Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting <italic style="font-style: italic">Dickeya dadantii</italic> of banana bacterial sheath rot

MAI Guiwan; XIAO Wenchao; LI Yunfeng; LI Huaping; RAO Xueqin

doi:10.13300/j.cnki.hnlkxb.2025.05.031

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Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot

Techniques and methods | 更新时间：2026-11-05

- Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot
- Journal of Huazhong Agricultural University Vol. 44, Issue 5, Pages: 308-313(2025)
- 作者机构：
  
  华南农业大学植物保护学院/广东省微生物信号与病害防治重点实验室，广州510642
- 作者简介：
- 基金信息：
- DOI：10.13300/j.cnki.hnlkxb.2025.05.031
  CLC： S432.1
- Received：25 December 2024，
  
  Published：15 September 2025
- 稿件说明：
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麦桂婉，肖文超，李云锋，等.香蕉细菌性鞘腐病菌实时荧光定量PCR检测方法的建立［J］.华中农业大学学报，2025，44（5）：308‐313.

MAI Guiwan,XIAO Wenchao,LI Yunfeng,et al.Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot[J].Journal of Huazhong Agricultural University,2025,44(05):308-313.
麦桂婉，肖文超，李云锋，等.香蕉细菌性鞘腐病菌实时荧光定量PCR检测方法的建立［J］.华中农业大学学报，2025，44（5）：308‐313. DOI： 10.13300/j.cnki.hnlkxb.2025.05.031.

MAI Guiwan,XIAO Wenchao,LI Yunfeng,et al.Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot[J].Journal of Huazhong Agricultural University,2025,44(05):308-313. DOI： 10.13300/j.cnki.hnlkxb.2025.05.031.

摘要

为实现香蕉细菌性鞘腐病的早期准确检测，基于香蕉细菌性鞘腐病菌的管家基因

fusA

设计特异性引物，优化扩增条件，建立了SYBR GreenⅠ实时荧光定量PCR检测方法，并与常规PCR进行比较。结果显示，SYBR GreenⅠ实时荧光定量PCR检测质粒DNA的灵敏度为3.3×10

-5

ng/µL，是常规PCR的100倍。利用建立的方法对接种香蕉细菌性鞘腐病菌XJ5-1的香蕉叶鞘和土样进行检测，结果发现该方法能检测到香蕉叶鞘中拷贝数为4.31×10

copies/µL和土样中拷贝数为1.07×10

copies/µL的XJ5-1；能检测出接种浓度为1×10

CFU/mL的温室土样中的XJ5-1，灵敏度是常规PCR的100倍。结果表明，建立的SYBR GreenⅠ实时荧光定量PCR法快速简便、灵敏度高，可实现香蕉细菌性鞘腐病的早期诊断。

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

D.dadantii

and optimized conditions for amplification.Comparisons were made between the result of the established SYBR GreenⅠreal-time qu

antitative 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 4.31×10

copies/μL and in soil samples with a copy number of 1.07×10

copies/μL，and in samples of used greenhouse soil inoculated at an inoculum concentration of 1×10

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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references

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Establishment of SYBR GreenⅠreal-time quantitative PCR for detecting Dickeya dadantii of banana bacterial sheath rot

DOI：10.13300/j.cnki.hnlkxb.2025.05.031

摘要

Abstract

关键词

Keywords

references