo,
S.-W. Lee,
Douglas R. Cook
Publication year - 2010
Publication title -
phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.264
H-Index - 131
eISSN - 1943-7684
pISSN - 0031-949X
DOI - 10.1094/phyto-01-10-0014
Subject(s) - xylella fastidiosa , biology , gene , biomarker , ribosomal rna , vitis vinifera , phytoplasma , polymerase chain reaction , genetics , computational biology , botany , bacteria , restriction fragment length polymorphism
Pierce's disease (PD), caused by Xylella fastidiosa, represents one of the most damaging diseases of cultivated grape. Management of PD in the vineyard often relies on the removal of infected individuals, which otherwise serve as a source of inoculum for nearby healthy vines. Effective implementation of such control measures requires early diagnosis, which is complicated by the fact that infected vines often harbor high titers of the pathogen in advance of visual symptom development. Here, we report a biomarker system that simultaneously monitors Xylella-induced plant transcripts as well as Xylella ribosomal (r)RNA. Plant biomarker genes were derived from a combination of in silico analysis of grape expressed sequence tags and validation by means of reverse-transcriptase polymerase chain reaction (RT-PCR). Four genes upregulated upon PD infection were individually multiplexed with an X. fastidiosa marker rRNA and scored using either real-time RT-PCR or gel-based conventional RT-PCR techniques. The system was sufficiently sensitive to detect both host gene transcript and pathogen rRNA in asymptomatic infected plants. Moreover, these plant biomarker genes were not induced by water deficit, which is a component of PD development. Such biomarker genes could have utility for disease control by aiding early detection and as a screening tool in breeding programs.