CAJ | 학술논문

为了获得具有类似C_4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C_4型全长pepc基因导入小麦材料01H186-20-24,经在含4 mg·L~(-1) L-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T_0代阳性植株;进一步利用PCR对T_1代植株进行筛选,T_2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28.1 μmol CO_2· m~(-2)·s~(-1),转基因植株中PEPC活性为对照的1.5～1.98倍.
위료획득구유유사C_4광합도경적전pepc(린산희순식병동산최화매)기인소맥재료,채용기인창법장옥미C_4형전장pepc기인도입소맥재료01H186-20-24,경재함4 mg·L~(-1) L-PPT(L-phosphinothricin,초정련)적배양기상사선,획득항성재생식주,경PCR검측획득 118주T_0대양성식주;진일보이용PCR대T_1대식주진행사선,T_2대재격리조건하충식우대전,Southern blot、SDS-PAGE검측표명,외원pepc기인재소맥기인조중득이정합화표체.측정료40개전기인T2대식주화수체대조적정광합속솔(Pn)、PEPC활성,결과표명,재대전자연조건하,82.5%적전기인소맥적Pn증가,최대제고18.57%,체도료28.1 μmol CO_2· m~(-2)·s~(-1),전기인식주중PEPC활성위대조적1.5～1.98배.
Transformation and high efficiency expression of the C_4-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 C_4 photosynthetic pathways, recombination binary vector pPC46 carrying maize C_4-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 (T_0) were obtained after selection on medium containing 4 mg·L~(-1) L-PPT (L-phosphinothricin, Basta) and confirmed by PCR analysis. Further, T_1 transgenic wheat were tested by PCR;T_2 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 T_2 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 CO_2· 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.