• Molecules and Cells
• /
• v.25 no.1
• /
• pp.30-42
• /
• 2008

The disease-specific (dsp) region and the hypersensitive response and pathogenicity (hrp) genes, including the hrpW, $hrpN_{Ep}$, and hrpC operons have previously been sequenced in Erwinia pyrifoliae WT3 [Shrestha et al. (2005a)]. In this study, the remaining hrp genes, including the hrpC, hrpA, hrpS, hrpXY, hrpL and hrpJ operons, were determined. The hrp genes cluster (ca. 38 kb) was comprised of eight transcriptional units and contained nine hrc (hrp conserved) genes. The genetic organization of the hrp/hrc genes and their orientation for the transcriptions were also similar to and collinear with those of E. amylovora, showing ${\geq}80%$ homologies. However, ORFU1 and ORFU2 of unknown functions, present between the hrpA and hrpS operons of E. amylovora, were absent in E. pyrifoliae. To determine the HR active domains, several proteins were prepared from truncated fragments of the N-terminal and the C-terminal regions of $HrpN_{Ep}$ protein of E. pyrifoliae. The proteins prepared from the N-terminal region elicited HR, but not from those of the C-terminal region indicating that HR active domains are located in only N-terminal region of the $HrpN_{Ep}$ protein. Two synthetic oligopeptides produced HR on tobacco confirming presence of two HR active domains in the $HrpN_{Ep}$. The HR positive N-terminal fragment ($HN{\Delta}C187$) was further narrowed down by deleting C-terminal amino acids and internal amino acids to investigate whether amino acid insertion region have role in faster and stronger HR activity in $HrpN_{Ep}$ than $HrpN_{Ea}$. The $HrpN_{Ep}$ mutant proteins $HN{\Delta}C187$ (D1AIR), $HN{\Delta}C187$ (D2AIR) and $HN{\Delta}C187$ (DM41) retained similar HR activation to that of wild-type $HrpN_{Ep}$. However, the $HrpN_{Ep}$ mutant protein $HN{\Delta}C187$ (D3AIR) lacking third amino acid insertion region (102 to 113 aa) reduced HR when compared to that of wild-type $HrpN_{Ep}$. Reduction in HR elicitation could not be observed when single amino acids at different positions were substituted at third amino acids insertion region. But, substitution of amino acids at L103R, L106K and L110R showed reduction in HR activity on tobacco suggesting their importance in activation of HR faster in the $HrpN_{Ep}$ although it requires further detailed analysis.

• Molecules and Cells
• /
• v.24 no.2
• /
• pp.232-239
• /
• 2007

$HrpN_{EP}$, from the gram-negative pathogen, Erwinia pyrifoliae, is a member of the harpin group of proteins, inducing pathogen resistance and hypersensitive cell death in plants. When the $hrpN_{EP}$ gene driven by the OsCc1 promoter was introduced into tobacco plants via Agrobacterium-mediated transformation, their resistance to the necrotrophic fungal pathogen, Botrytis cinerea, increased. Resistance to B. cinerea was correlated with enhanced induction of SA-dependent genes such as PR-1a, PR2, PR3 and Chia5, of JA-dependent genes such as PR-1b, and of genes related to ethylene production, such as NT-EFE26, NT-1A1C, DS321, NT-ACS1 and NT-ACS2. However the expression of NPR1, which is thought to be essential for multiple-resistance, did not increase. Since the pattern of expression of defense-related genes in $hrpN_{EP}$-expressing tobacco differed from that in plants expressing $hpaG_{Xoo}$ from Xanthomonas oryzae pv. Oryzae, these results suggest that different harpins can affect the expression of different defense-related genes, as well as resistance to different plant pathogens.

• The Plant Pathology Journal
• /
• v.21 no.2
• /
• pp.127-131
• /
• 2005

In this comparative study, the effects of temperature, pH, and bactericides on the growth of Erwinia pyrifoliae and Erwinia amylovora were investigated. The maximum temperature for the growth of both Erwinia species was estimated to be $36{\circ}C$. The maximum specific growth rates of E. pyrifoliae and E. amylovora were observed at $27{\circ}C$ and $28{\circ}C$, respectively, and no significant growth differences were shown at their optimum temperatures. However, at lower temperatures ranging from 12-$21{\circ}C$, E. pyrifoliae showed higher growth rates with doubling times shorter than those of E. amylovora. Distinct growth rates at these temperatures revealed that E. pyrifoliae is more cold-tolerant than E. amylovora. The optimum pH for the growth of both pathogens was 7.5 and growth was not seen at pH ${\le}$ 5.0 and ${\ge}$ 10.0. These results showed that the effect of pH on the growth of E. pyrifoliae and E. amylovora was similar. Minimum inhibitory concentrations (MICs) of copper sulfate, oxolinic acid, streptomycin, and tetracycline, which inhibited growth of E. pyrifoliae and E. amylovora, were determined. The strains of both pathogens were able to grow at 0.08-0.32 mM copper sulfate, but not at higher concentrations. However, none of the tested strains grew in the presence of oxolinic acid (0.001 mM), streptomycin (0.1 mM), and tetracycline (0.01 mM) concentrations. These results suggested that all strains of both Erwinia species were sensitive to tested bactericides and indicated no occurrence of resistant strains of E. pyrifoliae in Korea.