Somatic hybridization between Citrus reticulata and Citropsis gabunensis through electrofusion |
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Authors: | Jing-Tian Ling M. Iwamasa |
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Affiliation: | (1) Faculty of Agriculture, Saga University, 840 Saga, Japan;(2) Tennessee State University, 3500 John A. Merritt Blvd, P. O. Box 1322, 37209-1561 Nashville, TN, USA;(3) Present address: Faculty of Agriculture, Kyushu Toukai University, 849-14 Kumamoto, Japan |
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Abstract: | Protoplasts from embryogenic calli of Citrus reticulata Blanco cv. Ponkan and Citropsis gabunensis (Engl.) Swing. & M. Kell (Cabon Cherry Orange), were isolated and fused using electric current. Maximum fusion frequency was obtained with AC at 75 kV/cm (1.0 MHz) for 15 s, followed by DC square-wave pulses at 1.25 kV/cm for 40 s. Fusion-treated protoplasts were cultured on MT medium containing no growth regulators, solidified with 0.6% Bacto Difco agar. Protoplast-derived calli were proliferated on MT medium containing 1 mg/l zeatin and 0.9% agar. A total of 31 lines of somatic hybrid calli were obtained by screening on the basis of chromosome count and isozyme analysis. The somatic hybrids were tetraploid (2n=36). Plants were regenerated from the calli via somatic embryogenesis. The somatic hybrid plants exhibited morphological characteristics intermediate to the parental plants.Abbreviations 2,4-D 2, 4-dichlorophenoxyacetic acid - BA 6-benzylaminopurine - MT Murashige and Tucker (1969) - PEG polyethylene glycol - AC alternating current - DC direct current |
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