[논문]Efficient Diagnosis of Ratoon Stunting Disease of Sugarcane by Quantitative PCR on Pooled Leaf Sheath Biopsies

Young, Anthony J.; Kawamata, Asuka; Ensbey, Mark A.; Lambley, Eleanore; Nock, Catherine J.

doi:10.1094/pdis-06-16-0848-re

Efficient Diagnosis of Ratoon Stunting Disease of Sugarcane by Quantitative PCR on Pooled Leaf Sheath Biopsies 원문보기

Plant disease, v.100 no.12, 2016년, pp.2492 - 2498

Young, Anthony J. (Centre for Crop Health, Faculty of Health, Engineering and Sciences, University of Southern Queensland, QLD, 4350, Australia) , Kawamata, Asuka (Southern Cross Plant Science, Southern Cross University, NSW, 2480, Australia) , Ensbey, Mark A. (Sunshine Sugar, Harwood Sugar Mill, Harwood Island, NSW, 2465, Australia) , Lambley, Eleanore (Southern Cross Plant Science, Southern Cross University) , Nock, Catherine J. (Southern Cross Plant Science, Southern Cross University)

Abstract ▼ AI-Helper

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 enzyme 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, whereas, 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 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 qPCR and PCR could detect L. xyli subsp. xyli in sugarcane ranging from 3 months to greater than 1 year old. Interpretive Summary Ratoon stunting disease (RSD) causes significant yield losses in sugarcane worldwide. However, there are no specific external symptoms that allow a farmer to determine which fields are infected; therefore, a range of diagnostic techniques have been developed. We report the results of a comparison between four different diagnostic methods and show that a highly efficient result can be achieved by using a DNA-based technique on lots of small samples taken from the cane. These samples are taken from the leaf sheath of the plant and, thus, are called leaf sheath biopsies (LSB). In fact, this works much better than previous methods where a small number of individual stalks were examined. Our results strongly suggest that RSD is more prevalent in Australia than has been generally acknowledged, and we believe that the use of the LSB sampling technique will help sugarcane farmers quickly identify infected crops and enjoy more sustainable and productive crops through better management of RSD.

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동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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Efficient Diagnosis of Ratoon Stunting Disease of Sugarcane by Quantitative PCR on Pooled Leaf Sheath Biopsies 원문보기

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