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小麦成熟期穗发芽是世界性的自然灾害,严重影响小麦的产量和品质。Viviparous-1(Vp-1)是促进胚成熟和休眠的主要转录调节因子,与小麦穗发芽抗性有着密切的关系。本实验根据小麦Vp-1基因序列,以植物表达载体pAHC25为基础,成功构建了含有反向重复序列的RNA干扰表达载体pAHC-WVpRi。采用基因枪法轰击小麦品种新春9号幼胚材料1825个,共获得34株T0再生植株。利用Bar基因引物和干扰片段特异引物对再生植株进行PCR检测,获得Bar基因和干扰片段均为阳性的植株3株,转化率为0.16%。本研究为深入分析Vp-1基因功能,进而通过分子育种进行小麦穗发芽抗性的遗传改良提供了科学依据。
Sprouting in mature wheat is a worldwide natural disaster that seriously affects the yield and quality of wheat. Viviparous-1 (Vp-1) is a major transcriptional regulator that promotes embryonic maturation and dormancy, and is closely related to spikelet resistance in wheat. Based on the wheat Vp-1 gene sequence and the plant expression vector pAHC25, we successfully constructed the RNA interference expression vector pAHC-WVpRi with inverted repeat sequences. Using particle bombardment of wheat varieties, 1825 young embryo materials of Chinese New Year 9, a total of 34 T0 regenerated plants were obtained. Using Bar gene primers and interference fragment specific primers for PCR detection of regenerated plants, 3 plants with Bar gene and interference fragment were obtained, and the transformation rate was 0.16%. This study provides a scientific basis for the further analysis of Vp-1 gene function, and then genetic improvement of wheat sprouting resistance by molecular breeding.