title: Expression of chloroperoxidase from Pseudomonas pyrrocinia in tobacco plastids for fungal resistance.
abstract: The chloroperoxidase (cpo) gene from Pseudomonas pyrrocinia was transformed into the plastid genome (plastome) of Nicotiana tabacum var. Petit Havana and transplastomic lines were compared with a nuclear transformant for the same gene. Southern analysis confirmed integration in the plastome and western blotting confirmed the presence of the chloroperoxidase protein (CPO) in higher abundance in transplastomic plants than in cpo nuclear transformants. Northern analysis of primary plastome transformants for cpo showed 15-fold higher transcript abundance than in the nuclear transformant, yet this extent of enhancement was not observed in western blot, enzyme or bioassay, indicating a bottleneck at the post-transcriptional level. Representative plants from the two transplastomic lines showed resistance to fungal pathogens in vitro (Aspergillus flavus, Fusarium verticillioides, and Verticillium dahliae) and in planta (Alternaria alternata).
authors.affiliation: USDA, ARS, Southern Regional Research Center, 1100 Robert E. Lee Boulevard, New Orleans, LA 70124-4305, United States. Electronic address: truhlman@austin.utexas.edu.
authors.forename: Tracey A
authors.initials: TA
authors.lastname: Ruhlman
authors.affiliation: USDA, ARS, Southern Regional Research Center, 1100 Robert E. Lee Boulevard, New Orleans, LA 70124-4305, United States. Electronic address: Rajah.Rajasekaran@ars.usda.gov.
authors.forename: Kanniah
authors.initials: K
authors.lastname: Rajasekaran
authors.affiliation: USDA, ARS, Southern Regional Research Center, 1100 Robert E. Lee Boulevard, New Orleans, LA 70124-4305, United States. Electronic address: Jeff.Cary@ars.usda.gov.
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Cary
mesh_terms: D002711:Chloride Peroxidase / Q000235:genetics / Q000378:metabolism*; D060467:Disease Resistance / Q000235:genetics*; D005658:Fungi / Q000502:physiology*; D054884:Host-Pathogen Interactions; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology; D018087:Plastids / Q000201:enzymology; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology* / Q000382:microbiology
chemical_list: D010940:Plant Proteins; D012333:RNA, Messenger; D002711:Chloride Peroxidase
keywords: Antimicrobial proteins; Chloroplast transformation; Disease resistance; Peroxidase; Transgene expression; Untranslated region