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Title: INTRODUCTION OF FOREIGN DNA INTO XYLELLA FASTIDIOSA

Author
item QIN, XIAOTING - VISITING SCIENTIST
item Hartung, John

Submitted to: International Society of Citriculture Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 12/7/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Citrus variegated chlorosis disease can cause dramatic yield loss and currently affects 30% of the sweet orange trees grown in Sao Paulo State, Brazil. It is spread both by contaminated budwood and by a variety of leaf hopper insect vectors, closely related to insects present in the United States. Once introduced into a grove, there are no effective control measures for the disease. Genetic analysis of the pathogen may provide insights leading to effective control strategies. We have constructed a plasmid vector based on a plasmid that occurs naturally in strain LAR-20 of X. fastidiosa and introduced it into two strains of X. fastidiosa by electroporation. Transformants acquired resistance to carbenicillin and the presence of the foreign DNA was confirmed by hybridization analysis. The transformants were confirmed to be X. fastidiosa by PCR tests specific for that bacterium. We have also used a three parent conjugational mating system to introduce the green fluorescent protein marker into strain LAR-20. Recombinant strains expressed GFP, were resistant to kanamycin, and the presence of the gfp gene was confirmed by PCR analysis. This marker may be useful for in planta studies of bacterial movement. Transconjugants were confirmed to be X. fastidiosa by PCR. X. fastidiosa is now amenable to genetic analysis which will exploit the genomic sequence information obtained in Brazil.