文献类型： 外文期刊

作者： Fu, Shimin ¹ ; Wan, Hongcen ¹ ; Duan, Qiuyuan ¹ ; Wang, Xuefeng ¹ ; Lou, Binghai ² ; Bai, Ziqin ³ ; Zhou, Changyong ¹ ;

作者机构： 1.Southwest Univ, Citrus Res Inst, Integrat Sci Ctr Germplasm Creat Western China Sci, Natl Citrus Engn Res Ctr, Chongqing 400712, Peoples R China

2.Guangxi Acad Specialty Crops, Guangxi Key Lab Citrus Biol, Guilin 550006, Guangxi, Peoples R China

3.Guizhou Acad Agr Sci, Fruit Res Inst, Guiyang 550006, Guizhou, Peoples R China

关键词： Citrus Huanglongbing; CLas detection kit; Field application; NrdB

期刊名称：PHYTOPATHOLOGY RESEARCH （ 影响因子：3.5； 五年影响因子：3.9 ）

ISSN： 2096-5362

年卷期： 2025 年 7 卷 1 期

页码：

收录情况： SCI

摘要： Citrus Huanglongbing (HLB) is one of the most devastating diseases in the citrus industry and is caused primarily by 'Candidatus Liberibacter asiaticus' (CLas), a phloem-restricted gram-negative bacterium transmitted via citrus psyllids in the field. Precise CLas detection is crucial for HLB control, particularly during extensive surveys in new emerging regions. Unfortunately, the lack of a practical on-site detection method for CLas due to the limited specificity of immunostrips and the costive non-user friendliness approaches prevent the effective disease control. In this study, a probe-based recombinase polymerase amplification (RPA) targeting the CLas five-copy nrdB gene (beta-subunit of ribonucleotide reductase, RNR) was developed and evaluated for its specificity, sensitivity, and reliability. To enhance user-friendliness and facilitate widely use, an All-in-one kit was premade as freeze-dried pellets in individual tubes, containing all necessary components except DNA template. The applicability of the All-in-one kit for CLas detection was confirmed using plant or psyllid crude DNA extracts. Collectively, the All-in-one kit offers a specific, sensitive, rapid, cost-effective, and practical alternative for diagnosing CLas in field conditions when coupled with simplified crude DNA extraction method, providing an alternative approach for on-site CLas detection in the context of HLB.