Roots of Codonopsis pilosula (Franch.) Nannf. are among the most popular Chinese herbal medicines, exhibiting various beneficial activities which support immunity and stress resistance. The plant shows high intraspecific genetic variation. There is a need for effective vegetative propagation methods yielding high and sustainable quality. Here we report a micropropagation method using axillary shoot proliferation. Nodal segments from aseptically germinated plants were inoculated on modified MS media enriched with different concentrations of cytokinins: benzyladenine, kinetin (1, 4, 10 or 20 μM) or thidiazuron (1, 4 or 8 μM), with or without the auxin NAA (1 μM). Axillary bud break was initiated most efficiently on media with 1 or 4 μM BA and 1μM NAA. Shoot number increased markedly in subsequent cycles of harvesting and transfer to fresh 1 μM BA and NAA medium, leading to the maximum 69 shoots (mean 38.16±4.35) from a single nodal explant in the fourth harvest. The shoots were successfully (>98% efficiency) rooted in MS medium containing high sucrose (60 g/L) and 5 μM IAA, and acclimatized to soil cultivation with a survival rate of 90%. These results can be used to establish a simple and commercially viable protocol for mass propagation of C. pilosula for plantations or breeding.
This communication reports detection of somaclonal variation among tissue culture-raised plants of Amorphophallus rivieri Durieu, an economically important crop in China, with high content of glucomannan in its corms. A population of regenerated plants was obtained from a single donor plant of A. rivieri via corm organogenesis, and 28 plants were randomly selected as a representative sample and subjected to analysis of somaclonal variation using inter-simple sequence repeat (ISSR) markers. Of the 26 ISSR primers screened, 13 gave distinct and reproducible band patterns, yielding 131 bands with an average of 10.1 bands per primer. Ten primers were polymorphic and generated 16 polymorphic bands with 12.2% mean polymorphism. Based on the ISSR data from the regenerated plants and the donor plant, Jaccard's similarity coefficients were calculated; they ranged from 0.961 to 1.000 with a mean of 0.982. A dendrogram was constructed using the unweighted pair group method with arithmetic mean (Upgma); it showed that a majority of regenerated plants (including the donor plant) clustered closely, with a mean similarity coefficient of 0.987. Low somaclonal variation observed in the regenerated plants indicates that rapid propagation of A. rivieri via corm organogenesis is a practicable method with a low risk of genetic instability.