Abstract
We have developed a new Agrobacterium-mediated transformation method for the low-frequency-regenerating pear (Pyrus communis L.) cvs. Silver bell and La France. Leaf sections derived from in vitro shoots were initially used for the transformation procedure. Under optimum transformation conditions, which included culture and selection on 30 mg/l kanamycin (Km) combined with 500 mg/l sulbenicillin, a 3.2% transformation efficiency was obtained for cv. Silver bell, but no transformants of La France were obtained because of the very low regeneration frequency. Axillary shoot meristems were then examined as potential explants for La France. Selection in 5 mg/l Km and 375 mg/l carbenicillin resulted in transformed shoots being produced at an efficiency of 4.8%, and the apparent white Km-sensitive shoots were not formed during a 2-year subculture on micropropagation medium containing 50 mg/l Km. Therefore, transformations using axillary shoot meristems may be an alternative method for pear cultivars recalcitrant to regeneration from leaf sections.
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Abbreviations
- BA:
-
6-Benzylaminopurine
- CaMV 35S:
-
35S RNA of cauliflower mosaic virus
- Cb:
-
Carbenicillin
- Cf:
-
Cefotaxime
- GUS:
-
β-Glucuronidase
- hpt :
-
Hygromycin phosphotransferase gene
- Hyg:
-
Hygromycin B
- Km:
-
Kanamycin
- NAA:
-
α-Naphthaleneacetic acid
- nptII:
-
Neomycin phosphotransferase II gene
- Sb:
-
Sulbenicillin
- TDZ:
-
Thidiazuron
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Acknowledgements
This study was supported, in part, by the Program for the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) and the Regional Joint Research Project of Yamagata Prefecture. The practical work was conducted at the Yamagata Prefectural Horticultural Experiment Station. We thank all of the members of the station for their kind assistance. We also sincerely thank our Assistant Researchers Atsuko Saito, Kayoko Komuro, and Keiko Watanabe for their assistance with the culture work.
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Matsuda, N., Gao, M., Isuzugawa, K. et al. Development of an Agrobacterium-mediated transformation method for pear (Pyrus communis L.) with leaf-section and axillary shoot-meristem explants. Plant Cell Rep 24, 45–51 (2005). https://doi.org/10.1007/s00299-005-0924-1
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DOI: https://doi.org/10.1007/s00299-005-0924-1