Shu Ran, Yin Xianhui, Long Youhua, Yuan Jun, Zhou Houyin
Engineering and Technology Research Center of Kiwifruit, Guizhou University, Guiyang, China.
Institute of Crop Protection, Guizhou University, Guiyang, China.
Front Microbiol. 2022 May 25;13:896567. doi: 10.3389/fmicb.2022.896567. eCollection 2022.
Plum bacterial shot-hole caused by (. ) is one of the primary bacterial diseases in plum tree planting areas, resulting in abnormal growth of plum trees and severe economic losses. Early diagnosis of . is crucial to effectively control plant diseases. In this study, loop-mediated isothermal amplification (LAMP) analysis for genome-specific gene sequences was developed for the specific detection of . . We designed the LAMP primers based on the gene of . The best reaction system was 0.2 μmol·L for outer primer F3/B3 and 1.6 μmol·L for inner primer FIP/BIP. The LAMP reaction was optimal at 65°C for 60 min based on the color change and gel electrophoresis. This technology distinguished from other control bacteria. The detection limit of the LAMP technology was 5 fg·μl genomic DNA of . , which is 1,000 times that of the traditional PCR detection method. The LAMP technology could effectively detect the DNA of from the infected leaves without symptoms after indoor inoculation. Furthermore, the LAMP technology was applied successfully to detect field samples, and the field control effect of 0.3% tetramycin after LAMP detection reached 82.51%, which was 7.90% higher than that of conventional control. The proposed LAMP detection technology in this study offers the advantages of ease of operation, visibility of results, rapidity, accuracy, and high sensitivity, making it suitable for the early diagnosis of plum bacteria shot-hole disease.
由（. ）引起的李细菌性穿孔病是李树种植区的主要细菌性病害之一，导致李树生长异常并造成严重经济损失。对（. ）进行早期诊断对于有效控制植物病害至关重要。在本研究中，开发了基于基因组特异性基因序列的环介导等温扩增（LAMP）分析方法用于（. ）的特异性检测。我们基于（. ）的（基因名称未给出）基因设计了LAMP引物。最佳反应体系为外引物F3/B3浓度0.2 μmol·L，内引物FIP/BIP浓度1.6 μmol·L。基于颜色变化和凝胶电泳，LAMP反应在65°C下60分钟最为适宜。该技术能够将（. ）与其他对照细菌区分开来。LAMP技术的检测限为（. ）基因组DNA 5 fg·μl，是传统PCR检测方法的1000倍。LAMP技术能够有效检测室内接种后无症状感染叶片中的（. ）DNA。此外，LAMP技术成功应用于田间样本检测，LAMP检测后0.3%四霉素的田间防治效果达到82.51%，比常规防治高7.90%。本研究中提出的LAMP检测技术具有操作简便、结果可视化、快速、准确和高灵敏度等优点，适用于李细菌性穿孔病的早期诊断。
