黃瓜低霜霉威殘留性相關(guān)基因SDH克隆及表達
發(fā)布時間：2018-11-05 07:21
【摘要】：通過Solexa技術(shù)和比較基因組學(xué)方法鑒定霜霉威脅迫下黃瓜果實差異表達基因,篩選黃瓜低霜霉威殘留性相關(guān)基因SDH。研究利用PCR克隆獲得SDH基因全長,命名為Cs SDH。構(gòu)建亞細(xì)胞定位表達載體,利用農(nóng)桿菌浸潤法注射煙草葉片后激光共聚焦顯微鏡下觀察;分別用熒光定量RT-PCR方法和酶聯(lián)免疫法研究高、低農(nóng)殘品系D9320和D0351中Cs SDH基因在農(nóng)藥霜霉威處理后,不同時間點、不同組織部位時空表達特征和琥珀酸脫氫酶活性。結(jié)果表明,Cs SDH基因在煙草葉片細(xì)胞中被定位在細(xì)胞膜上。黃瓜低農(nóng)殘品種D0351中,Cs SDH基因在果實受霜霉威脅迫后反應(yīng)迅速,主要在處理前期響應(yīng)表達強烈,琥珀酸脫氫酶活性顯著增加,且該基因表達量顯著高于其在高農(nóng)殘品種D9320中表達量。Cs SDH果、葉中表達量較莖中高,且各組織部位表達量高低順序穩(wěn)定,為葉果莖。黃瓜高農(nóng)殘品種D9320中,Cs SDH基因表達量在0.5、1、3、12 h呈現(xiàn)上調(diào)表達,6、9 h基因表達量差異不顯著,48 h出現(xiàn)上調(diào)表達高峰。說明克隆得到的Cs SDH基因?qū)俎r(nóng)藥處理前期響應(yīng)基因,且該基因可能在降低黃瓜農(nóng)藥殘留過程中發(fā)揮重要作用。試驗通過研究黃瓜琥珀酸脫氫酶基因(SDH)在霜霉威脅迫下表達情況,為挖掘黃瓜低農(nóng)藥殘留性功能基因以及探明其作用分子機制奠定基礎(chǔ)。
[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.
【作者單位】： 東北農(nóng)業(yè)大學(xué)園藝學(xué)院;
【基金】：國家自然科學(xué)基金項目(31272158)
【分類號】：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.
【作者單位】： 東北農(nóng)業(yè)大學(xué)園藝學(xué)院;
【基金】：國家自然科學(xué)基金項目(31272158)
【分類號】：S642.2;S481.8
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