摘要
为了获得具有类似C4光合途径的转pepc（磷酸烯醇式丙酮酸羧化酶）基因小麦材料,采用基因枪法将玉米C4型全长pepc基因导入小麦材料01H186-20-24,经在含4 mg.L-1L-PPT（L-phosphinothricin,草丁膦）的培养基上筛选,获得抗性再生植株,经PCR检测获得118株T0代阳性植株;进一步利用PCR对T1代植株进行筛选,T2代在隔离条件下种植于大田,Southern blot、SDS-PAGE检测表明,外源pepc基因在小麦基因组中得以整合和表达。测定了40个转基因T2代植株和受体对照的净光合速率（Pn）、PEPC活性,结果表明,在大田自然条件下,82.5%的转基因小麦的Pn增加,最大提高18.57%,达到了28.1μmol CO2.m-2.s-1,转基因植株中PEPC活性为对照的1.5~1.98倍。
Transformation and high efficiency expression of the C4-specific pepc（phosphoenolpyruvate carboxylase） gene in wheat is an important approach to improve wheat photosynthetic character and biomass yield.In order to obtain PEPC transgenic wheat with similar C4 photosynthetic pathways,recombination binary vector pPC46 carrying maize C4-specific full-length pepc gene was transformed into immature embryos of wheat line 01H186-20-24 by particle bombardment.A total of 118 contemporary transgenic wheat plants（T0） were obtained after selection on medium containing 4 mg·L-1 L-PPT（L-phosphinothricin,Basta） and confirmed by PCR analysis.Further,T1 transgenic wheat were tested by PCR;T2 planted in isolated field were tested by PCR,southern blotting and SDS-PAGE analysis,and showed the foreign pepc gene were integrated into the wheat genome and were expressed.PEPC enzyme activity and net photosynthetic rate（Pn） in 40 T2 transgenic plants and control plants were measured under quarantine areas in field.The results showed that Pn values in 82.5% of transgenic wheat plants increased,with a max increase of 18.57%,to 28.1 μmol CO2· m-2· s-1.Also,the activities of PEPC in leaves of all transgenic plants were as 1.5 to 1.98 times as the non-transgenic plants,indicating that photosynthetic characters of transgenic wheat plants with maize pepc gene were improved.The progress described above provided materials for further study on effects of pepc expression on wheat photosynthesis,biological yield and wheat breeding with high photosynthetic efficiency.
出处
《麦类作物学报》
CAS
CSCD
北大核心
2010年第2期194-197,209,共5页
Journal of Triticeae Crops
基金
国家"973计划"前期专项(2009CB126009)
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(2008ZX08002-003)
河南省杰出人才计划项目(084200410005)
河南省基础与前沿技术研究计划项目(082300430090)
关键词
小麦
玉米C4型pepc基因
转基因
光合速率
PEPC活性
Wheat（Triticum aestivum L.）
Maize（Zea mays） C4-specific pepc gene
Transgenic wheat
Photosynthetic efficiency
PEPC enzyme activity
