Abstract: Gray mold caused by Botrytis cinerea is one of the main diseases of tomatoes，which severely affects the yield and quality of tomatoes. In order to achieve the early and rapid detection of tomato gray mold，by using the ACTIN gene of Botrytis cinerea as the target gene，and based on the loop‐mediated isothermal amplification（LAMP）technology，a set of LAMP specific primers was designed and screened，and the reaction system and reaction conditions were optimized to realize the rapid isothermal amplification of Botrytis cinerea.Through agarose gel electrophoresis and SYBR Green Ⅰ visualization analysis，the optimal dosages of Bst Ⅱ DNA polymerase and dNTPs，as well as the optimal ratio of inner and outer primers，were determined to be 0.6 U/μL，1.25 mmol/L，and 2∶1 respectively.The specific detection of Botrytis cinerea could be achieved at 61 ℃ for 40 min，and its sensitivity could reach 100 ag/μL，which was 106 times the sensitivity of ordinary PCR detection. When this method was applied to the detection of tomato diseases，the spores detection limit for Botrytis cinerea could reach 20 spores/mL，and the pathogen could be detected in tomato leaves that had been infected for 4 days without obvious phenotypic symptoms of gray mold，making it applicable for the early，rapid，sensitive and visual detection of tomato gray mold.

Key words: Tomato, Botrytis cinerea, Gray mold, Loop‐mediated isothermal amplification, ACTIN gene

摘要： 由灰葡萄孢菌引起的番茄灰霉病是番茄主要病害之一，对番茄的产量和品质造成严重影响。为了对番茄灰霉病进行早期快速检测，以灰葡萄孢菌的ACTIN基因为靶基因，基于环介导等温扩增技术（Loop‐mediated isothermal amplification，LAMP），设计并筛选出一组LAMP特异性引物，优化其反应体系和反应条件，实现对灰葡萄孢菌的快速等温扩增。通过琼脂糖凝胶电泳和SYBR Green Ⅰ可视化分析，确定Bst Ⅱ DNA 聚合酶、dNTPs 的最佳用量和内外引物最佳比例分别为0.6 U/μL、1.25 mmol/L、2∶1，在61 ℃反应40 min即可特异性检测出灰葡萄孢菌，其灵敏度可达100 ag/μL，是普通PCR检测灵敏度的106倍。将该方法应用于番茄病害检测时，其对灰葡萄孢菌的孢子检出限达到20个/mL，且可在番茄被侵染4 d、尚无明显灰霉病感病表型的番茄叶片中检测出来，可用于番茄灰霉病的早期快速、灵敏、可视化检测。

关键词: 番茄, 灰葡萄孢菌, 灰霉病, 环介导等温扩增, ACTIN基因