caa a22 4500 469027193 CHVBK 20180323132733.0 cr unu---uuuuu 170328e19921201xx s 000 0 eng 10.1007/BF00232413 doi (NATIONALLICENCE)springer-10.1007/BF00232413 Efficient transformation of Arabidopsis thaliana [Elektronische Daten] comparison of the efficiencies with various organs, plant ecotypes and Agrobacterium strains [Kazuhito Akama, Hideaki Shiraishi, Shozo Ohta, Kenzo Nakamura, Kiyotaka Okada, Yoshiro Shimura] Summary: The efficiency of Agrobacterium-mediated transformation of Arabidopsis thaliana was compared with different organs, Arabidopsis ecotypes, and Agrobacterium strains. Efficiency of shoot regeneration was examined using hypocotyl, cotyledon and root explants prepared from young seedlings. Hypocotyl expiants had the highest regeneration efficiency in all of the four Arabidopsis ecotypes tested, when based on a tissue culture system of callus-inducing medium (CIM: Valvekens et al. 1988) and shoot-inducing medium (SIM: Feldmann and Marks 1986). Histochemical analysis using the ß-glucuronidase (GUS) reporter gene showed that the gusA gene expression increased as the period of preincubation on CIM was extended, suggesting that dividing cells are susceptible to Agrobacterium infection. In order to obtain transgenic shoots, hypocotyl explants preincubated for 7 or 8 days on CIM were infected with Agrobacterium containing a binary vector which carries two drug-resistant genes as selection markers, and transferred to SIM for selection of transformed shoots. Of four Arabidopsis ecotypes and of three Agrobacterium strains examined, Wassilewskija ecotype and EHA101 strain showed the highest efficiency of regeneration of transformed shoots. By combining the most efficient factors of preincubation period, Arabidopsis ecotype, tissue, and bacterial strain, we obtained a transformation efficiency of about 80-90%. Southern analysis of 124 transgenic plants showed that 44% had one copy of inserted T-DNA while the others had more than one copy. Springer-Verlag, 1992 Arabidopsis thaliana nationallicence Transformation nationallicence Agrobacterium nationallicence T-DNA nationallicence Shoot regeneration nationallicence Transgenic plant nationallicence AIM : Agrobacterium infection medium nationallicence CIM : callus-inducing medium nationallicence CTAB : cetyltrimethylammonium bromide nationallicence 2,4-D : 2,4-dichlorophenoxy-acetic acid nationallicence GUS : ß-glucuronidase nationallicence hph : hygromycin phosphotransferase nationallicence IAA : indole-3-acetic acid nationallicence IBA : indole-3-butyric acid nationallicence 2ip : N -(2-isopentenyl) adenine nationallicence NPTII : neomycin phosphotransferase II nationallicence RIM : root-inducing medium nationallicence 35S : cauliflower mosaic virus 35S promoter nationallicence SIM : shoot-inducing medium nationallicence Akama Kazuhito Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut Shiraishi Hideaki Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut Ohta Shozo Laboratory of Biochemistry, School of Agriculture, Nagoya University, 464-01, Nagoya, Japan aut Nakamura Kenzo Laboratory of Biochemistry, School of Agriculture, Nagoya University, 464-01, Nagoya, Japan aut Okada Kiyotaka Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut Shimura Yoshiro Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut Plant Cell Reports Springer-Verlag 12/1(1992-12-01), 7-11 0721-7714 12:1<7 1992 12 299 https://doi.org/10.1007/BF00232413 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00232413 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Akama Kazuhito Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut NATIONALLICENCE 700 1- Shiraishi Hideaki Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut NATIONALLICENCE 700 1- Ohta Shozo Laboratory of Biochemistry, School of Agriculture, Nagoya University, 464-01, Nagoya, Japan aut NATIONALLICENCE 700 1- Nakamura Kenzo Laboratory of Biochemistry, School of Agriculture, Nagoya University, 464-01, Nagoya, Japan aut NATIONALLICENCE 700 1- Okada Kiyotaka Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut NATIONALLICENCE 700 1- Shimura Yoshiro Division 1 of Gene Expression and Regulation, National Institute for Basic Biology, 444, Okazaki, Japan aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 12/1(1992-12-01), 7-11 0721-7714 12:1<7 1992 12 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer