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为了寻找大豆抗病新基因,培育大豆新型抗性品种,利用同源克隆的方法从大豆品种科丰1号中分离出1个新的GmRDR1基因,并对其进行序列分析,组织表达、抗逆境胁迫表达分析及该基因的亚细胞定位研究。结果表明:GmRDR1基因位于大豆基因组的第2号染色体,基因全长为3 956 bp,其中ORF为3 378 bp,编码1 125个氨基酸,相对分子量和等电点分别为279.72×103和4.63;GmRDR1含有RDRs家族的保守序列“DLDGD”;该基因在所有被检测组织中均表达,并且在叶中的表达量最高;荧光定量结果发现:在大豆花叶病毒(Soybean mosaic virus,SMV)处理下,GmRDR1在抗病材料科丰1号中的表达量显著高于感病材料南农1138-2。盐及干旱胁迫下,48 h之内,该基因的表达量明显升高,SA诱导条件下该基因在6 h出现了早期响应,冷害处理下24 h表达量出现了突然的升高。GmRDR1基因的亚细胞定位结果表明:该基因所编码的蛋白定位在细胞核里。根据以上结果判定GmRDR1基因参与了大豆对SMV的抗性反应,并且能够强烈响应盐和干旱的胁迫,因此该基因在大豆抗逆分子育种中具有较好的应用价值。
In order to find a new soybean resistance gene and to cultivate new resistant varieties of soybean, a new GmRDR1 gene was isolated from soybean variety Kefeng No.1 by homologous cloning, and its sequence analysis, tissue expression, anti-stress Analysis of stress expression and subcellular localization of the gene. The results showed that the GmRDR1 gene was located on chromosome 2 of the soybean genome with a total length of 3 956 bp. The ORF was 3 378 bp, encoding 1 125 amino acids. The relative molecular weight and isoelectric point were 279.72 × 103 and 4.63, respectively. GmRDR1 Which contained the conserved sequence of “DLDGD” in RDRs family. The gene was expressed in all tested tissues and had the highest expression level in leaves. Fluorescence quantitative results showed that in Soybean mosaic virus (SMV) , The expression level of GmRDR1 in Kefeng 1, a disease-resistant material, was significantly higher than that of susceptible material Nannong 1138-2. Under salt stress and drought stress, the expression level of this gene was significantly increased within 48 h. The expression of this gene appeared early response at 6 h after SA induction, and suddenly increased at 24 h after cold injury. Subcellular localization of GmRDR1 gene indicated that the protein encoded by this gene was located in the nucleus. Based on the above results, it was concluded that GmRDR1 gene is involved in the resistance of soybean to SMV and strongly responds to salt and drought stress. Therefore, it has a good application value in soybean resistance molecular breeding.