关键词:稗草; 根型 Eppc基因; 水稻; PEPC活性; 净光合速率; 水分利用效率 Transformation of Barnyardgrass ( Echinochloa crusgalli) Root Type Phosphoenolpyruvate Carboxylase Gene into Rice ( Oryza sativa) Plants and Their Effects on Photosynthesitic Gas Exchange ZHANG Gui-Fang1,2, DING Zai-Song1, ZHAO Ming1,* 1Institute of Crop Science, Chinese Academy of Agricultural Sciences / Key Laboratory of the Ministry of Agriculture Crop Physiology and Ecology, Beijing 100081, China
2College of Life Science / Editorial Department of Journal of Beijing Normal University (Natural Science), Beijing 100875, China
AbstractBarnyardgrass ( Echinochloa crusgalli) is a C4 weed commonly found in rice field. To fully utilize the photosynthestic potential of Barnyardgrass C4gene, we transformed Barnyardgrass root Phosphoenolpyruvate Carboxylase gene into rice plant with vectors contained promoters of Ubiqitin gene and Rubisco small unit gene by Agrobactirium- mediated transformation. Both marker genes Hygr and ppc were detected by PCR in regenerated plants. RT-PCR and Western blot analysis confirmed that the ppc gene was incorporated into rice plant and expressed with stable transcripts and proteins. PEPC activity as measured in most of the transgenic rice plants was higher than that in control, being up to 5.85-fold of that in untransformed rice. At T0 generation, net photosynthetic rate ( Pn) in most of transgenic rice plants was 20.00% higher than that in untransformed rice, with the highest increase of 47.16%. Water utilization efficiency (WUE) in transgenic rice was also improved. At T6 generation, PEPC activity and Pn of transgenic lines remained higher than those of the wild type. These indicate that over-expressing C3 Eppc gene also can improve rice photosynthesis.
Keyword:Echinochloa crusgalli; Root phosphoenolpyruvate carboxylase gene; Rice; PEPC activities; Net photosynthetic efficiency; Water use efficiency Show Figures Show Figures
图2 筛选标记基因Hygr的PCR检测 泳道1~7为pUbi-Eppc载体的转基因植株, 8~14为pRbcS-Eppc载体的转基因植株; M为标准DNA长度, 依次为300、500、1000、1500、2000和2500 bp, WT为未转化的品种中花8号。Fig. 2 PCR detection of Hyg resistance gene in transgenic rice Lane 1-7: Transformants of vector pUbi-Eppc; Lane 8-14: Transformants of vector pRbcS-Eppc; M: DNA marker, being 300, 500, 1000, 1500, 2000, 2500 bp in length; WT: untransformed Zhonghua 8.
图3 转基因水稻中Eppc基因的PCR扩增检测 泳道1~12为pRbcs-Eppc转基因水稻样本扩增产物, WT为对照, M为DNA标准长度, 依次为500、1000、2500、5000、7500 bp)Fig. 3 PCR detection of Eppc gene in transgenic rice Lane 1-12: Transformants of vector pRbcS-Eppc; M: DNA marker, being 500, 1000, 2500, 5000, 7500 bp in length; WT: untransformed Zhonghua 8.
图5 转基因水稻PEPC蛋白的Western检测 泳道1~4和5~8分别为pUbi-Eppc和pRbcS-Eppc载体的转基因植株, WT为对照中花8号。Fig. 5 Western analysis of PEPC in transgenic rice plants Lane 1-4: transformants of vector pUbi-Eppc; Lane 5-8: transformants of vector pRbcS-Eppc; WT: untransformed Zhonghua 8.
图6 转稗草ppc基因植株的PEPC相对活性 uZH8-n: pUbi-Eppc的转化苗; rZH8-n: pRbcS-Eppc的转化苗; WT: 中花8号。Fig. 6 Relative PEPC activity improved by transgenic of Eppc in the control of different promoters uZH8-n: transformants of vector pUbi-Eppc; rZH8-n: transformants of vector pRbcS-Eppc; WT: untransformed Zhonghua 8.
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