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OsHOX4基因是属于水稻植物同源结构域(HD-Zip)转录因子家族的一个成员。本研究利用农杆菌介导法将OsHOX4基因转入籼稻(Oryza sativa ssp.indica)IR64中,获得54株过量表达植株。我们利用southern杂交分析确定了各个株系的拷贝数后,利用TAIL-PCR方法对4个不同的单拷贝株系进行了研究,并在其中找出T-DNA在3个单拷贝株系染色体上的插入位点(分别为染色体5,8,10)。根据T-DNA在各个株系的插入位点分别设计特异性引物,从T1代转基因株系里鉴定出纯合的植株。我们观察到OsHOX4过量表达植株的分蘖数明显高于野生型,株高比野生型明显的变矮,在同一个株系的纯合和杂合的转基因植株表型也有很大的差异。在TAIL-PCR分析基础上,为从T1代转基因作物中鉴定出纯合的植株提供了方法和理论依据。
The OsHOX4 gene is a member of the HD-Zip transcription factor family of rice plants. In this study, OsHOX4 gene was transferred into Oryza sativa ssp. Indica IR64 using Agrobacterium-mediated method and 54 overexpression plants were obtained. After confirming the copy number of each line by southern hybridization analysis, we studied four different single-copy lines by TAIL-PCR and found out that T-DNA was on chromosomes of three single-copy lines The insertion sites (chromosomes 5, 8 and 10, respectively). According to the T-DNA design of specific primers at each insertion site, homozygous plants were identified from the T1 generation transgenic lines. We observed that OsHOX4 overexpression plants had significantly higher tiller numbers than wild type plants, significantly shorter plant height than wild type plants, and significant differences in the phenotypes of homozygous and heterozygous transgenic plants in the same line. Based on the TAIL-PCR analysis, the method and theoretical basis for the identification of homozygous plants from T1 transgenic plants are provided.