Protoplasts of Valencia sweet orange ( Citrus sinensis Osb.),irradiated by X_ray with a dose rate of 3.8 krad/min for 45 min, were electrically fused with protoplasts of Murcott tangor ( C. reticulata×C. sin...Protoplasts of Valencia sweet orange ( Citrus sinensis Osb.),irradiated by X_ray with a dose rate of 3.8 krad/min for 45 min, were electrically fused with protoplasts of Murcott tangor ( C. reticulata×C. sinensis ) that were treated with 0.25 mmol/L iodoacetic acid for 15 min. It took nearly 15 months for the fusion_derived calli to develop into embryoids that were only originated in the medium of MT supplemented with 2% glycerol. The shoots were recalcitrant to rooting in the root_induction medium. In vitro grafting was employed to produce whole plants though one self_rooting plant was obtained. Cytological determination of root and shoot tips showed mainly diploid and aneuploid cells, together with few tetraploid cells in some plants. RAPD (random amplified polymorphism DNA) analysis with 10_mer primers demonstrated that bands specific to the fusion parents were detected in the regenerated plants, indicating that interspecific somatic hybrids have been obtained via protoplast asymmetric fusion in Citrus .展开更多
RFLP (restriction fragment length polymorphism) was employed to analyze cytoplasmic genome of diploid somatic hybrid plant, morphologically similar to rough lemon which was leaf parent, that was produced via protoplas...RFLP (restriction fragment length polymorphism) was employed to analyze cytoplasmic genome of diploid somatic hybrid plant, morphologically similar to rough lemon which was leaf parent, that was produced via protoplast fusion between rough lemon ( Citrus jambhiri Lush) and Hamlin sweet orange ( C. sinensis Osb.), the embryogenic parent. Three enzyme_mitocondrial probe combinations and one enzyme_chloroplast probe combination demonstrated that the plant had identical band profiles to Hamlin sweet orange as far as mtDNA and cpDNA were concerned.展开更多
Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of ...Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band patterns to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, Whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.展开更多
基金TheNational Natural Science Foundation of China(3983026039625017)the International Foundation for Science(D/3001_1)
文摘Protoplasts of Valencia sweet orange ( Citrus sinensis Osb.),irradiated by X_ray with a dose rate of 3.8 krad/min for 45 min, were electrically fused with protoplasts of Murcott tangor ( C. reticulata×C. sinensis ) that were treated with 0.25 mmol/L iodoacetic acid for 15 min. It took nearly 15 months for the fusion_derived calli to develop into embryoids that were only originated in the medium of MT supplemented with 2% glycerol. The shoots were recalcitrant to rooting in the root_induction medium. In vitro grafting was employed to produce whole plants though one self_rooting plant was obtained. Cytological determination of root and shoot tips showed mainly diploid and aneuploid cells, together with few tetraploid cells in some plants. RAPD (random amplified polymorphism DNA) analysis with 10_mer primers demonstrated that bands specific to the fusion parents were detected in the regenerated plants, indicating that interspecific somatic hybrids have been obtained via protoplast asymmetric fusion in Citrus .
文摘RFLP (restriction fragment length polymorphism) was employed to analyze cytoplasmic genome of diploid somatic hybrid plant, morphologically similar to rough lemon which was leaf parent, that was produced via protoplast fusion between rough lemon ( Citrus jambhiri Lush) and Hamlin sweet orange ( C. sinensis Osb.), the embryogenic parent. Three enzyme_mitocondrial probe combinations and one enzyme_chloroplast probe combination demonstrated that the plant had identical band profiles to Hamlin sweet orange as far as mtDNA and cpDNA were concerned.
文摘Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band patterns to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, Whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.
