National Key Laboratory of Green Pesticide, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Center for R&D of Fine Chemicals of Guizhou University, Guiyang 550025, China.
Int J Mol Sci. 2023 Jun 22;24(13):10515. doi: 10.3390/ijms241310515.
Citrus Huanglongbing (HLB) is one of the most destructive diseases in the citrus industry. At present, Liberibacter asiaticus (Las) cannot be cultured in vitro, and there is a lack of rapid methods to test antibacterial activity, which greatly hinders the discovery of new antibacterial agents against HLB. To establish a rapid screening method for antibacterial agents against HLB with simple operation, a short cycle, and a large number of tests, the Las contents in leaves from different citrus branches, different leaves from the same citrus branch, and two halves of the same citrus leaf were detected. Compared with the leaves on different branches and different leaves on the same branch, the difference in Las content of the left and right halves of the same leaf was small; the difference was basically between 0.7 and 1.3. A rapid and efficient method for primary screening agents against HLB termed the "half-leaf method" was established through our long-term optimization and improvement. To verify the stability and reliability of the activity data measured using this method, 6-chloropurine riboside, which is highly soluble in water, was used as the test agent, and its antibacterial activity against HLB was tested 45 times. The results of the antibacterial activity test showed little difference in the mean values of each data group, indicating that this method could be used as a rapid method for screening agents against HLB. We used this method to test the antibacterial activity of compounds synthesized by our research group against HLB and found that some of the compounds showed good activity.
柑橘黄龙病（HLB）是柑橘产业中最具破坏性的疾病之一。目前，亚洲韧皮杆菌（Las）不能在体外培养，缺乏快速检测抗菌活性的方法，这极大地阻碍了新的抗 HLB 抗菌剂的发现。为了建立一种操作简单、周期短、可大量检测的抗 HLB 抗菌剂快速筛选方法，检测了来自不同柑橘枝条的叶片、同一柑橘枝条上的不同叶片以及同一柑橘叶片两半的 Las 含量。与不同枝条上的叶片和同一枝条上的不同叶片相比，同一叶片左右两半的 Las 含量差异较小；差异基本在 0.7 到 1.3 之间。通过长期的优化和改进，建立了一种快速有效的抗 HLB 初级筛选剂的“半叶法”。为了验证该方法测量的活性数据的稳定性和可靠性，将高度水溶性的 6-氯嘌呤核苷用作测试剂，对其抗 HLB 活性进行了 45 次测试。抗菌活性测试的结果表明，每个数据组的平均值差异不大，表明该方法可作为筛选抗 HLB 剂的快速方法。我们使用该方法测试了我们研究小组合成的化合物对 HLB 的抗菌活性，发现一些化合物表现出良好的活性。
