Thidiazuron required for efficient somatic embryogenesis from suspension-cultured cells ofPimpinella brachycarpa |
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Authors: | Joon Chul Kim Mi Young Chang Su In Son Su Jeong Heo |
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Institution: | (1) Division of Biological Science, College of Natural Sciences, Kangwon National University, 200-701 Chunchon, Korea;(2) Regional Crop Experiment Station, Kangwon Agricultural Research and Extension Services, c00-820 Chunchon, Korea |
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Abstract: | To maintain embryogenic cell lines ofPimpinella brachycarpa, we suspension-cultured friable and rapidly growing yellowish calli in an MS liquid medium containing 0.2 ~ 2,4-D and 0.5pM BAP. Efficient somatic embryogenesis was achieved when selected cells were then transferred to an MS medium (0.2% gelrite)
that contained 0.2gM 2,4-D, 0.5 uM BAP, and 10.0 laM TDZ (thidiazuron). These cells were cultured at 27°C under continuous illumination (21.5
I~E m-2 s-l). Embryogenic calli expanded about four-fold, and developed into pale yellow calli. Somatic embryogenesis was initiated only
from glossy and nodular-type calli. After two more weeks of culture, globular embryos appeared on the surface of calli grown
in the MS medium that contained 10.0 /aM TDZ only, or in combination with 0.5 gM NAA. Experimenting with 2,4-D, an auxin,
to promote embryogenic calli resulted in excessive browning and death. We overcame this problem by growing glossy embryogenic
and nodular calli on media that contained 10.0 gM TDZ. Calli that were not treated with TDZ turned dark brown and were not
viable. Up to 74% of the calli showed somatic embryos when the medium was supplemented with 10.0 uM TDZ and 0.5 uM NAA. Embryos
from these TDZ-induced, somatic embryogenic calli grew efficiently, forming multiple shoots and developing into normal plants.
Therefore, efficient differentiation of suspension-cultured cell clusters into embryogenic calli, along with treatment of
subsequent somatic embryos by TDZ, suggests that TDZ probably helps in establishing the optimum cytokinin-auxin ratio required
for induction and expression of somatic embryogenesis. |
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Keywords: | differentiation embryogenic calli globular embryo multiple shoots plantlet thidiazuron |
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