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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Microbe and Virus Interactions with Plants
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1537812
This article is part of the Research Topic Diagnostic Innovations for Microbial Pathogens in Edible Plants: Cutting-Edge Technologies for Enhanced Detection View all 3 articles

Development and validation of a LAMP-based method for rapid and reliable detection of Xanthomonas albilineans, the causal agent of sugarcane leaf scald

Provisionally accepted
Moutoshi Chakraborty Moutoshi Chakraborty 1,2*Shamsul Bhuiyan Shamsul Bhuiyan 3,4Simon Strachan Simon Strachan 1,4Muhammad J A Shiddiky Muhammad J A Shiddiky 5Nam-Trung Nguyen Nam-Trung Nguyen 4Rebecca Ford Rebecca Ford 1,2
  • 1 School of Environment and Science, Griffith Sciences, Griffith University, Nathan, Australia
  • 2 Centre for Planetary Health and Food Security, Griffith University, Nathan, Queensland, Australia
  • 3 Sugar Research Australia (Australia), Brisbane, Queensland, Australia
  • 4 Queensland Micro and Nanotechnology Centre, Griffith University, Brisbane, Queensland, Australia
  • 5 Charles Sturt University, Bathurst, New South Wales, Australia

The final, formatted version of the article will be published soon.

    Xanthomonas albilineans (Xalb)-induced leaf scald (LS) is a significant bacterial disease affecting sugarcane and posing a global threat to the sugarcane industry. The presence of irregular symptoms makes traditional phenotypic detection difficult, and molecular methods necessitate costly equipment, labor, and extended sample-to-answer processing times. In this study, we introduce an innovative 'rapid' DNA isolation method that requires no reagents, along with an isothermal amplification-based assay for efficient detection of Xalb DNA in sugarcane xylem sap, leaf tissue, and meristematic tissue samples. The approach involves subjecting sugarcane samples obtained from infected plants to heat lysis, followed by loop-mediated isothermal amplification (LAMP)-based fluorescence and colorimetric quantification of Xalb within a single microcentrifuge tube. This method exhibited exceptional detection sensitivity (detecting as low as 1 cell/µL), reproducibility (with a standard deviation (SD) of <5% for n = 3), and a broad linear dynamic range (ranging from 10 pM to 1 aM or 10 7 -10 0 copies/µL, r = 0.99). Moreover, the quantification of Xalb was accurately correlated with sugarcane cultivar disease ratings. The results were validated using q-PCR, showing 91-98% agreement. This assay is also showed effectiveness in determining the optimal sampling times and plant parts by monitoring the progression of the disease over time. This diagnostic assay holds significant potential as a commercial opportunity for a kit-based DNA extraction/purification-free molecular detection alternative that may adapted into a handheld device, enabling on-farm detection and quantification of the pathogen responsible for LS disease.

    Keywords: Nucleic acid isolation, plant pathogen diagnostic, LS disease, Xanthomonas albilineans, Rapid detection

    Received: 01 Dec 2024; Accepted: 14 Jan 2025.

    Copyright: © 2025 Chakraborty, Bhuiyan, Strachan, Shiddiky, Nguyen and Ford. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Moutoshi Chakraborty, School of Environment and Science, Griffith Sciences, Griffith University, Nathan, Australia

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.