Studies on the introduction of C_4 chloroplast into C_3 plant

本研究以菠菜为材料，探讨了将C_4叶绿体导入C_3植物的可行性。菠菜属藜科植物C_3和C_4植物同属一科。虽然菠菜很难产生无菌苗，但用2%NaClO_2抽真空搅拌种子30分钟可以减少污染。建立愈伤组织形成体系是植物原生质体融合或转化的前提。菠菜根和下胚轴切段的愈伤组织形成率高于子叶和真叶切段。为了鉴定外源叶绿体的导入，对叶绿体DNA分析技术进行了研究。用限制性内切酶BamHI、EcoRI、PstI和ApaI对菠菜ctDNA进行限制性内切酶消化后，在凝胶上观察到RFLP图谱。

In the present study, the feasibility of the introduction of C_4 chloroplast into C_3 plant was examined using spinach because Chenopodiaceae to which spinach belongs has both C_3 and C_4 plants in the same family.Though spinach had been considered to be difficult in producing the aseptic seedlings, the sterilization of seeds by vacuumming and stirring them for 30 minuts each in 2% NaClO_2 could reduce the contamination.To establish the callus formation system is a prerequisite for the protoplast fusion or transformation of plants. More callus formation was obtained from root and hypocotyl segments of spinach than from cotyledon and true leaf segments.To identify the introduction of alien chloroplasts, the technique of analysis of chloroplast DNA was investigated. RFLP patterns of ctDNA of spinach were observed at the electrophoresis after digested by the restriction enzyme, BamHI,EcoRI,PstI and ApaI.