The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean culti...
The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.
The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.
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제안 방법
2 kb avrXalO BamHI fragment in approximately 2,000 clones by colony hybridization. Based on the RFLP of cosmid DNA, a final six 이ones were elected and subcloned with pBluescript KS+ for DNA sequencing. Sequence analysis of the pXag30 clone revealed a single open reading frame (ORF) of AvrBs3-like protein in about 30 hypothetical proteins such as transposases, resolvases, and conjugal transfer proteins (Fig.
Electrophoretic separation and transfer to nylon membranes were performed as previously described [26]. DNA probes were labeled with α32P dCTP with the Megaprime DNA labeling kit (Amersham Biosciences, Piscataway, U.S.A.) and probed against DNA immobilized on a Hybond N+. Membrane hybridization and subsequent washing were carried out and then exposed to X-ray film or BAS film.
대상 데이터
2-kb BamHI fragment isolated from plasmid pBSavrXalO (data not shown). Based on the avrBs3 homologous band patterns with three different restriction enzymes EcoRI, BamHI, and Hindlll, the 155 bacterial strains isolated were divided into 6 groups, and OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 were selected as type strains (Fig. 2A). Differences among strains were obvious in the fragments between 3 and 4 kb (Fig.
glycines. SL1018 PM (3.7 kb plasmid-bome avr gene mutant) and SL1018 CM (5.2 kb chromosome-bome avr gene mutant) were selected among these mutants. According to the gene-for-gene theory, X.
Seeds of soybean (Glycine max L.) of American and Korean varieties were obtained from S. K. Lee (Seoul National University), and they were germinated on moist filter paper in Petri dishes for 2 and 3 days in the dark. Plants were grown in the greenhouse for evaluation of resistance at the seedling stage.
All strains ofX axonopodis pv. glycines used in this study were collected from farmers1 fields in Korea from 1999 to 2002. All strains collected were first tested for pathogenicity using nonhost plant tomato for HR and susceptible cultivar Pella.
성능/효과
In this study, the diversity of avirulence genes among Korean strains ofX axonopodis pv glycines, the bacterial leaf pustules pathogen, was revealed by avirulence gene homology. Korean strains ofX axonopodis pv lycines contained at least 3 copies of awBs3 gene family members, which were bom from plasmid or chromosomal DNA.
Based on the gene-for-gene hypothesis, a direct interaction occurs between a pathogen race-specificity gene (avr gene) product and the product of the complementary disease resistance gene (R gene) in the plant. The results of disease severity of representative strains on various soybean cultivars indicated that the difference of avrBs3 homolog patterns was related to the responses of soybean hosts that have the corresponding resistance gene.
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