蔬菜原生质体制备的实验方法研究

Study on experimental method of protoplast preparation of vegetables

原生质体融合是细胞工程课程实验不可或缺的内容,融合材料的选择以原生质体制备为基础。蔬菜种类丰富,价廉易得,使用安全,是原生质体制备和融合实验的良好材料。以八种蔬菜为原料,采用酶解法制备原生质体。经过初选和再选,确定以青椒和油菜作为融合观察实验的材料;并进一步对其原生质体制备过程中酶解液的种类和处理方法进行研究,确定青椒以13%CPW溶液为酶解液,油菜以2.5%KCl-1%MgSO_(4)溶液为酶解液,并均对酶解液进行热处理。

Protoplast fusion is an indispensable part of cell engineering course experiments.The selection of fusion materials is based on protoplast preparation.Vegetables are good materials for protoplast preparation and fusion experiments because of rich varieties,low price,getting easy and safety for use.Eight kinds of vegetables were used as raw materials to prepare protoplasts by enzymatic lysis.Green pepper and rape were determined as materials for research on fusion observation experiment by primary selection and re-selection.Furthermore,through the study of kinds of enzymatic hydrolysate and disposal method in protoplast preparation processes,it was determined that green pepper was treated with 13%CPW and rape was treated with 2.5%KCl-1%MgSO_(4) separately and both being heat-treated.