Bacterial surface appendages called pili and needle-like filaments are associated with protein and/or DNA transfer to recipient plant, human, or bacterial cells during pathogenesis or conjugation. Although it has long been suspected that pili function as a conduit for protein or DNA transfer, direct evidence has been lacking. The Hrp pilus of Pseudomonas syringae is assembled by the type III secretion system. We used an in situ immunogold labeling procedure to visualize the extrusion of an effector protein, AvrPto, from the tip of the Hrp pilus, providing direct evidence that a bacterial pilus can function as a conduit for protein delivery.
Bacterial surface appendages called pili often are associated with DNA and͞or protein transfer between cells. The exact function of pili in the transfer process is not understood and is a matter of considerable debate. The Hrp pilus is assembled by the Hrp type III protein secretion system of Pseudomonas syringae pv. tomato (Pst) strain DC3000. In this study, we show that the hrpA gene, which encodes the major subunit of the Hrp pilus, is required for secretion of putative virulence proteins, such as HrpW and AvrPto. In addition, the hrpA gene is required for full expression of genes that encode regulatory, secretion, and effector proteins of the type III secretion system. hrpA-mediated gene regulation apparently is through effect on the mRNA level of two previously characterized regulatory genes, hrpR and hrpS. Ectopic expression of the hrpRS gene operon restored gene expression, but not protein secretion, in the hrpA mutant. Three single amino acid mutations at the HrpA carboxyl terminus were identified that affect the secretion or regulatory function of the HrpA protein. These results define an essential role of the Hrp pilus structural gene in protein secretion and coordinate regulation of the type III secretion system in Pst DC3000.
The NPR1 gene plays a pivotal role in systemic acquired resistance in plants. Its overexpression in Arabidopsis and rice results in increased disease resistance and elevated expression of pathogenesis-related (PR) genes. An NPR1 homolog, MpNPR1-1, was cloned from apple (Malus x domestica) and overexpressed in two important apple cultivars, Galaxy and M26. Apple leaf pieces were transformed with the MpNPR1 cDNA under the control of the inducible Pin2 or constitutive Cauliflower mosaic virus (CaMV)35S promoter using Agrobacterium tumefaciens. Overexpression of MpNPR1 mRNA was shown by reverse transcriptase-polymerase chain reaction. Activation of some PR genes (PR2, PR5, and PR8) was observed. Resistance to fire blight was evaluated in a growth chamber by inoculation of the shoot tips of our own rooted 30-cm-tall plants with virulent strain Ea273 of Erwinia amylovora. Transformed Galaxy lines overexpressing MpNPR1 had 32 to 40% of shoot length infected, compared with 80% in control Galaxy plants. Transformed M26 lines overexpressing MpNPR1 under the control of the CaMV35S promoter also showed a significant reduction of disease compared with control M26 plants. Some MpNPR-overexpressing Galaxy lines also exhibited increased resistance to two important fungal pathogens of apple, Venturia inaequalis and Gymnosporangium juniperi-virginianae. Selected transformed lines have been propagated for field trials for disease resistance and fruit quality.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.