Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot
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College of Plant Protection/Guangdong Province Key Laboratory of Microbial Signals and Disease Control,South China Agricultural University,Guangzhou 510642,China

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S432.1

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    Abstract:

    Banana bacterial sheath rot,caused by Dickeya dadantii,is a significant threat to the production of banana in China.Accurately and quantitatively detecting this pathogen at an early stage is important to control banana bacterial sheath rot.A SYBR GreenⅠreal-time quantitative PCR method was established with specific primers based on the housekeeping gene fusA of D.dadantii and optimized conditions for amplification.Comparisons were made between the result of the established SYBR GreenⅠreal-time quantitative PCR and that of the conventional PCR.The established SYBR GreenⅠreal-time quantitative PCR was used to detect XJ5-1 in banana leaf sheaths and soil samples inoculated with banana bacterial sheath rot bacteria XJ5-1.The results showed that the sensitivity of SYBR GreenⅠreal-time quantitative PCR for detecting the plasmid DNA was 3.3×10-5 ng/μL,100 times higher than that of conventional PCR.The established method was able to detect XJ5-1 in banana plants with a copy number of 4.31×103 copies/μL and in soil samples with a copy number of 1.07×109 copies/μL,and in samples of used greenhouse soil inoculated at an inoculum concentration of 1×103 CFU/mL.The sensitivity of detecting soil samples with the established method was 100 times higher than that with the conventional PCR.It is indicated that the established SYBR GreenⅠreal-time quantitative PCR method is rapid,simple,and sensitive.It will lay a foundation for the early diagnosis and monitoring of banana bacterial sheath rot.

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麦桂婉,肖文超,李云锋,李华平,饶雪琴. Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot[J]. Jorunal of Huazhong Agricultural University,2025,44(5):308-313.

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History
  • Received:December 25,2024
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  • Online: October 10,2025
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