ABSTRACT
The embryo-derived calli from four types of tall fescues (Festuca arundinacea Schreb) were transformed with two Agrobactrium tumefaciens strains AGL1 and GV3101. AGL1 harbors an intron-AtNHX1 expression vector pROK2U containing ubiqutin promoter and npt II marker gene. GV3101 harbors an intron-AtNHX1 expression vector pROK2 containing 35S promoter and npt II gene. After infection and co-culture with AGL1 or GV3101, the embyogenic calli were selected with 50-150 mg/L paromomycine and 1126 resistant plants regenerated from the resistant calli. All plants were selected further with 10-20 mg/L Kanamycin and 525 of them remained green. Genome DNA of the resistant plants was checked with specific primers and probe from AtNHX1 gene. The results of PCR assay and Southern blot analysis indicted that exogenous target gene (AtNHX1 gene) had been transferred into different cultivars of Festuca arundinacea. Different transformation frequencies among the four cultivars were obtained.
Subject(s)
Agrobacterium tumefaciens/genetics , Festuca/genetics , Transformation, Genetic , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cation Transport Proteins/genetics , Cation Transport Proteins/metabolism , DNA, Plant/genetics , Festuca/physiology , Introns , Plants, Genetically Modified , Promoter Regions, Genetic , Regeneration , Sodium-Hydrogen Exchangers/genetics , Sodium-Hydrogen Exchangers/metabolism , Sodium-Hydrogen Exchangers/physiology , Tissue Culture Techniques , Ubiquitin/geneticsABSTRACT
We report the production and characterization of somatic hybrids between Triticum aestivum L. and Agropyron elongatum (Host) Nevishi (the synonym is Thinopyrum ponticum). Asymmetric protoplast fusion was performed between Agropyron elongatum protoplasts irradiated with a low UV dose and protoplasts of wheat taken from nonregenerable suspension cultures. More than 40 green plantlets were obtained from 15 regenerated clones and one of them produced seeds. The phenotypes of the hybrid plants and seeds were intermediate between wheat and Agropyron elongatum. All of the regenerated calli and plants were verified as intergeneric hybrids on the basis of morphological observation and analysis of isozyme, cytological, 5SrDNA spacer sequences and random amplified polymorphic DNA (RAPD). RFLP analysis of the mitochondrial genome revealed evidence of random segregation and recombination of mtDNA.
Subject(s)
Agropyron/genetics , Hybridization, Genetic/radiation effects , Triticum/genetics , Ultraviolet Rays , Agropyron/radiation effects , Blotting, Southern , Cell Fusion , Chromosomes, Plant/chemistry , Chromosomes, Plant/genetics , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/genetics , DNA, Plant/chemistry , DNA, Plant/genetics , DNA, Ribosomal Spacer/analysis , Dose-Response Relationship, Radiation , Electrophoresis, Polyacrylamide Gel , Esterases/analysis , Fertility/genetics , Fertility/radiation effects , Genotype , Hybrid Cells/chemistry , Hybrid Cells/cytology , Hybrid Cells/enzymology , Isoenzymes/analysis , Peroxidase/analysis , Phenotype , Plant Development , Plants/anatomy & histology , Plants/chemistry , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Protoplasts/cytology , Protoplasts/radiation effects , Random Amplified Polymorphic DNA Technique , Recombination, Genetic/genetics , Seeds/anatomy & histology , Triticum/radiation effectsABSTRACT
OBJECTIVE: To transfer the effective elements of Bupleurum scorzonerifolium into carrot, and provide theoretical data for the exploitation, improvement and selection of the germplasm of Chinese medicinal plants. METHOD: The protoplasta of Bupleurum scorzonerifolium irradiated by ultraviolet light (UV) at an intensity of 300 microW.(cm2)-1 for 0, 1, 2 min respectively were fused with those of carrot Fisch by PEG method. The regenerated clones, derived form a single fused cell, were examined for their hybrid nature by phenotype and Esterase isoenzyme analysis. RESULT: Nine clones were identified as the somatic hybrids between B. scorzonerifolium and carrot. CONCLUSION: This provides a firm foundation for the further analysis of the main active components saikosaponin of somatic hybrids and the screening out of high-medicine-content hybrid cell lines.
