Recombinase Polymerase Amplification Assay for Rapid Field Diagnosis of Stewart’s Wilt of Corn Pathogen Pantoea stewartii subsp. stewartii

Lulu Cai, Qian Tian, Qingqing Meng, Xiaoyang Bao, Peidong Xu, Ji Liu, Wenjun Zhao () and Hui Wang ()
Additional contact information
Lulu Cai: Center for Biosafety, Chinese Academy of Inspection and Quarantine, Sanya 572024, China
Qian Tian: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China
Qingqing Meng: Center for Biosafety, Chinese Academy of Inspection and Quarantine, Sanya 572024, China
Xiaoyang Bao: Center for Biosafety, Chinese Academy of Inspection and Quarantine, Sanya 572024, China
Peidong Xu: Center for Biosafety, Chinese Academy of Inspection and Quarantine, Sanya 572024, China
Ji Liu: National Nanfan Research Institute (Sanya), Chinese Academy of Agricultural Sciences, Sanya 572024, China
Wenjun Zhao: Center for Biosafety, Chinese Academy of Inspection and Quarantine, Sanya 572024, China
Hui Wang: Southern Breeding Administrate Office of Hainan Province, Sanya 572000, China

Agriculture, 2023, vol. 13, issue 10, 1-14

Abstract: Stewart’s vascular wilt and leaf blight of sweet corn is caused by the Gram-negative enteric bacterium Pantoea stewartii subsp. stewartii . Stewart’s wilt results in substantial yield losses worldwide warranting rapid and accurate disease diagnosis. Recombinase polymerase amplification (RPA) is an isothermal technique that is tolerant to host plant-derived inhibitors and is, therefore, ideally suited for rapid in-field detection vis-à-vis traditional polymerase chain reaction-based molecular assays. An RPA assay coupled with a Lateral Flow Device (LFD) was developed for rapid, accurate, and sensitive real-time detection of P. stewartii subsp. stewartii directly from the infected host offering in-field pathogen detection, timely disease management, and satisfying quarantine and phytosanitary requirements. Twelve novel primer sets were designed against conserved genomic regions of P. stewartii subsp. Stewartii ; however, only the primers for amplification of the intergenic spacer region between capsular polysaccharide genes cpsA and cpsB were discernibly unique and adequate for unambiguous identification of P. stewartii subsp. stewartii . The P. stewartii subsp. stewartii -specific primers were further validated in a simplex RPA assay for specificity against twenty-six bacterial species representing several Pantoea and other closely related bacterial species/subspecies/strains found in the same niche, and naturally or artificially infected plant samples. The integrated RPA/LFD assay was also optimized for rapid and sensitive on-site detection of P. stewartii subsp. stewartii with an empirical detection limit of 0.0005 pg μL −1 bacterial DNA and 1 × 10 2 CFU mL −1 (app. two bacterial cells used per RPA reaction) in minimally processed samples for accurate, low-cost, and point-of-need diagnosis of the quarantine pathogen P. stewartii subsp. stewartii .

Keywords: corn leaf blight; Lateral Flow Device; Pantoea stewartii subsp. stewartii; phytosanitation; recombinase polymerase amplification; Stewart’s wilt (search for similar items in EconPapers)
JEL-codes: Q1 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 (search for similar items in EconPapers)
Date: 2023
References: View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.mdpi.com/2077-0472/13/10/1982/pdf (application/pdf)
https://www.mdpi.com/2077-0472/13/10/1982/ (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:gam:jagris:v:13:y:2023:i:10:p:1982-:d:1258546

Access Statistics for this article

Agriculture is currently edited by Ms. Leda Xuan

More articles in Agriculture from MDPI
Bibliographic data for series maintained by MDPI Indexing Manager ().