Studies on the introduction of C_4 chloroplast into C_3 plant

由于菠菜所属的藜科既有C_3植物，也有C_4植物，因此本研究以菠菜为材料，探讨了将C_4叶绿体引入C_3植物的可行性。虽然人们认为菠菜无菌苗的生产比较困难，但在2%氯化钠溶液中抽真空和搅拌各30分钟的灭菌方法可以减少污染。愈伤组织形成体系的建立是植物原生质体融合或转化的先决条件。菠菜的根和下胚轴比子叶和真叶形成的愈伤组织多。为鉴别外来叶绿体的引入，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酶切菠菜ctDNA后，电泳观察其RFLP模式。

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.