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水稻NRT1.1B基因是已经克隆并进行功能验证的氮高效利用基因,具有很高的应用价值。根据籼粳稻NRT1.1B基因的基因组序列比对发现,籼型NRT1.1B基因及粳型NRT1.1B基因内含子序列中存在插入/缺失位点。对30个常规籼粳稻的NRT1.1B基因内含子序列进行PCR扩增、测序及序列比对,发现NRT1.1B基因内含子存在6个In Del位点,籼型NRT1.1B基因比粳型NRT1.1B基因缺失55 bp。根据插入/缺失位点设计出In Del分子标记,对15个籼稻常规稻品种、15个粳稻常规稻品种、3个籼粳杂交稻品种及20个F2代育种材料进行NRT1.1B籼粳基因型鉴定。检测实验表明与通过NRT1.1B基因的SNP位点开发的功能标记的检测完全一致。通过该标记可以准确鉴定NRT1.1B基因的纯合籼型、纯合粳型及杂合基因型,该方法成本低、简单、可靠,可用于NRT1.1B基因的鉴定和分子标记辅助育种。
The rice NRT1.1B gene is a nitrogen efficient utilization gene that has been cloned and verified by function, and has high application value. According to the genome sequence alignment of indica and japonica NRT1.1B genes, it is found that there are insertion / deletion sites in intron sequences of indica NRT1.1B gene and japonica NRT1.1B gene. Intron 3 of NRT1.1B gene of 30 indica and japonica rice was amplified by PCR, sequenced and sequenced. It was found that there were 6 In Del sites in intron of NRT1.1B gene. The indica type NRT1.1B gene was more than japonica Type NRT1.1B gene deletion of 55 bp. The In Del molecular markers were designed according to the insertion / deletion loci. The indica-japonica genotypes of 15 indica rice varieties, 15 japonica conventional rice varieties, 3 indica japonica hybrid rice varieties and 20 F2 breeding materials were identified. Identification. The test shows that the detection of the functional marker developed by the SNP site of NRT1.1B gene is exactly the same. The marker can accurately identify the homozygous indica, homozygous japonica and heterozygous genotypes of NRT1.1B gene. The method is low cost, simple and reliable, and can be used for identification of NRT1.1B gene and molecular marker-assisted breeding.