摘要
采用电脉冲法，转化大分子复合物ＩｇＧ－荧光素进入小麦部分酶解（１２０ｍｉｎ）愈伤组织的细胞即半原生质体中，在荧光显微镜下观察发现一些细胞内有荧光发生，由此确定的较适宜脉冲转化条件如电场强度为２２５０－３０５０Ｖ／ｃｍ应用于转化外源基因ｐＺＦＸ２到小麦半原生质体，随后将其按愈伤组织的培养过程进行培养和抗ｋａｎａｍｙｃｉｎ筛选后获得成活愈伤细胞团块，存活率为１１．３％－１８．９％，这都说明已成功地将这两种物质转入了小麦细胞内，从而在小麦中可以绕过原生质体。
After introduction of IgG FITC compound into semiprotoplast by electroporation,fluorescences in some semiprotoplast cells was observed in Opton Fluorescence Microscope.Through this test,we obtained the better conditions of electroporation.For example,the electric pulse field was 2250-3050 V/cm.Then plasmid pZFX2,containing the NPT Ⅱ (neomycin phosphotransferase) and glycolate oxidase gene which can in crease the wheat plant resistance to insects,was introduced into semi protoplast.Some transgenic semi protoplast cells were cultured on MS medium containing 100 μg/mL kanamycin.After 30 days election culture,11.3%-18.9% of the semi protoplast cell clumps survived.The results proved that the semi protoplast cell clumps may be potential,feasible receptor for gene transformation in wheat,especially in those varieties or lines which are difficult for protoplast culture.
出处
《西北植物学报》
CAS
CSCD
1999年第1期29-34,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
本研究为云南省自然科学基金
关键词
基因转化
NPT-Ⅱ
半原生质体
小麦
gene transformation,IgG FITC,NPT Ⅱ,semi protoplast,wheat
