| 論文英文摘要 |
Gray mold caused by Botrytis cinerea has a severe impact on strawberry yield. This pathogen can infect various parts of the plant, including flowers, fruits, and leaves. Due to the broad host range and long latent period, infection often occurs during the flowering and immature fruit period, with symptoms appearing only after harvest. As a result, it is challenging to prevent and control the disease, leading to significant crop yield losses. Therefore, this study aimed to develop a rapid and labor-saving detection method for B. cinerea in strawberries using conventional polymerase chain reaction (PCR) and SYBR green-based real-time PCR techniques for Taiwanese farmers. This method can facilitate the rapid detection of asymptomatic infected fruits, providing an early diagnosis of the disease. According to literatures, the specific primers for B. cinerea, C729F/C729R, can be used in conjunction with conventional PCR to detect the pathogen, and another primer set, Rpb2aF/Rpb2aR, can detect gray mold pathogens on geraniums. However, the specificity of these two primer sets for B. cinerea strains in Taiwan has not been investigated, and a standardized molecular detection method for B. cinerea-infected strawberry fruit in Taiwan has not been established. Therefore, this study aims to verify the specificity of the primer sets C729F/C729R and Rpb2aF/Rpb2aR for B. cinerea strains in Taiwan, and then combine them with molecular detection techniques such as conventional PCR and SYBR green-based real-time PCR, as well as automated nucleic acid extraction methods, to construct a rapid and efficient detection process for B. cinerea-infected strawberry fruit. The results showed that the sensitivity of Rpb2aF/Rpb2aR in detecting samples (including standard DNA, genomic DNA, and conidia) was approximately ten times higher than that of C729F/C729R in both conventional PCR and SYBR green-based real-time PCR. Using the molecular detection methods established by combining the two primer sets with an automated nucleic acid extraction process, different levels of diseased strawberry fruits (including symptomless carrier fruits) could be detected in the SYBR green-based real-time PCR system. The detection rates of both primer sets for carrier samples were 100%, but the detection sensitivity of the method developed based on the Rpb2aF/Rpb2aR primer set was higher. Therefore, it is recommended to prioritize this primer set for the standardized molecular detection of Taiwan's strawberry gray mold carrier fruits using the SYBR green-based real-time PCR and automated nucleic acid extraction methods. This detection method can be applied to molecular detection after harvested strawberry. We hope to detect the pathogen before the occurrence of the disease in the future, formulate appropriate strategies for disease prevention or control, and reduce losses to strawberry fruit suppliers caused by the rapid spread of pathogens. |