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Occurrence of the strA-strB Streptomycin Resistance Genes in Pseudomonas Species Isolated from Kiwifruit Plants 원문보기

The journal of microbiology, v.42 no.4, 2004년, pp.365 - 368  

Han Hyo Shim (Department of Biology, Sunchon National University) ,  Koh Young Jin (Department of Applied Biology, Sunchon National University) ,  Hur Jae-Seoun (Department of Environmental Education, Sunchon National University) ,  Jung Jae Sung (Department of Biology, Sunchon National University)

Abstract ▼ AI-Helper 아이콘AI-Helper

The occurrence of strA-strB streptomycin-resistance genes within transposon Tn5393 was examined in Pseudomonas syringae pv. actinidiae, P. syringae pv. syringae, and P. marginalis, isolated from kiwifruit plants in Korea and Japan. PCR amplification with primers specific to strA-strB revealed that t...

주제어

#Pseudomonas syringae pv   #syringae   #kiwifruit   #strA-strB   #Tn5393  

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제안 방법

  • In order to demonstrate that our three Pseudomonas species contained strA-strB in Tn5393, PCR experiments were performed with primers designed to amplify a 1, 640-bp region of strA-strB, a 300-bp region of tnpA, and a 949-bp region which included res and tnpR. The PCR products amplified with the three Pseudomonas species were identical to the size expected for Tn5393 (Fig.
  • The presence of strA-strB genes was confirmed by primers designed from the 5* end of strA, and from the 3' end of strB: strAB-1, 5'-TTGAATCGAACTAATAT-3'; strAB-2, 5'・CTAGTATGACGTCTGTCG-3L For the specific detection of the tnpA sequence, we used the tnpA primers: tnpA-F, 5'-GGCGGGATCTGCTTGTAGAG-3' corresponding to tnpA positions 1, 193 to 1, 212, and tnpA-R, 5'-CTC- CGGAGATGTCTGGCTTACT-3', corresponding to tnpA positions 1, 471 to 1, 492. In order to detect IS elements which might be located upstream ofstrA-strB, a PCR primer set was designed from tnpA and str A: IS-F, 5'-ATGC- CCAAGGTCTTCATCAC-3, , corresponding to tnpA positions 2, 773 to 2, 792, and IS-R, 5'-TCACCACGTCGA- AAAAC AAA-3' corresponding to str A positions 65 to 84. Three Pseudomonas species were cultured for 24 h on peptone-sucrose broth at 30C, and whole cell DNA was prepared, as previously described (Hanef al.
  • Streptomycin has been used extensively to control this disease in both Korea and Japan. In this work, we assessed the occurrence of the Tn5595-like transposon, and analyzed the nucleotide polymorphisms of the strA-strB genes within three streptomycin-resistant Pseudomonas species, all of which were isolated from kiwifruit plants in both countries.

이론/모형

  • The annealing temperatures were 48℃ for the amplification of the strA-strB and tnpA sequences, and 55℃ for the detection of IS elements. The nucleotide sequence analysis was conducted at the Macrogen Co. (Korea) using the doublestranded dideoxy sequencing method. The Basic Local Alignment Search Tool (BLAST) search system was used for sequence comparisons.
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참고문헌 (11)

  1. Balestra, G.M. and L. Varvaro. 1997. Pseudomonas syringae pv. syringae causative agent of disease on floral buds of Actinidia deliciosa (A. Chev) Liang et Ferguson in Italy. J. Phytopathology 145, 375-378 

  2. Burr, T.J., J.L. Norelli, and C.L. Reid. 1993. Streptomycin-resistant bacteria associated with fire blight infections. Plant Dis. 77, 63-66 

  3. Chiou, C.-S. and A.L. Jones. 1993. Nucleotide sequence analysis of a transposon (Tn5393) carrying streptomycin resistance genes in Erwinia amylovora and other Gram-negative bacteria. J. Bacteriol. 175, 732-740 

  4. Han, H.S., H.Y. Nam, Y.J. Koh, J.-S. Hur, and J.S. Jung. 2003a. Molecular bases of high-level streptomycin resistance in Pseudomonas marginalis and Pseudomonas syringae pv. actinidiae. J. Microbiol. 41, 16-21 

  5. Han, H.S., Y.J. Koh, J.-S. Hur, and J.S. Jung. 2003b. Identification and characterization of coronatine-producing Pseudomonas syringae pv. actinidiae. J. Microbiol. Biotechnol. 13, 110-118 

  6. LAbee-Lund, T.M. and H. Sørum. 2000. Functional Tn5393-like transposon in the R plasmid pRAS2 from the fish pathogen Aeromonas salmonicida subspecies salmonicida isolated in Norway. Appl. Environ. Microbiol. 66, 5533-5535 

  7. Lioa, C.H., D.E. McCallus, W.F. Fett, and Y. Yang. 1997. Identification of gene loci controlling pectate lyase production and soft-rot pathogenicity in Pseudomonas marginalis. Can. J. Microbiol. 43, 425-431 

  8. Shaw, K.J., P.N. Rather, R.S. Hare, and G.H. Miller. 1993. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol. Rev. 57, 138-163 

  9. Sundin, G.W. 2002. Distinct recent lineages of the strA-strB streptomycin-resistance genes in clinical and environmental bacteria. Curr. Microbiol. 45, 63-69 

  10. Sundin, G.W. and C.L. Bender. 1996. Dissemination of the strAstrB streptomycin-resistance genes among commensal and pathogenic bacteria from humans, animals, and plants. Mol. Ecol. 5,133-143 

  11. Sundin, G.W. and C.L. Bender. 1995. Expression of the strA-strB streptomycin resistance gene in Pseudomonas syringae and Xanthomonas campestris and characterization of IS6100 in X. campestris. Appl. Environ. Microbiol. 61, 2891-2897 

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