甘蓝型油菜胞质体大量制备技术的研究

用Ｐｅｒｃｏｌｌ密度梯度离心法和吖啶橙（ＡＯ）结合光照处理两种方法，均从甘蓝型油菜下胚轴原生质体制备到胞质体。在２个Ｐｅｒｃｏｌｌ梯度、３００００ｇ（１８０００ｒ／ｍｉｎ）与１２℃离心６０ｍｉｎ的条件下，获得了含胞质体８０％以上的群体；以含８０、１００、１２０ｍｇ／Ｌ ＡＯ的酶液酶解下胚轴原生质体，纯化后分别给以３ｈ、２ｈ、１ｈ光照（光强度：４０００ｌｘ）可使原生质体几乎不能分裂，但保持５ｄ以上的

STUDIES ON BULK PRODUCTION OF CYTOPLASTS FROM HYPOCOTYL OF BRASSICA NAPUS

Bulk cytoplasts were produced from hypocotyl protoplasts of B. napus bymeans of a discontinuous Percoll density gradient centrifugation or treatment with acridine orange (AO)followed by exposure to light irradiation. More than 80% cytoplastswere obtained in the interphase between WS solution and 5% Percoll in a discontinuoustwo step gradients after 60 min centrafugation at 12 C 30 000 g(18 000 r/min). The protoplasts treated with 80 mg/L AO (14 -- 18 h during maceration) plus 3 h irradiation(4 000 lx), 100 mg/L AO plus 2 h irradiation or 120 mg/L AO plus 1 h irradiation seldom divided in culture,but could survive for more than 5 days. As light induced fragmentation of AO--DNA complex resulting in degradation of DNA,the nucleus inactivatedirreversibly,thus the products may by considered as cytoplasts.