Rapid differentiation of Fusarium oxysporum isolates using PCR-SSCP with the combination of pH-variable electrophoretic medium and low temperature

被引：4

作者：

Dong, DX ^{[1
]}

Liu, HM ^{[1
]}

Peng, HS ^{[1
]}

Huang, XQ ^{[1
]}

Zhang, XH ^{[1
]}

Xu, YQ ^{[1
]}

机构：

[1] Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Mol Microbiol Lab, Shanghai 200240, Peoples R China

来源：

ELECTROPHORESIS | 2005年 / 26卷 / 22期

关键词：

differentiation of Fusarium oxysporum; low temperature; pH-variable electrophoretic medium; SSCP;

D O I：

10.1002/elps.200500315

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Differentiation of Fusarium oxysporum is significantly important for unraveling the pathogenetic mechanism of Fusaria wilts. In this study, isolates of F oxysporum were screened from the soils in the rhizosphere of watermelon plant by Komada medium and differentiated by SSCP approach with the combination of pH-variable electrophoretic medium (Tris-MES-EDTA (TME), pH 6.1) and low temperature (9 degrees C). We found that TIME was a good electrophoretic medium and its pH value was variable over the course of electrophoresis in our apparatus. The pH-variable electrophoretic medium made more contribution for the better differentiation of F oxysporum isolates than low temperature. The combination of TME pH 6.1 and low temperature showed an improved effect on resolution of ssDNAs. Leaving partial nondenatured dsDNA for SSCP was advantageous for differentiation of F oxysporum isolates. The SSCP patterns of F oxysporum isolates proved to be highly reproducible. Sequencing data confirmed that this SSCP method could detect one single base change within the 550 bp PCR fragment from the ribosomal internal transcribed spacer region of F oxysporum.

引用

页码：4287 / 4295

页数：9

共 21 条

[1]

Alves-Santos FM, 1999, APPL ENVIRON MICROB, V65, P3335

[2]

[Anonymous], 1990, PCR PROTOCOLS, DOI DOI 10.1016/B978-0-12-372180-8.50042-1

[3] Gene genealogies and AFLP analyses in the Fusarium oxysporum complex identify monophyletic and nonmonophyletic formae speciales causing wilt and rot disease [J].