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。
To develop new transgenic soybean materials with MYB transcription factor GmPHR1, and to study the expression patterns of GmPHR1 in different low-P tolerant soybean varieties, a over-expression vector pBI121-GmPHR1 was constructed and introduced into Jidoul2 by Agrobacterium-mediated cotyledonary-node transformation method. PCR and real-time quantita- tive PCR analysis results of the transformed T4plants showed that the GmPHR1 was successfully incorporated into the soybean genome and expressed at a higher level than the wild-type. Expression patterns assayed by real-time quantitative PCR revealed that the expression level of GmPHR1 in low-P tolerant variety NF37 was induced quickly for a longer period at a higher level than that in sensitive variety Niumaohuang under phosphate starvation condition.