Development of an Agrobacterium-mediated transformation method for pear (Pyrus communis L.) with leaf-section and axillary shoot-meristem explants

• Published in: Plant cell reports Vol. 24; no. 1; pp. 45 - 51
• Main Authors: Matsuda, N, Gao, M, Isuzugawa, K, Takashina, T, Nishimura, K
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.04.2005
• Subjects: Agrobacterium; Agrobacterium tumefaciens; Biological and medical sciences; Biotechnology; Eukaryotic cell cultures; explants; fruit trees; Fundamental and applied biological sciences. Psychology; Genetic Engineering - methods; genetic transformation; genetic vectors; Kanamycin; leaves; Methods. Procedures. Technologies; micropropagation; Miscellaneous; molecular sequence data; nucleotide sequences; pears; Plant cells and fungal cells; Plant Leaves - genetics; Plant Leaves - growth & development; Plant Shoots - genetics; Plant Shoots - growth & development; Plants, Genetically Modified; Pyrus - genetics; Pyrus - growth & development; Pyrus communis; shoot meristems; Transformation, Genetic; Pyrus communis; Vegetals; Transformation; Rosaceae; Leaf section; Pyri communis L; Plant leaf; Meristem; Axillary shoot meristem; Regeneration; Agrobacterium; Dicotyledones; Angiospermae; Bacteria; Spermatophyta; Rhizobiaceae; Shoot
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-005-0924-1

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.