荧光抗体在热带作物小粒种咖啡的C_3/C_4属性鉴别中的应用

用从烟草提纯的 1,5-二磷酸核酮糖 ( Ru BP)羧化酶制备兔抗 Ru BP羧化酶抗体 ,并以异硫氰酸盐荧光素 ( FITC)标记抗体 .采用直接免疫荧光法对典型 C3植物水稻、C4 植物甘蔗和小粒种咖啡等进行了 Ru BP羧化酶的组织化学定位 .结果表明 :C3和 C4 植物叶切片中 Ru BP羧化酶的分布明显不同 ,C3植物的特异荧光位于叶肉细胞 ,C4 植物的特异荧光绝大部分位于维管束鞘细胞 ;小粒种咖啡的特异荧光仅分布在叶肉细胞 .因此认为 ,小粒种咖啡应属 C3植物

Application of immunofluorescence to C3/C4 attribute identification of a tropical crop, Coffea arabica

Rabbit antiserum raised purified ribulose 1,5 bisphosphate carboxylase (RuBPCase) from tobacco was used to locate RuBPCase in leaf blade transection of classical C 3, C 4 plants and Coffea arabica by direct immunofluorescence method. The antibody was labelled by fluoresecin isothiocyanate (FITC). It was discovered that in classical C 4 plant (sugarcane), the specific fluorescence was located almost exclusively in bundle sheath cell chloroplasts, while in C 3 plant (rice), the specific fluorescence was in mesophyllous cell chloroplasts, which proved the difference in RuBPCase location between classical C 4 and C 3 plants. The specific fluorescence of C.arabica was located only in mesophyllous cell chloroplasts, so it was concluded that C.arabica belongs to C 3 plant.