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为深入分析光合作用关键酶二磷酸核酮糖羧化酶(Rubisco)小亚基在油菜等“绿色种子”发育过程中的作用,采用RT-PCR技术从油菜种胚中克隆了一个含Rubisco小亚基全长编码区的cDNA序列,命名为BnrbcS(GenBank登录号DQ242646)。BnrbcS编码181个氨基酸残基,其推导的氨基酸序列中包含典型的Rubisco小亚基功能域并与其他高等植物Rubisco小亚基具有很高的同源性,暗示BnrbcS基因产物与拟南芥等植物的Rubisco小亚基在结构和功能上可能十分相似。除了在叶、子叶、角果皮等光合器官中有很高表达外,BnrbcS在贮藏物质高速积累时期的未成熟油菜种子中亦有中等水平的表达,且其“钟型”表达模式与一些脂肪酸合成酶系基因的表达模式相类似。将NAPIN启动子驱动的BnrbcS种子特异超表达结构转入拟南芥,转化株系的种子含油量和种子重量有一定程度提高,显示BnrbcS在调控种子油脂等贮藏物质积累过程中具有作用。
In order to further analyze the role of the Rubisco subunit, a key enzyme in photosynthesis, in the development of “green seeds” such as rapeseed, a cDNA fragment containing The cDNA sequence of the full-length coding region of the Rubisco small subunit, designated BnrbcS (GenBank Accession No. DQ242646). BnrbcS encodes 181 amino acid residues. The deduced amino acid sequence of BnrbcS contains typical Rubisco small subunit functional domains and has high homology with Rubisco small subunits of other higher plants, suggesting that the BnrbcS gene product interacts with plants such as Arabidopsis The Rubisco small subunit may be very similar in structure and function. In addition to high expression in photosynthetic organs such as leaves, cotyledons, and pericarps, BnrbcS expressed moderately in immature rapeseed seeds during high-speed accumulation of stored matter, and its “bell-shaped” expression pattern was associated with some The expression patterns of fatty acid synthase genes are similar. The results showed that BnrbcS could play an important role in regulating the accumulation of seed oil and other storage materials by transferring the NAPIN promoter-driven BnrbcS seed-specific overexpression structure into Arabidopsis thaliana. The seed oil content and seed weight of the transformed lines increased to some extent.