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A nalysis ofphcA between different pathogenicity of Ralstonia solanacearum

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Materials and methods Strains FJAT-91 （ V ）、 FJAT-98 （ A ） and GMI1000

Cloning of phcA （ Genin et al. 2005 ）

A 1.3 kb DNA fragment encompassing the phcA

open reading frame was PCR-amplified using the

primers phc1 and pCA2

Sequence Analysis of phcA

TA Cloning and Sequencing （ BioSune ）

Data Base:NCBI 、 Uniprot and Nr

A nalysis ofphcA between different pathogenicity of Ralstonia solanacearum

Results and analysis

Cloning of phcA—3000

—1000

—500

M: DNA marker; "+": GMI1000; "-":ddw; 1: FJAT-91; 2: FJAT-98

Fig. 1 PCR-amplified of phcA of FJAT-91 and FJAT-98

A nalysis ofphcA between different pathogenicity of Ralstonia solanacearum

Results and analysis TA Cloning and analysis of IS:2032

A nalysis ofphcA between different pathogenicity of Ralstonia solanacearum

Fig. 2 Sequence of FJAT-98

IS ： 2032bp

virulence

virulence

gene 1 ：

gene 2 ：

gene 3 ：

IS:

Fig. 3 Analysis of IS sequence

Gene 1:transferase(tnpA); Gene 2:Methyltransferases;

Gene 3:Unkown protein

Materials and methods Strains Ralstonia solanacearum from different host plants （ 84 strains ）

BOX PCR-amplified and REP PCR-amplified

84 isolates Ralstonia solanacearum from different host plants were subjected to genomic fingerprinting using primer sets corresponding to BOX and REP elements （ Versalovic et al.,1994 ）

Cluster analysis of the BOX-PCR and REP-PCR DNA fingerprints

Cluster analyses in SPSS were conducted with METHOD= Between-groups linkage and Furthest neighbor and Ward's method

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Results and analysis BOX PCR-amplified

Fig. 4 BOX-PCR amplified of Ralstonia solanacearum from different host plants

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Results and analysis REP PCR-amplified

Fig. 5 REP-PCR amplified of Ralstonia solanacearum from different host plants

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Results and analysis

Cluster analysis of the BOX-PCR and REP-PCR DNA fingerprints

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

λ=22

Cluster of BOX-PCR and REP-PCR analysis showed that genetic diversity of R. solanacearum was concerned with geographic origin and host plants, and different host

plants played a leading role in the genetic difference of R. solanacearum.

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Results and analysis

Cluster analysis of the BOX-PCR DNA fingerprints

λ=20

BOX-PCR was useful in separating strains based on geographic origin.

Results and analysis

Cluster analysis of the REP-PCR DNA fingerprints

λ=8

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

REP-PCR was useful in separating strains based on host plant.

Genetic diversity analysis of Ralstonia solanacearum based on BOX-PCR and REP-PCR

Results and analysis Different host plants played a leading role in the genetic

difference of R. solanacearum. BOX-PCR was useful in separating strains based on

geographic origin. 700 bp and 900 bp was amplified in MinDong,200 bp 、 750

bp and 950 bp was amplified in MinBei,700 bp and 2000 bp was amplified in MinZhong.

REP-PCR was useful in separating strains based on host plant.

1050 bp and 2000 bp was amplified in eggplant and parts of tomato,1700 bp and 2100 bp was amplified in chilli,1250 bp 、 1500 bp and 3000 bp was not amplified in the parts of chilli and tomato.

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