Studies on the introduction of C_4 chloroplast into C_3 plant

C_4叶绿体导入C_3植物的研究

• 批准号: 06806003
• 负责人: TANAKA Takayuki
• 金额: $ 0.7万
• 依托单位: Kyushu Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06806003/
• 关键词: spinach; aseptic sowing; callus; chloroplast; DNA; プロトプラスト; サイトプラスト; 高張液; カリオプラスト

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.

本研究利用菠菜检验将C_4叶绿体引入C_3植物的可行性，因为菠菜所属的藜科植物同时具有C_3和C_4植物，虽然菠菜一直被认为很难生产无菌苗种子在2% NaClO_2中真空搅拌30分钟进行灭菌,可减少污染。愈伤组织形成体系的建立是愈伤组织形成的前提。植物的原生质体融合或转化。与子叶和真叶片段相比，从菠菜的根和下胚轴片段获得更多的愈伤组织形成。为了鉴定外来叶绿体的引入，研究了叶绿体DNA分析技术。菠菜ctDNA经限制性内切酶BamHI、EcoRI、PstI和ApaI酶切后电泳观察RFLP图谱。