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采用“成熟胚原位转化法”用带有质粒p Cambia2300-Lcchyb-bar的根癌农杆菌C58,侵染玉米芽尖生长点,将chyb基因转入玉米优良品系7922中。T0代植株不进行筛选,T1代植株用浓度为250 mg/L草铵膦筛选,存活植株进行PCR和RT-PCR检测,T1代获得24株阳性植株。T2代植株用500 mg/L草铵膦筛选,PCR阳性植株HPLC分析结果表明转基因植株总类胡萝卜素含量较野生型显著提高,其中玉米黄质含量提高了55.81%。转基因玉米叶片净光合速率明显高于野生型。本研究初步证明目的基因chyb已经成功整合到玉米基因组中,为进一步验证chyb基因在玉米中的功能,以及通过基因工程获得高品质的玉米种质资源奠定基础。
The “in situ transformation of mature embryos” was used to infect corn shoot tip growth sites with Agrobacterium tumefaciens C58 harboring the plasmid pCambia2300-Lcchyb-bar, and the chyb gene was transferred into the maize elite line 7922. T0 plants were not screened, T1 plants were screened with glufosinate at a concentration of 250 mg / L, and the surviving plants were tested by PCR and RT-PCR. Twenty-one positive plants were obtained in T1 generation. The T2 generation plants were screened with 500 mg / L glufosinate. The HPLC analysis of the PCR positive plants showed that the total carotenoid content of the transgenic plants was significantly higher than that of the wild type, with the content of zeaxanthin increased by 55.81%. The net photosynthetic rate of transgenic maize leaves was significantly higher than that of wild type. This study initially proved that the target gene chyb has been successfully integrated into the genome of maize and laid the foundation for further verifying the function of chyb gene in maize and obtaining high quality maize germplasm resources through genetic engineering.