Establishment of a recombinase polymerase amplification detection method for Puccinia striiformis f. sp. tritici.
Establishment of a recombinase polymerase amplification detection method for Puccinia striiformis f. sp. tritici.
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DOI：
10.1038/s41598-023-42663-4
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发表时间：
2023-09-26
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
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作者：
小麦条锈病是由条锈菌（Pucciniastriiformis f.）小麦锈病（Pst）是一种气传病害，在小麦的整个生长期内危害小麦。本研究以Pst的Pst134EA_003354未知蛋白（GenBank：XM_047941824.1）为靶序列，设计引物PS-RPA-F和PS-RPA-R，以及探针PS-LF-probe，进行重组酶聚合酶扩增（RPA）技术。将流动色谱法与该工艺相结合，建立了Pst的RPA检测方法。该方法在39 ℃恒温条件下成功建立了10 min内的目视检测，检测结果与普通PCR分析结果一致。但该方法仅对Pst有较高的特异性，检测限为10 fg/μL。此外，这种快速的方法成功地检测到Pst从小麦叶片在田间培养期间，表明应用的实际效益。综上所述，本研究建立的RPA检测方法具有高效、功能简单、快速、普遍实用的特点，为Pst的早期检测和预防提供了理论依据。
Wheat stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is an airborne disease that endangers wheat during its entire growth period. In this study, the Pst134EA_003354 uncharacterized protein (GenBank: XM_047941824.1) of Pst was used as the target sequence, and the primers PS-RPA-F and PS-RPA-R, as well as the probe PS-LF-probe, were designed for recombinase polymerase amplification (RPA) technology. Flow chromatography was combined with the process to establish an RPA detection method for Pst. This method successfully established visual detection within 10 min under a constant temperature of 39 °C, and the detection results were consistent with those of ordinary PCR analysis. However, it only had high specificity for Pst, and the detection limit was 10 fg/μL. In addition, this rapid method successfully detected Pst from wheat leaves during the field incubation period, indicating substantial benefits for applied use. In summary, the RPA detection method established in this study has the favourable characteristics of high efficiency, simple functionality, and rapid and universal practicability, providing a theoretical basis for the early detection and prevention of Pst.
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影响因子：
5.7
作者：
通讯作者：
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影响因子：
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作者：
Cao, Yuhao;Yan, Dankan;Song, Xuemei
通讯作者：
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影响因子：
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作者：
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影响因子：
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作者：
Li JJ;Xiong C;Liu Y;Liang JS;Zhou XW
通讯作者：
Zhou XW
DOI：
10.13926/j.cnki.apps.000342
发表时间：
2020-01-01
期刊：
Acta Phytopathologica Sinica
影响因子：
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作者：
Ju, Yu-Liang;Shen, Peng-Fei;Pan, Y. M.
通讯作者：
Pan, Y. M.