Multiplex PCR assay for the detection of Pantoea stewartii subsp. stewartii using species-specific genetic markers

被引：11

作者：

Thapa, Shree Prasad ^{[1
]}

Park, Duck Hwan ^{[2
]}

Wilson, Calum ^{[3
]}

Hur, Jang Hyun ^{[1
]}

Lim, Chun Keun ^{[2
]}

机构：

[1] Kangwon Natl Univ, Dept Environm Biol, Coll Agr & Life Sci, Chunchon 200701, South Korea

[2] Kangwon Natl Univ, Dept Appl Biol, Coll Agr & Life Sci, Chunchon 200701, South Korea

[3] Univ Tasmania, Sch Agr Sci, New Town Res Labs, Hobart, Tas 7001, Australia

来源：

AUSTRALASIAN PLANT PATHOLOGY | 2012年 / 41卷 / 05期

关键词：

Genetic marker; Multiplex PCR; P. stewartii subsp stewartii; Specific detection; POLYMERASE-CHAIN-REACTION; ERWINIA-STEWARTII; CLUSTER; WTSE;

D O I：

10.1007/s13313-012-0123-9

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

We developed a multiplex PCR detection assay for quick diagnosis of Pantoea stewartii subsp. stewartii the causal agent of Stewart's bacterial wilt and leaf blight of maize, using specific genetic markers. The genetic markers of wtsE, cpsA and hrpN gene specifically amplified 519 bp, 375 bp and 200 bp of DNA, respectively, from all isolates of P. stewartii subsp. stewartii but not from other phytopathogenic bacteria. The sensitivity of the genetic markers is down to 10(3) CFU/mL (5 cells per reaction) of bacterial culture and similar to 0.5 pg of genomic DNA. In sampling studies, the assay detected P. stewartii subsp. stewartii from artificially infected maize plants 12 days post inoculation. The PCR based genetic markers are specific and they give selective and sensitive amplification of P. stewartii subsp. stewartii for diagnosis in mixed cultures and in planta.

引用

页码：559 / 564

页数：6

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