Efficient diagnosis of ratoon stunting disease of sugarcane by quantitative PCR on pooled leaf sheath biopsies

Young, Anthony J. and Kawamata, Asuka and Ensbey, Mark A. and Lambley, Eleanore and Nock, Catherine J. (2016) Efficient diagnosis of ratoon stunting disease of sugarcane by quantitative PCR on pooled leaf sheath biopsies. Plant Disease, 100 (12). pp. 2492-2498. ISSN 0191-2917

Abstract

Ratoon stunting disease (RSD), caused by the bacterium Leifsonia xyli subsp. xyli, is arguably one of the most devastating diseases of sugarcane. Four diagnostic techniques were compared for 100 fields of sugarcane (Saccharum interspecific hybrids) of unknown infection status. These were quantitative polymerase chain reaction on pooled leaf sheath biopsies (LSB-qPCR), conventional PCR on the same templates (LSB-PCR), evaporative-binding immunoassay (EB-EIA) coupled with phase contrast microscopy (PCM) on expressed xylem sap from the same fields, and conventional PCR on the same xylem sap samples. LSB-qPCR and LSB-PCR detected the causal agent in 27 and 18 fields respectively, while from samples of expressed xylem sap from the same fields, conventional PCR identified 12 infections, and EB-EIA/PCM detected L. xyli subsp. xyli in 3 fields. The sensitivities of qPCR and PCR were approximately 103 CFU ml-1 and 104 CFU ml-1 respectively, determined from plate counts of a dilution series. Tests were conducted on a further 139 LSB samples from across the Australian industry, with qPCR and PCR diagnosing RSD in 31 and 25 fields respectively. Using qPCR and PCR on LSB samples, RSD was diagnosed in a range of cultivars throughout the year, and could detect L. xyli subsp. xyli in sugarcane ranging from 3 months to greater than 1 year old.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Files associated with this item cannot be displayed due to copyright restrictions.
Faculty / Department / School: Current - Faculty of Health, Engineering and Sciences - School of Agricultural, Computational and Environmental Sciences
Date Deposited: 24 Feb 2017 04:01
Last Modified: 27 Sep 2017 03:42
Uncontrolled Keywords: ratoon stunting disease (RSD)
Fields of Research : 07 Agricultural and Veterinary Sciences > 0703 Crop and Pasture Production > 070308 Crop and Pasture Protection (Pests, Diseases and Weeds)
Socio-Economic Objective: D Environment > 96 Environment > 9604 Control of Pests, Diseases and Exotic Species > 960413 Control of Plant Pests, Diseases and Exotic Species in Farmland, Arable Cropland and Permanent Cropland Environments
Identification Number or DOI: 10.1094/PDIS-06-16-0848-RE
URI: http://eprints.usq.edu.au/id/eprint/30683

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