黄瓜低霜霉威残留性相关基因SDH克隆及表达
发布时间：2018-11-05 07:21
【摘要】：通过Solexa技术和比较基因组学方法鉴定霜霉威胁迫下黄瓜果实差异表达基因,筛选黄瓜低霜霉威残留性相关基因SDH。研究利用PCR克隆获得SDH基因全长,命名为Cs SDH。构建亚细胞定位表达载体,利用农杆菌浸润法注射烟草叶片后激光共聚焦显微镜下观察;分别用荧光定量RT-PCR方法和酶联免疫法研究高、低农残品系D9320和D0351中Cs SDH基因在农药霜霉威处理后,不同时间点、不同组织部位时空表达特征和琥珀酸脱氢酶活性。结果表明,Cs SDH基因在烟草叶片细胞中被定位在细胞膜上。黄瓜低农残品种D0351中,Cs SDH基因在果实受霜霉威胁迫后反应迅速,主要在处理前期响应表达强烈,琥珀酸脱氢酶活性显著增加,且该基因表达量显著高于其在高农残品种D9320中表达量。Cs SDH果、叶中表达量较茎中高,且各组织部位表达量高低顺序稳定,为叶果茎。黄瓜高农残品种D9320中,Cs SDH基因表达量在0.5、1、3、12 h呈现上调表达,6、9 h基因表达量差异不显著,48 h出现上调表达高峰。说明克隆得到的Cs SDH基因属农药处理前期响应基因,且该基因可能在降低黄瓜农药残留过程中发挥重要作用。试验通过研究黄瓜琥珀酸脱氢酶基因(SDH)在霜霉威胁迫下表达情况,为挖掘黄瓜低农药残留性功能基因以及探明其作用分子机制奠定基础。
[Abstract]:The differentially expressed genes were identified by Solexa and comparative genomics methods in cucumber fruit under creamethanil stress, and SDH. genes related to the residue of hypocremazine in cucumber were screened. The full length of SDH gene, named Cs SDH., was obtained by PCR cloning. The subcellular expression vector was constructed and observed by laser confocal microscope after the tobacco leaf was injected with Agrobacterium tumefaciens (Agrobacterium tumefaciens). Fluorescence quantitative RT-PCR and enzyme-linked immunosorbent assay (Elisa) were used to study the expression of Cs SDH gene and the activity of succinic dehydrogenase at different time points and different tissue sites in D9320 and D0351 lines. The results showed that, Cs SDH gene was located on the cell membrane in tobacco leaf cells. The response of, Cs SDH gene in cucumber low agricultural residuals D0351 was rapid, mainly in the early stage of treatment, and the activity of succinic dehydrogenase increased significantly. The expression level of this gene was significantly higher than that of. Cs SDH in D9320, and the expression in leaf was higher than that in stem, and the order of expression in different tissues was stable, which was the stem of leaf and fruit. The expression of, Cs SDH gene in D9320, a cucumber cultivar with high agriculture residue, was up-regulated at 12 h after 0. 5 and 1: 1, and showed no significant difference at 6 ~ 9 h, but reached a peak at 48 h. The results indicated that the cloned Cs SDH gene was a response gene in the early stage of pesticide treatment, and the gene might play an important role in reducing pesticide residues in cucumber. The expression of succinic dehydrogenase (SDH) gene in cucumber was studied in order to explore the functional genes of low pesticide residues in cucumber and to explore its molecular mechanism.
【作者单位】： 东北农业大学园艺学院;
【基金】：国家自然科学基金项目(31272158)
【分类号】：S642.2;S481.8
[Abstract]:The differentially expressed genes were identified by Solexa and comparative genomics methods in cucumber fruit under creamethanil stress, and SDH. genes related to the residue of hypocremazine in cucumber were screened. The full length of SDH gene, named Cs SDH., was obtained by PCR cloning. The subcellular expression vector was constructed and observed by laser confocal microscope after the tobacco leaf was injected with Agrobacterium tumefaciens (Agrobacterium tumefaciens). Fluorescence quantitative RT-PCR and enzyme-linked immunosorbent assay (Elisa) were used to study the expression of Cs SDH gene and the activity of succinic dehydrogenase at different time points and different tissue sites in D9320 and D0351 lines. The results showed that, Cs SDH gene was located on the cell membrane in tobacco leaf cells. The response of, Cs SDH gene in cucumber low agricultural residuals D0351 was rapid, mainly in the early stage of treatment, and the activity of succinic dehydrogenase increased significantly. The expression level of this gene was significantly higher than that of. Cs SDH in D9320, and the expression in leaf was higher than that in stem, and the order of expression in different tissues was stable, which was the stem of leaf and fruit. The expression of, Cs SDH gene in D9320, a cucumber cultivar with high agriculture residue, was up-regulated at 12 h after 0. 5 and 1: 1, and showed no significant difference at 6 ~ 9 h, but reached a peak at 48 h. The results indicated that the cloned Cs SDH gene was a response gene in the early stage of pesticide treatment, and the gene might play an important role in reducing pesticide residues in cucumber. The expression of succinic dehydrogenase (SDH) gene in cucumber was studied in order to explore the functional genes of low pesticide residues in cucumber and to explore its molecular mechanism.
【作者单位】： 东北农业大学园艺学院;
【基金】：国家自然科学基金项目(31272158)
【分类号】：S642.2;S481.8
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