Detection of the Causal Agent of Bacterial Wilt, Ralstonia solanacearum in the Seeds of Solanaceae by PCR

Cho, Jung Hee; Yim, Kyu Ock; Lee, Hyok In; Baeg, Ji Hyun; Cha, Jae Soon

Research in Plant Disease 2011;17(2):184-190.
Published online August 30, 2011.

가지과 종자에서 Ralstonia solanacearum의 검출을 위한 PCR 방법

조정희, 임규옥, 이혁인, 백지현, 차재순

Detection of the Causal Agent of Bacterial Wilt, Ralstonia solanacearum in the Seeds of Solanaceae by PCR

Jung Hee Cho, Kyu Ock Yim, Hyok In Lee, Ji Hyun Baeg, Jae Soon Cha

Abstract

Ralstonia solanacearum, a causal agent of bacterium wilt is very difficult to control once the disease becomes endemic. Thus, Ralstonia solanacearum is a plant quarantine bacterium in many countries including Korea. In this study, we developed PCR assays, which can detect Ralstonia solanacearum from the Solanaceae seeds. Primers RS-JH-F and RS-JH-R amplified specifically a 401 bp fragment only from Ralstonia solanacearum race 1 and race 3. The nested PCR primers, RS-JH-F-ne and RS-JH-R-ne that were designed inside of 1st PCR amplicon amplified specifically a 131 bp fragment only from Ralstonia solanacearum race 1 and race 3. The primers did not amplify any non-specific DNA from the seed extracts of the Solanaceae including tomato and pepper. When detection sensitivity were compared using the Solanaceae seeds inoculated with target bacteria artificially, the nested PCR method developed in this study 100 times more sensitive than ELISA and selective medium. Therefore, we believe that the PCR assays developed in this work is very useful to detect Ralstonia solanacearum in the Solanaceae seeds.

Key Words: PCR assays, Plant quarantine, Seed detection