文献类型：期刊文献

英文题名：Development of a new ERA-CRISPR/Cas12a method for rapid sensitive detection of Ralstonia pseudosolanacearum in eucalyptus

作者：Peng, Xiaoping[1,2] Wang, Shengkun[2] Zhang, Yipin[1] Wang, Shengjie[2] Meng, Sen[2] Hu, Lipan[1] Ma, Haibin[2]

第一作者：Peng, Xiaoping

通信作者：Hu, LP[1];Ma, HB[2]

机构：[1]Chongqing Three Gorges Univ, Coll Biol & Food Engn, Chongqing, Peoples R China;[2]Chinese Acad Forestry, Res Inst Trop Forestry, State Key Lab Tree Genet & Breeding, Guangzhou, Peoples R China

年份：2025

卷号：16

外文期刊名：FRONTIERS IN PLANT SCIENCE

收录：;Scopus(收录号：2-s2.0-105024464503);WOS:【SCI-EXPANDED(收录号:WOS:001632975000001)】；

基金：The author(s) declare financial support was received for the research and/or publication of this article. This work was supported by the National Key R&D Program of China (2023YFC2604800) and the Natural Science Foundation of Chongqing, China (CSTB2023NSCQ-MSX0021).

语种：英文

外文关键词：Ralstonia pseudosolanacearum; ERA-CRISPR/Cas12a; eucalyptus; rapid detection; visualization

摘要：Introduction Ralstonia pseudosolanacearum is a significant pathogenic bacterium that causes bacterial wilt in Eucalyptus worldwide. Asymptomatic Eucalyptus cuttings may harbor substantial quantities of R. pseudosolanacearum, leading to latent infections that increase the risk of pathogen dissemination. Currently, there are no effective methods available to cure Eucalyptus bacterial wilt; therefore, rapid and sensitive detection methods for this disease are urgently needed to mitigate losses in the Eucalyptus industry.Methods In this study, we developed a rapid and accurate diagnostic method for detecting R. pseudosolanacearum based on enzymatic recombinase amplification (ERA) combined with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a technology.Results The ERA-CRISPR/Cas12a method demonstrated high specificity and exhibited no cross-reactivity with other common bacterial pathogens. The detection limit for R. pseudosolanacearum by the fluorescence and the LFS detection system was as low as 100 copies/mu L. Furthermore, the results can be visualized through an ERA-CRISPR/Cas12a fluorescent signal (ERA-CRISPR/Cas12a-FL), color under blue light or an ERA-CRISPR/Cas12a lateral flow strip (ERA-CRISPR/Cas12a-LFS).Discussion The newly developed ERA-CRISPR/Cas12a method could detect R. pseudosolanacearum in Eucalyptus rapidly and accurately. Moreover, the samples can be detected within one hour by our developed method, highlighting the significant potential for onsite applications in disease management.