Cell Growth in Suspension-Culture of Populus nigra var. italica and the Efficiency of Micro-Callus Formation according to Cell Plating Method

Populus nigra var. italica현탁배양(懸濁培養) 세포(細胞)의 생장(生長) 및 Cell Plating방법(方法)에 따른 Micro-Callus형성능력(形成能力)

In order to know the growth of suspended cells by explant sources, the change of nitrogen contents of cultured cells following the growth periods, capability of micro-callus formation according to cell plating methods, growth of suspended cells on various media, and efficiency of micro-callus formation by using growth regulators and different N strengths were investigated. 1. When suspension culture was tried by using the callus induced from internode and petiole, cell fresh weight and packed cell volume increased with similar way and the growth reached at stationary phase after 12 culture days. 2. N-contents of cultured cells increased upto 3 days and decreased around 6days. But the values increased again upto 9 days, after that they showed gradual decreases. 3. Of cell plating methods, embedding method was the best for micro-callus formation. 4. Growth of suspened cells showed the rest performanoes, when they were cultured on LM medium with 1/2N strengths and BAP 0.01.2.4-D 0.1, and NAA $1.0mg/{\ell}$, after 15 cultured days(upto 76.9 folds). LM medium was better than MS or GD. The combination of auxin and cytokinin was better for cell growing than auxin-treatment only. 5. Micro-callus from single cell and small cell aggregates was formed only on MS and LM media with 2,4-D $1.0mg/{\ell}$.

양버들의 explant soure에 따른 현탁배양(懸濁培養) 세포(細胞)의 생장(生長)과 생장기간(生長其間)에 따른 배양세포내(培養細胞內) 질소함량(窒素含量) 변화(變化), plating방법(方法)에 따른 micro-callus 형성능력(形成能力), 배양배지(培養培地)의 종류(種類) 및 생장조절물질(生長調節物質), 질소량(窒素量)에 따른 현탁배양세포(懸濁培養細胞)의 생장(生長)과 고체배지(古體培地)에 plating하였을 때 micro-callus 형성(形成)을 조사(調査)하여 다음과 같은 결과(結果)를 얻었다. 1. 절간조직(節間組織)과 엽병조직(葉柄組織)에서 유래(由來)된 callus를 재료(材料)로 하여, 현탁배양(懸濁培養)하였을 때 세포(細胞)의 생중량(生重量)과 packed cell volume은 거의 비슷하게 증가(增加)하였으며, 배양후(培養後) 12일(日)경에 stationary phase에 도달하였다. 2. 배양세포(培養細胞)의 N함량(含量)은 배양후(培養後) 3일(日)경에 높아졌으며, 6일(日)경에는 낮아지고, 다시 9일(日)까지 높아지다가 그 이후(以後)는 서서히 떨어지는 경향을 나타내었다. 3. Cell plating에서 embedding방법(方法)이 가장 높은 micro-callus형성율(形成率)을 보였다. 4. 현탁배양세포(懸濁培養細胞)의 생장(生長)은 LM배지(培地)의 질소량(窒素量)을 1/2로 하고 BAP 0.01, 2,4-D0.1, NAA $1.0mg/{\ell}$ 첨가한 배지(培地)에서 배양(培養) 15일 후에 76.9배(倍)의 생장량(生長量)을 보여 가장 양호(良好)하였고, LM 배지(培地)가 MS나 GD배지(培地)보다 생장(生長)이 좋았으며, auxin만 사용(使用)한 것보다 cytokinin과 혼합하여 배양(培養)하였을 때, 더 좋은 세포생장(細胞生長)을 나타내었다. 5. Single cell 및 small cell aggregates 집단(集團)으로부터의 micro-callus 형성(形成)은 2,4-D를 $1.0mg/{\ell}$ 첨가한 LM 및 MS배지(培地)에서 배양(培養)하였을 때만 형성(形成)되었다.