caa a22 4500 463160606 CHVBK 20180406164746.0 cr unu---uuuuu 170326e20070601xx s 000 0 eng 10.1007/s10535-007-0043-7 doi (NATIONALLICENCE)springer-10.1007/s10535-007-0043-7 High efficient transgenic plant regeneration from embryogenic calluses of Citrus sinensis [Elektronische Daten] [Y. Duan, W. Guo, H. Meng, N. Tao, D. Li, X. Deng] Transformation and high efficient regeneration of transgenic plants from embryogenic calluses of Bingtang sweet orange [Citrus sinensis (L.) Osbeck] was reported. Embryogenic calluses were inoculated with Agrobacterium tumefaciens strain EHA105, harboring the binary Ti plasmid pROK II and carrying a neomycin phosphotransferase II (NPTII) gene, an intron β-glucuronidase (GUS) gene and the Arabidopsis APETALA1 (AP1) gene. Transformation treatment was with inoculation time of 30 min, co-culture of 3 d at 23 °C and supplementation of the co-culture medium with 2 mg dm−3 acetosyringone (AS). Kanamycin (50 mg dm−3) was effective to inhibit the growth of non-transformed calluses while it did not affect the transformed ones. The total number of transformed callus lines was 7 with 100 % embryo induction. High efficient regeneration of the transgenic embryos (88 % with 4-5 shoots per embryoid) was realized within 3 months. Integration of the transgene into the citrus genome was confirmed by histochemical GUS staining, polymerase chain reaction (PCR) analysis with AP1-specific primer and Southern blot hybridization with a 712 bp PCR fragment of AP1 as the probe. Institute of Experimental Botany, ASCR, 2007 Agrobacterium tumefaciens nationallicence Bingtang sweet orange nationallicence GUS nationallicence kanamycin nationallicence PCR nationallicence plantlet regeneration nationallicence Southern blot nationallicence transformation nationallicence AS : acetosyringone nationallicence BA : 6-benzylaminopurine nationallicence Cef : cefotaxime nationallicence Cm : chloromycetin nationallicence GUS : β-glucuronidase nationallicence Kin : kinetin nationallicence Km : kanamycin nationallicence LB medium : Luria-Bertani medium nationallicence MT : Murashige and Tucker nationallicence NAA : α-naphthaleneacetic acid nationallicence NPTII : neomycin phosphotransferase II nationallicence PCR : polymerase chain reaction nationallicence Duan Y. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Guo W. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Meng H. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Tao N. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Li D. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Deng X. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/2(2007-06-01), 212-216 0006-3134 51:2<212 2007 51 10535 https://doi.org/10.1007/s10535-007-0043-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10535-007-0043-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Duan Y. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 700 1- Guo W. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 700 1- Meng H. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 700 1- Tao N. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 700 1- Li D. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 700 1- Deng X. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China aut NATIONALLICENCE 773 0- Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/2(2007-06-01), 212-216 0006-3134 51:2<212 2007 51 10535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer