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以T5转hrpZPsta基因大豆JN29-705-15和JL30-187为材料,利用实时荧光定量PCR技术(Quantitative Real Time PCR,qRT-PCR)检测了目标基因在转基因大豆不同组织中的表达量,分别利用下胚轴侵染法和叶面喷施法鉴定疫霉根腐病抗性和灰斑病抗性,并分析目的基因表达量与疫霉根腐病和灰斑病抗性的相关性。结果表明:hrpZPsta基因在大豆的叶、茎、根、籽粒中均有表达,二个株系平均相对表达量分别为8.2/6.1、0.9/0.7、6.5/4.6和0.8/0.7;T5转hrpZPsta基因大豆抗疫霉根腐病和灰斑病能力与野生型相比均有所提高,JN29-705-15对疫霉根腐病抗性从感病提高到中抗,而JL30-187从中抗提高到抗病;hrpZPsta基因在叶中的表达量也与抗灰斑病能力呈正相关,与病情级别呈极显著负相关。试验结果初步证明了外源基因hrp ZPsta在大豆植株中的表达量与受体植株对疫霉根腐病和灰斑病抗性存在一定的相关性。
Using T5 transgenic hrpZPsta soybean JN29-705-15 and JL30-187 as materials, the expression levels of target genes in different tissues of transgenic soybean were detected by using Quantitative Real Time PCR (qRT-PCR) Hypocotyl infection and foliar spray were used to identify the resistance to Phytophthora capsici and Gray leaf spot. The relationship between the expression level of the target gene and Phytophthora root rot and Gray leaf spot resistance was analyzed. The results showed that the hrpZPsta gene was expressed in the leaves, stems, roots and seeds of soybean. The average relative expression levels of the two strains were 8.2 / 6.1, 0.9 / 0.7, 6.5 / 4.6 and 0.8 / 0.7, respectively. The resistance to R. solanacearum and R. solanacearum increased compared with that of wild type. The resistance of JN29-705-15 to P. phyliformis increased from susceptible to moderately resistant, while that of JL30-187 increased from neutral To disease resistance. The expression level of hrpZPsta in leaves was also positively correlated with the ability of resistance to Cercospora sojina and was significantly negatively correlated with the severity of disease. The results showed that the expression level of exogenous gene hrp ZPsta in soybean plants was positively correlated with resistance to Phytophthora capsici and gray leaf spot.