Detection of <EM>Camellia oleifera</EM> root rot pathogen with nested-PCR

LI He; SONG Guang-tao; HE Mo-jun; HAO Yan; ZHOU Guo-ying

Volume 26 Issue 6

Dec. 2009

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LI He, SONG Guang-tao, HE Mo-jun, HAO Yan, ZHOU Guo-ying. Detection of Camellia oleifera root rot pathogen with nested-PCR[J]. Journal of Zhejiang A&F University, 2009, 26(6): 849-853.

Citation:

LI He, SONG Guang-tao, HE Mo-jun, HAO Yan, ZHOU Guo-ying. Detection of Camellia oleifera root rot pathogen with nested-PCR[J]. Journal of Zhejiang A&F University, 2009, 26(6): 849-853.

Detection of Camellia oleifera root rot pathogen with nested-PCR

1.
College of Forestry，Central South University of Forestry & Technology，Changsha 410004，Hunan，China

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Corresponding author: ZHOU Guo-ying
Publish Date: 2009-12-20

Abstract

A species-specific polymerase chain reaction （PCR） assay for rapid and accurate detection of the pathogenic root rot of diseased Camellia oleifera plant tissues was developed. Based on differences in internal transcribed spacer（ITS） sequences of Fusarium proliferatum，a pair of species-specific primers，GF1/GR2，was designed. Other species of F. proliferatum were used to test the specificity of the primers. The GF1/GR2 primer only amplified a unique 400 bp band from the CSUFT070109 strain，and its detection sensitivity was 1 pg of genomic DNA in 25 μL reaction solution. A nested PCR procedure using ITS1/ITS4 as the first-round primers followed by GF1/GR2 increased detection sensitivity 10 000-fold to 100 ag. This assay detected the pathogen rapidly and accurately meaning methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.［Ch，4 fig. 22 ref.］
- forest protection,
- root rot of Camellia oleifera,
- Fusarium proliferatum,
- nested-PCR,
- molecular detection

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Detection of Camellia oleifera root rot pathogen with nested-PCR

1. College of Forestry，Central South University of Forestry & Technology，Changsha 410004，Hunan，China

Corresponding author: ZHOU Guo-ying

Publish Date: 2009-12-20

Key words:

Abstract: A species-specific polymerase chain reaction （PCR） assay for rapid and accurate detection of the pathogenic root rot of diseased Camellia oleifera plant tissues was developed. Based on differences in internal transcribed spacer（ITS） sequences of Fusarium proliferatum，a pair of species-specific primers，GF1/GR2，was designed. Other species of F. proliferatum were used to test the specificity of the primers. The GF1/GR2 primer only amplified a unique 400 bp band from the CSUFT070109 strain，and its detection sensitivity was 1 pg of genomic DNA in 25 μL reaction solution. A nested PCR procedure using ITS1/ITS4 as the first-round primers followed by GF1/GR2 increased detection sensitivity 10 000-fold to 100 ag. This assay detected the pathogen rapidly and accurately meaning methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.［Ch，4 fig. 22 ref.］

Detection of Camellia oleifera root rot pathogen with nested-PCR

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Detection of Camellia oleifera root rot pathogen with nested-PCR

1. College of Forestry，Central South University of Forestry & Technology，Changsha 410004，Hunan，China

Corresponding author: ZHOU Guo-ying

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Detection of Camellia oleifera root rot pathogen with nested-PCR

1. College of Forestry，Central South University of Forestry & Technology，Changsha 410004，Hunan，China

Corresponding author: ZHOU Guo-ying

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