Interspecific hybridizations by electrofusion of embryogenic callus cells from `Seminole' tangelo (Citrus reticulata Blanco × C. paradisi Macf.), `Hazzara (Abohar)', or `Ohta' ponkan (C. reticulata Blanco) and leaf cells from `Lisbon' lemon [C. limon (L.) Burm. f.] or rough lemon (C. jambhili Lush.), respectively, were performed. Electrofusion of `Seminole' tangelo and `Lisbon' lemon, `Hazzara (Abohar)' and rough lemon, and `Ohta' ponkan and rough lemon resulted in 33, 43, and 36 plants, respectively. Seven to 10 plants in each combination were selected randomly and used to investigate nuclear and cytoplasmic genomes. Regenerated plants derived from electrofusion of `Seminole' tangelo and `Lisbon' lemon, and `Hazzara (Abohar)' and rough lemon possessed the same restriction fragment pattern for nuclear rDNA as that of the mesophyll parents and the same mitochondrial DNA (mtDNA) restriction pattern as that of the respective suspension parents, indicating that they were cybrids. In contrast, all the plants resulting from a combination between `Ohta' ponkan and rough lemon were confirmed by nuclear rDNA and mtDNA analysis to be somatic hybrids. The analysis of chromosome number supported the results of Southern blot hybridization. The results suggest that specific cell lines, parental combinations, or both can increase the efficiency of inducing cybrids in Citrus by electrofusion.
Somatic hybrids were produced by protoplast fusion between embryogenic callus cells of 'Hazzara (Abohar)' (Citrus reticulata Blanco) and mesophyll cells of Microcitrus australis (Planch.) Swingle, in an attempt to expand genetic diversity that might be valuable for Citrus breeding. Among the 40 plants acclimatized, 37 plants were intermediate in leaf morphology, whereas, the remaining 3 had leaves quite different from those of both parents. Ten plants among those with intermediate leaf morphology were selected for analysis of genome composition. These plants proved to be somatic hybrids by nuclear rDNA analysis. Mitochondrial DNA (mtDNA) analysis showed that all 10 hybrids contained 'Hazzara (Abohar)' plastome. Analysis of chloroplast DNA (cpDNA) showed that most of them carried specific fragments derived from 'Hazzara (Abohar)'. Others, however, carried specific fragments derived from both parents. Possible causes for such band patterns are discussed from a genetical viewpoint.
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