| 张建红，许为钢，王会伟，李 艳，胡 琳，韩琳琳，张会芳.转玉米C4型高光效pepc基因小麦的分子特征及光合特性研究[J].麦类作物学报,2012,32(6):1043 |
| 转玉米C4型高光效pepc基因小麦的分子特征及光合特性研究 |
| Molecular Characteristics and Photosynthetic Property of the Transgenic Wheat Expressing a Maize C4-type pepc Gene |
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| DOI：10.7606/j.issn.1009-1041.2012.06.006 |
| 中文关键词: 小麦 pepc基因 表达 净光合速率 |
| 英文关键词:Wheat Transgenic pepc Net photosynthetic rate |
| 基金项目:国家自然科学基金项目(30971785)；国家转基因生物新品种培育科技重大专项(2011ZX08002-003)；国家科技支撑计划项目(2011BAD07B01)；现代农业产业技术体系专项(CARS-3-1-9)。 |
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| 中文摘要: |
| 为分析转pepc基因小麦的分子遗传特征和光合生理特性，以基因枪介导转玉米C4型pepc基因的T3代转基因小麦株系为实验材料，对其进行PCR筛选、Southern 杂交、qRT-PCR、Western 杂交、酶活性和光合速率测定，鉴定了pepc基因在小麦基因组中的整合及表达情况，并分析了转基因株系的光合生理特性和产量性状表现。结果表明，玉米C4型pepc基因以1~3个拷贝的方式被整合到6个转基因株系的不同位点，且可以正确转录和翻译；6个转基因株系中有4个株系的pepc基因表达水平、PEPC酶活性和净光合速率均较对照显著提高，2个株系的单株重、单株产量和千粒重显著高于对照；抽穗期转基因株系的PEPC酶活性最大值为1.6 μmol·min-1·mg-1，较对照提高了1.6倍；净光合速率最大值为29.5 μmol·m-2·s-1，较对照提高了15.5%。说明，玉米C4型pepc基因可以在小麦基因组中整合表达，并可改善小麦的光合特性。 |
| 英文摘要: |
| The objective of this study is to analyze the molecular characteristics and physiological property of the transgenic wheat expressing a maize C4-type pepc gene. Using the particle bombardment transformation method, PEPC, a key C4 enzyme gene from maize, was transferred into wheat. The PEPC integration and expression in the T3 PEPC transgenic lines were analyzed using Southern and Western blots, and qRT-PCR. Enzyme activity and photosynthetic rates were also evaluated in the PEPC- transgenic lines. One to three PEPC copies were identified by Southern blot analysis in the 6 PEPC transgenic lines and the transcripts and protein products of PEPC were detected by qRT-PCR and Western blot analyses. These indicated that the pepc gene had steadily integrated and expressed well in the wheat genome. Enzyme activity and photosynthetic rate determinations showed that 4 PEPC transgenic lines had higher PEPCase activities and net photosynthesis rate than controls, which had the peak of 1.6 μmol·min-1·mg-1 (up-1.6 fold of controls) and 29.5 μmol·m-2·s-1 (up-1.16 fold of controls), respectively. The C4-type pepc gene from maize could be integrated and expressed in the wheat genome and improve the photosynthetic characteristics of wheat. This study will provide supporting materials for high photosynthesis wheat breeding. |
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