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利用农杆菌介导法将番茄红素β-环化酶基因(Lycb)转入由玉米自交系天塔五号植株,分析基因在T0转化及后代的遗传情况,结果表明,在27株T0转基因植株中,PCR初步检测后8株呈阳性;将T1代转基因植株以株系为单位用200mg/L草铵膦抗性筛选后,收获抗性植株种子。T2代转基因植株进一步进行PCR、RT-PCR和田间草铵膦涂抹检测,结果表明,PCR、RT-PCR为阳性的6个株系植株均具有草铵膦抗性。选取6株阳性植株提取叶片总类胡萝卜素,经HPLC分析其β-胡萝卜素含量显著高于野生型,表明目的基因Lycb成功的转入玉米,并得到了稳定遗传。
The lycopene β-cyclase gene (Lycb) was transferred into Tianta 5, a maize inbred line, by Agrobacterium tumefaciens-mediated method. The genetic variation of T0 gene and its offspring was analyzed. The results showed that in T0 Among the transgenic plants, 8 were positive after the preliminary PCR test. The T1 transgenic plants were screened with 200 mg / L glufosinate-resistant strain and the resistant plants were harvested. The T2 generation transgenic plants were further tested by PCR, RT-PCR and field glufosinate-smear. The results showed that all the six plants tested positive by PCR and RT-PCR had glufosinate resistance. Total 6 carotenoids were extracted from 6 positive plants. The content of β-carotene in the leaves was significantly higher than that in the wild type by HPLC, indicating that the target gene Lycb was transferred into maize successfully and was stable and inherited.