Antibiotics for Agrobacterium Elimination in Apple Transformation

• Published in: Engeigaku kenkyuu Vol. 8; no. 4; pp. 419 - 426
• Main Authors: Komori, Sadao, Watanabe, Masano, Watanabe, Manabu, Tanaka, Norimitsu, Suzuki, Akira, Wada, Masato, Soejima, Junichi, Matsumoto, Shogo, Adachi, Yoshiteru, Li, Jijun
• Format: Journal Article
• Language: English; Japanese
• Published: Kyoto-City THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE 01.01.2009; Japan Science and Technology Agency
• Subjects: Agrobacterium tumefaciens; cefotaxime; Malus; Malus × domestica Borkh; meropenem; shoot regeneration
• ISSN: 1347-2658; 1880-3571
• DOI: 10.2503/hrj.8.419

We investigated several antibiotics including carbenicillin (CBPC), clavulanic acid/amoxicillin (CVA/AMPC), cefotaxime (CTX), meropenem (MEPM), vancomycin (VCM) and doxycycline (DOXY) for Agrobacterium elimination during apple transformation. The growth of Agrobacterium tumefaciens EHA101 was suppressed at 1,500 mg·L−1 of CBPC, 750 mg · L−1 of CVA/AMPC, less than 500 mg·L−1 of CTX, 50 mg·L−1 of MEPM and 200 mg · L−1 of DOXY, but growth was not suppressed at 1,500 mg·L−1 of VCM. In the shoot length, there was no difference among antibiotic-free, CBPC, CTX and VCM treatments. However, the shoot length in the high-concentration treatment of CVA/AMPC and DOXY was significantly shortened. However, the number of shoots increased as the concentration of MEPM rose. In CTX treatment, the shoot regeneration rate from leaf segments was lowered in comparison with antibiotic-free treatment; however, the regeneration rate was maintained to some degree even after high-concentration CTX treatment. The regeneration rate from leaf segments after MEPM treatment was superior to the antibiotic-free treatment. After DOXY treatment, leaf segments did not grow at all and finally died. These findings indicated that bacterial cell wall synthesis inhibitors such as CTX and MEPM were effective in eliminating Agrobacterium.