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KMID : 1098420070150010046
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Korean Journal of Medicinal Crop Science
2007 Volume.15 No. 1 p.46 ~ p.50
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Optimal Culture Conditions for Transformed Root Growth and Trichosanthin Formation in Trichosanthes kirilowii Max.
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Hwang Sung-Jin
Na Myung-Suk
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Abstract
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Transformed hairy roots were induced from in vitro grown plantlets of Trichosanthes kirilowii by infection with Agrobacterium rhizogenes strain ATCC15834. Transformed hairy roots exhibited active growth with high branching of roots on plant growth regulators-free medium. Cloned line (TR-03) of hairy root was tested for its growth and extracellular protein accumulation in medium under various culture conditions. Among the culture media tested, a full-strength MS medium had a pronounced effect on root biomass and extracelluar protein accumulation in medium. The maximum root biomass (2.4 g DRW/flask) and extracellular total protein contents (28.3ug/mell) in medium was obtained at inoculum size of 2 g (FRW) and in MS medium supplemented with 4% sucrose. In addition, the optimal shaking speed for root growth and extracellular protein accumulation in medium were 100 rpm. The total extracellualr protein concentration reached a maximum of 28.3ug/mell at 4 weeks and decreased thereafter. Protein translation inhibitory activity was observed in culture broths and reached levels of 21.3 unit. These studies demonstrate that the transformed hairy roots can be utilized for the in vitro production of ribosome-inactivating proteins.
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KEYWORD
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trichosanthin, biomass, hairy root clone, Trichosanthes kirilowii
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