论文部分内容阅读
研究针对从甘菊中克隆获得的DlNAC1基因(Gen Bank登录号为EF602305)进行生物信息学分析,并利用根癌农杆菌介导的叶盘转化法将该基因在烟草中进行过表达研究。结果发现DlAC1蛋白具有较高亲水性,二级结构中占比最高的为无规则卷曲,并具有N糖基化位点等6类潜在的模体结构和典型的由一个扭曲的反平行β片层和α螺旋组成的NAC结构域。将DlAC1基因在烟草中过表达后,通过PCR方法从55株转化植株中鉴定出36株为阳性植株,并且转基因烟草T_0代植株在45℃高温胁迫后,转35S:DlAC1基因阳性植株生长状况良好,而对照植株发生萎蔫,并且转基因植株叶片含水量显著高于对照植株。然而,在4℃低温胁迫后,发现转基因烟草T1代植株没有提高耐低温能力。甘菊DlAC1基因能够提高烟草植株耐高温能力,为今后菊花抗逆育种提供了科学依据。
The bioinformatics analysis of DlNAC1 gene cloned from chamomile (Gen Bank Accession No. EF602305) was carried out and the gene was overexpressed in tobacco by Agrobacterium tumefaciens-mediated leaf disc transformation. The results showed that DlAC1 protein has high hydrophilicity, the highest proportion of secondary structure is randomly curled, and there are 6 types of potential motifs, including N-glycosylation sites, and the typical structure consisting of a twisted anti-parallel β NAC domains composed of lamellar and alpha helix. After the DlAC1 gene was overexpressed in tobacco, 36 plants were identified as positive by PCR from 55 transformed plants. After transgenic plants were exposed to 45 ℃ for 45 days, 35S: DlAC1 positive plants grew well , While the control plants wilted, and the leaf water content of the transgenic plants was significantly higher than that of the control plants. However, after low temperature stress at 4 ℃, T1 plants of transgenic tobacco were found to have no ability to improve low temperature tolerance. The chamomile DlAC1 gene can improve the ability of tobacco plants to resist high temperature and provide a scientific basis for the anti-retrograde breeding of chrysanthemum in the future.