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| To evaluate the influences of the genotypes, anther developmental stages and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense. Methods The different effects such as four genotypes, plant growth regulators, and temperature condition were compared in the experiments. Results The most embryogenic calli was achieved for the genotypes 'ZCYH', 'Z4' and 'Z5' at the early- to middle-uninucleate stages, for genotype 'ZPM1' at the tetrad stage. Cold pretreatment increased the production of the embryogenic callus, and 4-d cold pretreatment improved the production of embryogenic callus from 0 to 2.2% and 5.0% for genotypes 'ZPM1' and 'ZCYH', respectively. No embryogenic callus were induced in the medium containing less than 0.75 mg/L 2,4-D. The highest regeneration rate (34.6%) was obtained in the half-strength MS salts regeneration medium supplemented with BA 0.1 mg/L. The low concentration BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis. Chromosome counting of regenerated plantlets showed mostly diploid plant (2n=12) with only one haploid plant (n=6). Because of the low rate of microspore embryo formation, we only track the process of embryogenesis from the connective tissue, instead of microspore by histological observations. Conclusion This study developed an efficient system embryogenic for callus induction and plant regeneration system from anthers of B. chinense. This is the first report on the haploid plantlet through the anther culture of B. chinense. |
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| Keywords:Medicobotany;Bupleurum chinense;anther culture;haploid |
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