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在克隆获得MYB转录因子GmPHR1基础上,构建基因超表达载体,采用农杆菌介导子叶节转化技术将其转入冀豆12,获得GmPHR1高效表达转基因新材料,并分析基因在耐低磷和低磷敏感品种中的表达差异。结果表明:成功构建了目的基因超表达载体pBI121-GmPHR1,获得了经PCR验证为阳性的T4转基因新材料;荧光实时定量PCR检测发现,其中4个株系的GmPHR1表达量达到野生型对照的2倍以上,说明GmPHR1能够在转基因新材料中高效表达;进一步分析低磷胁迫下不同耐低磷特性品种的基因表达量,发现GmPHR1在耐低磷品种中呈现快速诱导、持续表达与高表达量的模式,而在低磷敏感品种中则表现缓慢诱导、迅速下降和低表达量的模式。
Based on the cloning of the MYB transcription factor GmPHR1, the gene overexpression vector was constructed and transformed into Jidou 12 by Agrobacterium tumefaciens-mediated cotyledonary node transformation to obtain GmPHR1 high efficient transgenic new material. Differences in expression in phosphorus sensitive varieties. The results showed that the target gene overexpression vector pBI121-GmPHR1 was successfully constructed and a new T4 transgenic material verified by PCR was obtained. Real-time fluorescence quantitative PCR showed that the expression of GmPHR1 in four of the four lines reached 2 Times, indicating that GmPHR1 can be highly expressed in new transgenic materials. Further analysis of the gene expression levels of different low-phosphorus tolerant cultivars under low-phosphorus stress showed that GmPHR1 was rapidly induced in low-phosphorus-tolerant cultivars, and was highly expressed Mode, while in low-P-sensitive varieties, it showed a pattern of slow induction, rapid decline and low expression level.