Establishment of a LAMP Assay for Rapid Detecting <i>Diaporthe citri </i>on Pomelo

LAI Baochun; YAO Jinai

doi:10.19303/j.issn.1008-0384.2022.011.013

Volume 37 Issue 11

Nov. 2022

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Fujian Journal of Agricultural Sciences > 2022 > 37(11): 1470-1475. > DOI: 10.19303/j.issn.1008-0384.2022.011.013

LAI B C, YAO J N. Establishment of a LAMP Assay for Rapid Detecting Diaporthe citri on Pomelo [J]. Fujian Journal of Agricultural Sciences，2022，37（11）：1470−1475. DOI: 10.19303/j.issn.1008-0384.2022.011.013

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Establishment of a LAMP Assay for Rapid Detecting Diaporthe citri on Pomelo

LAI Baochun^1,,
YAO Jinai^2, ,

1.
Zhangzhou Institute of Agricultural Sciences, Zhangzhou, Fujian　363005, China
2.
Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests/Fujian Engineering Research Center for Green Pest Management, Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China

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Received Date: August 31, 2022
Revised Date: October 09, 2022
Available Online: December 27, 2022

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Abstract

Abstract

Objective A loop-mediated isothermal amplification (LAMP) method for visual, rapid, and accurate detection of Diaporthe citri on pomelos in early stage of black spot disease was developed.
Method Based on the elongation factor-1α (EF-1α) sequence of D. citri, a set of LAMP primers was designed. Template DNA from the infected leaves was used to establish the optimal temperature and time for the LAMP operation. Assay specificity and sensitivity were verified by positive detection of the infected plants in the field.
Result The newly established LAMP method could effectively and specifically detect D. citri on pomelo at the optimal temperature of 65 ℃ with 60 m for the reaction time. The detection limit of the assay on D. citri was 10 fg·μL⁻¹. On 15 field samples with the typical black spot symptoms, a 100% positive detection rate was achieved by the LAMP assay.
Conclusion The established visual and rapid LAMP assay demonstrated a high specificity, sensitivity, and perfect detection of D. citri on diseased pomelo. It was considered appropriate for field application.
- Pomelo,
- black spot,
- Diaporthe citri,
- loop-mediated isothermal amplification,
- detection

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References

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