ISSN 2736-1756
Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (7), pp. 001-006, July, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals
Full Length Research Paper
Rapid and Accurate Detection of Agrobacterium tumefaciens in Soil Using PCR
Wei Yang1,2, Lei Ji3, Li-Rong Tan2, Shi-Mo Li1, Yan Wang3, Hong-Xia Liu2*, and Yu-Ming Luo1*
1Jiangsu Key Laboratory of Eco-Agricultural Biotechnology around Hongze Lake, Huaiyin Teacher’s College, No. 111, Changjiang West, Huai’an, 223001, Jiangsu Province, China. 2Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. 3Shanghai Forestry Station, Shanghai, 200072, China.
Accepted 28 March, 2024
Abstract
One pair of primers was designed based on the sequence of tmr locus for specific and sensitive detection of Agrobacterium tumefaciens. Only the A. tumefaciens strain can produce the 236bp target fragment among the fourteen bacterial species that tested. The sensitivity of the specific PCR system was determined by a nested-PCR amplification which can numbered the copies of the template DNA. According to the results, it can give positive band when only 100 copies were in the template. The protocol was carried out for detection A. tumefaciens of twelve soil samples collected from six different gardens in Shanghai where crown gall happened. Two of the samples which collected from symptomless gardens also give the positive band. Based on the results we can make a conclusion that this pair of primers can be a useful tool in detecting A. tumefaciens, especially in detecting latent infection of this devastating pathogen.
Key words: Agrobacterium tumefaciens, detection, polymerase chain reaction (PCR).