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作物学报 ›› 2015, Vol. 41 ›› Issue (03): 507-514.

• 研究简报 • 上一篇    

稗草(Echinochloa crusgalli)根型ppc 基因对水稻的遗传转化及其对光合速率的调节效应

张桂芳1,2,丁在松1,赵明1,*   

  1. 1 中国农业科学院作物科学研究所 / 农业部作物生理生态重点实验室, 北京 100081; 2 北京师范大学生命科学学院 / 北京师范大学学报(自然科学版)编辑部, 北京 100875
  • 收稿日期:2014-05-09 修回日期:2014-12-19 出版日期:2015-03-12 网络出版日期:2015-01-28
  • 通讯作者: 赵明, E-mail: zhaomingcau@vip.tom.com
  • 基金资助:

    本研究由中国农业科学院创新工程项目资助。

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,*   

  1. 1 Institute of Crop Science, Chinese Academy of Agricultural Sciences / Key Laboratory of the Ministry of Agriculture Crop Physiology and Ecology,
    Beijing 100081,China; 2 College of Life Science / Editorial Department of Journal of Beijing Normal University (Natural Science), Beijing 100875, China
  • Received:2014-05-09 Revised:2014-12-19 Published:2015-03-12 Published online:2015-01-28
  • Contact: 赵明, E-mail: zhaomingcau@vip.tom.com

摘要:

稗草(Echinochloa crusgalli)是稻田中的C4 光合型杂草, 为了探索稗草ppc 基因(Eppc)对水稻遗传转化的可行性及其对光合速率的调节效应, 首次将含有稗草根型磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase, PEPC)基因ppc cDNA 的2 个植物表达载体pUbi-Eppc、pRbcS-Eppc 通过农杆菌介导法对水稻进行了遗传转化。对分化植株进行的PCR、RT-PCR、克隆测序和Western 杂交等结果均表明稗草ppc 基因已经整合到了水稻基因组中, 并且在转录和翻译水平都得到了表达。转基因水稻PEPC 活性和气体交换参数测定结果表明T0 代多数植株的PEPC 活性高于对照, 最高达到了对照的5.85 倍; T0 代大多数转基因植株叶片的净光合速率(Pn)比对照提高了20.00%, 最大地提高了47.16%, 同时叶片水分利用效率(WUE)也得到增强; T6 代大部分转化植株的PEPC 活性及Pn 仍保持高于对照, 本研究表明C3 根型ppc 基因过量表达也可以提高水稻的Pn, 且证明稗草PEPC 对光合作用具有较强的调节作用。

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

Abstract:

Barnyardgrass (Echinochloa crusgalli) is a C4 weed commonly found in rice field. To fully utilize the photosynthestic potential of Barnyardgrass C4 gene, 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.

Key words: Echinochloa crusgalli, Root phosphoenolpyruvate carboxylase gene, Rice, PEPC activities, Net photosynthetic efficiency, Water use efficiency

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