摘要
本文以钙黄素为荧光标记物,使用电击法,研究了不同条件,如浸种液、苗龄、培养基、酶解时间等和电击参数对大麦叶肉原生质体的分离得率及电击导入外源荧光物质的影响。通过电击成功地将荧光标记物钙黄素导入大麦叶肉原生质体内。上述条件和电击参数对获得有活力的大麦叶肉原生质体的数量及电击导入钙黄素的频率均有明显的影响。这些实验结果为进一步进行电击导入GUS等外源基因于大麦叶肉原生质体内、并实现瞬间表达的研究打下了实验基础。
In this report,we have studied the effects of the status of barley mesophyll protoplasts on the entrance of calcein by electroporation. The results are follows.DSeed-saoking with S-3307 and adding the nutritional solution of 1/4 MS in medium during grow seedlings were remarkably favourable to breed healthy and strong seedlings of barley andlthereby to obtain more barley mesophyll protoplasts of good quality separated from these barley seedlings.2)The qualities of barley mesophll protoplasts obtained by separating from the barley seedlings cultured 5 days in above culture condition are good,and mainly appeared to obvously increase the percentage of mesophyll protopasts,which could be used to transfer exogenous calcein into them by electroporation , and to have higher calcein transfer efficiency than in ones separated from the seedlings cultured 4 or 6-7 days.The above results provided a usable pre-experimemt or a good model system for using to thorougn studies of gene electee-transfer into barley mesophyll protoplasts > which will be established by using GUS gene.
出处
《杭州大学学报（自然科学版）》
CSCD
1994年第2期220-226,共7页
Journal of Hangzhou University Natural Science Edition
基金
浙江省自然科学基金资助项目
关键词
大麦
叶肉
原生质体
钙黄素
电击法
barley
mesophyll protoplasts
calcein
electroporation
