Tian Yanli, Zhao Yuqiang, Bai Sa, Walcott R R, Hu Baishi, Liu Fengquan
College of Plant Protection and Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, China.
Shanghai Agricultural Technology Extension and Service Center; Shanghai 201103, China.
Plant Dis. 2013 Jul;97(7):961-966. doi: 10.1094/PDIS-10-12-0930-RE.
A method was developed using a padlock probe (PLP) and dot-blot hybridization for detecting Acidovorax citrulli in cucurbit seed. The PLP was designed based on the 16S-23S internal transcribed spacer ribosomal DNA sequence from A. citrulli. The detection threshold for the PLP assay was 100 fg of genomic DNA, and A. citrulli was detected in 100% of artificially infested seedlots with 0.1% infestation or greater. In addition, using the PLP assay, 4 of 8 melon seedlots collected from Xinjang province and 15 of 47 watermelon seedlots collected from Ningxia province were positive for A. citrulli. In contrast, a conventional polymerase chain reaction (PCR) assay that relied on primers WFB1 and WFB2 facilitated A. citrulli detection in 1 of 8 and 5 of 47 seedlots from Xinjiang and Ningxia provinces, respectively. These data indicate that the PLP and dot-blot hybridization technique was more effective than conventional PCR for seed health testing.
开发了一种使用锁式探针（PLP）和斑点杂交检测葫芦科种子中西瓜嗜酸菌的方法。该PLP基于西瓜嗜酸菌的16S-23S内转录间隔区核糖体DNA序列设计。PLP检测方法的检测阈值为100 fg基因组DNA，在100%人工感染率为0.1%或更高的种子批次中检测到了西瓜嗜酸菌。此外，使用PLP检测方法，从新疆采集的8个甜瓜种子批次中有4个、从宁夏采集的47个西瓜种子批次中有15个对西瓜嗜酸菌呈阳性。相比之下，一种依赖引物WFB1和WFB2的传统聚合酶链反应（PCR）检测方法分别在来自新疆和宁夏的8个种子批次中的1个以及47个种子批次中的5个中检测到了西瓜嗜酸菌。这些数据表明，PLP和斑点杂交技术在种子健康检测方面比传统PCR更有效。
