Somatic hybrids obtained by fusion betweenPoncirus trifoliata (2x) andFortunella hindsii (4x) protoplasts |
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| Authors: | M. Miranda T. Motomura F. Ikeda T. Ohgawara W. Saito T. Endo M. Omura T. Moriguchi |
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| Affiliation: | (1) Faculty of Agriculture, Tokyo University of Agriculture, 156 Setagaya-ku, Tokyo, Japan;(2) Shoda Shoyu Co., 374 Tatebayashi, Gunma, Japan;(3) Research and Developmental Division, Kikkoman Corporation, 278 Noda, Chiba, Japan;(4) Department of Citriculture, National Institute of Fruit Tree Science, Okitsu, 424-02 Shimizu, Shizuoka, Japan |
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| Abstract: | Somatic hybrids were obtained by the symmetric fusion of embryogenic callus cells from tetraploid  Mame kumquat [Fortunella hindsii (Champ.) Swing.] and mesophyll cells from diploid trifoliate orange [Poncirus trifoliata (L.) Raf.]. Southern blot analysis of three regenerants revealed that they carried specific rDNA fragments from both fusion partners, thereby confirming their hybridity. In contrast, mitochondrial DNA (mtDNA) and chloroblast DNA (cpDNA) were unidirectionally transmitted from the callus parent without any evidence of recombination. No differences in the restriction fragment patterns of rDNA, mtDNA or cpDNA could be detected among the regenerants. Flow cytometry showed that two regenerants were hexaploids, as expected, but that one was pentaploid, probably due to elimination of chromosomes prior to the regeneration of this plant.Abbreviations MS Murashige and Skoog (1962) - DIG digoxigenin - AMPPD 3-(2 -spiroadamantane)-4-methoxy-4-(3 -phosphoryloxy)-phenyl-1,2-dioxetane - FACS fluorescence-activated cell sorter |
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| Keywords: | Electrofusion Flow cytometry Chromosome elimination Polyploidy Citrus |
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