Isolation and Characterization of a New Virulence Gene (abvA) of Agrobacterium tumefaciens
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概要
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A mutant (M-1) was isolated by transposon (Tn5) insertion mutagenesis of Agrobacterium tumefaciens (strain A-208, C58 chromosome, nopaline type T37 pTi, virulent). The M-1 mutant exhibited a complete avirulent phenotype on Kalanchoe daigremontiana leaf and Kalanchoe pinnata stem but a very attenuated virulent phenotype on root of Daucus carota. The mutant had one insertion of Tn5 in pTi. A wild-type target segment (2.3 kb) that included the site of Tn5 insertion in M-1 mutant was cloned. Introducing the 2.3 kb segment into M-1 complemented completely the avirulent phenotype, producing galls as big as strain A-208. The 2.3 kb segment was sequenced, identifying three open reading frames, ORF 1 (354 bp), ORF 2 (261 bp) and ORF 3 (801 bp) in the segment. A Tn5 was inserted between the third and fourth nucleotide of ORF 1 in M-1. The ORF i had no homology to any reported genes and thus was named the abvA gene. The ORF 3 had the high homology (identities 44%, positive 68%) to the gene of the safcosine oxidase β subunit (accession no. sp/P40875). Introduction of the DNA segment (743 bp) containing the abvA gene and its promoter region into M-1 partially complemented the avirulent phenotype of the mutant, producing galls smaller than strain A-208. The abvA gene was distributed not only on nopaline-type pTi (T37) but also on octopine-type pTi (A6NC) and chromosome (C58) of A. tume」aciens. M-1, being avirulent on K. daigremontiana and K. pinnata, had a Tn5 insertion only in the abvA gene on pTi but not in the abvA gene on the chromosome, implying that the abvA gene on the chromosome in strain A-208 is not functional. A binary vector, pIG121-Hm, containing the β-glucuronidase (GUS) gene with an intron was introduced into M-1, which was then applied to leaves of K. daigremontiana to assay GUS activity for monitoring T-DNA transfer to the host nucleus. High GUS activity comparable to that in strain A-208 was detected in M-1 in spite of its inability to induce galls, suggesting that M-1 can transfer T-DNA into the host nucleus, but cannot integrate it into the chromosome. (Received October 25, 2000 ; Accepted December 28, 2000)
- 日本植物病理学会の論文
- 2001-05-25
著者
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Nogawa Masahiro
Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
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Moriya Kei
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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KONDO Maki
Department of Cell Biology, National Institute for Basic Biology
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Majumder Parimal
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Kojima Mineo
Department of Applied Biology, Shinshu University
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Kojima M
Department Of Bioresource Chemistry Obihiro University Of Agriculture And Veterinary Medicine
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Nozue Masayuki
Department of Biology, Faculty of Science, Shinshu University
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SHIOIRI Hidenari
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
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Kojima Michiyuki
Department Of Bioresource Chemistry Obihiro University Of Agriculture And Veterinary Medicine
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Kojima Mineo
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Kondo Maki
Dept. Cell Biol. Nat. Inst. Basic Biol.
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Nozue Masayuki
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Kojima M
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Nogawa Masahiro
Division Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Nogawa Masahiro
Department Of Bioengineering Nagaoka University Of Technology
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Shioiri Hidenari
Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University
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Komai Michio
Nof Corporation
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Kojima Mineo
Department Of Applied Biological Science Faculty Of Textile Science And Technology Shinshu Universit
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