Studies on the Callus Culture of Stevia as a New Sweetening Source and the Formation of Stevioside

신감미자원식물(新甘味資源植物) 스테비아의 Callus 배양(培養)과 Stevioside 생성(生成)에 관(關)한 연구(硏究)

This experiment was carried out to clarify the optimal concentrations of growth regulators for callus induction and the condition of callus culture of leaf tissue taken from Stevia rebaudiana Bertoni. The content of stevioside, sweetening component, in leaf-derived callus of stevia was also investigated. It was shown that the optimal concentrations of benzyladenine (BA) and ${\alpha}-naphthalene$acetic acid (NAA) for callus induction were $10^{-6}M$ and $10^{-5}M$, respectively. Reculture of these calli in media (Linsmaier and Skoog) supplemented with BA $10^{-6}M$ and NAA $10^{-5}M$ resulted in profuse calli 15 to 20 days after incubation. When sweetening components produced by callus were extracted and identified by TLC, stevioside appeared to have Rf value 0.50 in TLC which was exactly same as standard stevioside. Stevioside content obtained by TLC-FID analyzer was 260mg per 100g on the basis of dry weight.

Stevia엽(葉) 조직(組織)에서 callus의 유기(誘起) 및 배양(培養)을 위(爲)한 조건(條件)을 조사(調査)하고 이들 배양(培養) callus가 감미성분(甘味成分)인 stevioside를 생성(生成)하는지를 조사(調査)하여 다음과 같은 결과(結果)를 얻었다. Callus유기(誘起)에는 Linsmaier-Skoog기본배지(基本培地)에 benzyl-adenine $10^{-6}M$과 ${\alpha}-naphthalene$ acetic acid $10^{-5}M$을 함유(含有)한 배지(培地)가 가장 효과적(效果的)이였으며 callus의 성장률(成長率)도 비교적 양호(良好)하였으며 이들 callus는 배양후(培養後) 15일(日)부터 20일(日) 사이에 가장 빠른 성장률(成長率)을 나타내었다. Stevia callus가 생성(生成)한 stevioside는 TLC에 의(依)해 동정(同定)하였으며, 이는 정제(精製)한 표준품(標準品)의 stevioside와 동일(同一)한 Rf값인 0.50을 나타내었다. 또한 stevioside을TLC-FID analyzer에 의(依)해 분석(分析)한 결과(結果), 배양(培養) callus 건물중(乾物重) 100g당(當) 260mg이였다.