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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Pyramiding expression of maize genes encoding phosphoenolpyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK) synergistically improve the photosynthetic characteristics of transgenic wheat
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Pyramiding expression of maize genes encoding phosphoenolpyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK) synergistically improve the photosynthetic characteristics of transgenic wheat

机译：编码磷酸烯醇丙酮酸羧化酶（PEPC）和丙酮酸正磷酸二激酶（PPDK）的玉米基因的金字塔表达可协同改善转基因小麦的光合特性

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摘要

Using particle bombardment transformation, we introduced maize pepc cDNA encoding phosphoenolpyruvate carboxylase (PEPC) and ppdk cDNA encoding pyruvate orthophosphate dikinase (PPDK) into the C_3 crop wheat to generate transgenic wheat lines carrying cDNA of pepc (PC lines), ppdk (PK lines) or both (PKC lines). The integration, transcription, and expression of the foreign genes were confirmed by Southern blot, Real-time quantitative reverse transcription PCR (Q-RT–PCR), and Western blot analysis. Q-RT-PCR results indicated that the average relative expression levels of pepc and ppdk in the PKC lines reached 10 and 4.6, respectively, compared to their expressions in untransformed plants (set to 1). The enzyme activities of PEPC and PPDK in the PKC lines were 4.3- and 2.1-fold higher, respectively, than in the untransformed control. The maximum daily net photosynthetic rates of the PKC, PC, and PK lines were enhanced by 26.4, 13.3, and 4.5 %, respectively, whereas the diurnal accumulations of photosynthesis were 21.3, 13.9, and 6.9 %, respectively, higher than in the control. The F_v/F_m of the transgenic plants decreased less than in the control under high temperature and high light conditions (2 weeks after anthesis), suggesting that the transgenic wheat transports more absorbed light energy into a photochemical reaction. The exogenous maize C_4-specific pepc gene was more effective than ppdk at improving the photosynthetic performance and yield characteristics of transgenic wheat, while the two genes showed a synergistic effect when they were transformed into the same genetic background, because the PKC lines exhibited improved photosynthetic and physiological traits.
机译：通过粒子轰击转化，我们将编码磷酸烯醇丙酮酸羧化酶（PEPC）的玉米pepc cDNA和编码丙酮酸正磷酸二激酶（PPDK）的ppdk cDNA引入到C_3作物小麦中，以产生携带pepc（PC系），ppdk（PK系）cDNA的转基因小麦品系或两者兼有（PKC行）。通过Southern印迹，实时定量逆转录PCR（Q-RT–PCR）和Western印迹分析证实了外源基因的整合，转录和表达。 Q-RT-PCR结果表明，与未转化植物（设置为1）中的表达相比，PKC系中pepc和ppdk的平均相对表达水平分别达到10和4.6。与未转化的对照相比，PKC系中PEPC和PPDK的酶活性分别高4.3和2.1倍。 PKC，PC和PK品系的最大每日净光合速率分别提高了26.4％，13.3％和4.5％，而光合作用的日累积量分别比对照组高21.3％，13.9％和6.9％。 。在高温和强光条件下（花后2周），转基因植物的F_v / F_m下降幅度小于对照，这表明转基因小麦将更多吸收的光能传递到光化学反应中。玉米C_4特异的pepc基因在提高转基因小麦的光合性能和产量特性方面比ppdk更有效，而当两个基因转化为相同的遗传背景时，这两个基因表现出协同作用，因为PKC品系表现出改善的光合作用和生理特征。

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