关键词: 番茄, 灰葡萄孢菌, 灰霉病, 环介导等温扩增, ACTIN基因

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