离体培养植物细胞的表面与农杆菌介导基因转化频率的研究

Relationship Between the Outer Surfaces of Plant Cells Cultured in vitro and the Frequencies of Transformation Agrobacterium mediated

以玉米、黑麦、烟草为材料 ,对细胞表面进行了扫描电镜观察 ,研究了离体培养植物细胞的细胞壁外表面的结构及其变化与农杆菌介导转化频率之间的关系。正常继代培养的烟草细胞 ,其细胞壁外表面光滑 ,大部分细胞在扫描电镜下坍陷或皱折 ;经处理用于农杆菌介导转化的烟草细胞 ,其表面有很多颗粒状突起和纤丝状物或纤丝状网 ;光滑的表面很少见到菌 ,而粗糙表面则有很多菌。玉米和黑麦愈伤组织中 ,浅黄色、疏松易散的胚性细胞团 ,其表面细胞有很多未坍陷 ,相当一部分未坍陷细胞的细胞壁外表面与烟草细胞一样 ,具有很多较小的颗粒状突起和纤丝状物或纤丝状网 ;白色松软的愈伤组织 ,其外层细胞大多坍陷或呈柱状 ,未坍陷细胞的表面光滑 ,有些似有一层膜状物。

The outer surfaces of plant cells from tobacco (Nicotiana tabacum),maize( Zea mays ),and rye( Secale cereale )cultured in vitro have been studied using scanning electron microscopy(SEM).The outer surfaces of cell wall(OSCW) of the cells from tobacco subcultures are smooth,and most of the cells are collapsed or crinkled after specimen prepared for SEM.In contrast,there are many protuberances or beads and/or fibres or meshwork on OSCW of the tobacco cells treated for Agrobacterium mediated transformation.Bacteria crowd on the granulated surfaces of the tobacco cells,but little on the smooth surfaces.The same as tobacco cells,OSCW are granulated for the cells from pale yellow,soft and friable and fast grown embryogenic calluses of rye and from the pale yellow calluses of maize.OSCW are smooth for the cells from white and soft calluses of maize,and only a few cells haven't been collapsed in these calluses.The relationship between Agrobacterium attachment sites,which could be induced,on the cell wall surfaces,and the frequencies of genetic transformation mediated by Agrobacterium were discussed.