文献类型： 外文期刊

作者： Li, Zhiting ¹ ; Feng, Wanzhen ¹ ; Zhu, Zaobing ³ ; Lu, Shengdan ¹ ; Lin, Mingze ¹ ; Dong, Jiali ⁴ ; Wang, Zhixin ¹ ; Liu, Fuxiu ⁵ ; Chen, Qinghe ¹ ;

作者机构： 1.Hainan Univ, Sch Breeding & Multiplicat, Sch Trop Agr & Forestry, Sanya, Peoples R China

2.Hainan Univ, Minist Educ, Key Lab Green Prevent & Control Trop Plant Dis & P, Haikou, Peoples R China

3.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing, Peoples R China

4.China Agr Univ, Sanya Inst, Sanya, Peoples R China

5.Postentry Quarantine Ctr Trop Plant, Haikou, Peoples R China

关键词： Phytophthora sojae; one-pot RPA/PCR; recombinase polymerase amplification; CRISPR/Cas12a; on-site detection; visual detection

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：4.0； 五年影响因子：5.1 ）

ISSN：

年卷期： 2024 年 15 卷

页码：

收录情况： SCI

摘要： Phytophthora sojae is a devastating plant pathogen that causes soybean Phytophthora root rot worldwide. Early on-site and accurate detection of the causal pathogen is critical for successful management. In this study, we have developed a novel and specific one-pot RPA/PCR-CRISPR/Cas12 assay for on-site detection (Cas-OPRAD) of Phytophthora root rot (P. sojae). Compared to the traditional RPA/PCR detection methods, the Cas-OPRAD assay has significant detection performance. The Cas-OPRAD platform has excellent specificity to distinguish 33 P. sojae from closely related oomycetes or fungal species. The PCR-Cas12a assay had a consistent detection limit of 100 pg. mu L-1, while the RPA-Cas12a assay achieved a detection limit of 10 pg. mu L-1. Furthermore, the Cas-OPRAD assay was equipped with a lateral flow assay for on-site diagnosis and enabled the visual detection of P. sojae on the infected field soybean samples. This assay provides a simple, efficient, rapid (<1 h), and visual detection platform for diagnosing Phytophthora root rot based on the one-pot CRISPR/Cas12a assay. Our work provides important methods for early and accurate on-site detection of Phytophthora root rot in the field or customs fields.