【摘要】：該研究采用RT-PCR技術(shù),從抗病的中國野生華東葡萄‘白河-35-1’和感病的歐洲葡萄‘佳麗釀’中克隆了液泡加工酶基因(γVPE),分別命名為VpγVPE和VvCγVPE。克隆的2個γVPE基因cDNA長度均為1 624bp,ORF為1 482bp,編碼493個氨基酸。氨基酸多序列對比分析發(fā)現(xiàn),‘白河-35-1’、‘佳麗釀’、‘無核白’和‘黑比諾’葡萄中的γVPE基因底物結(jié)合口袋域的3個關(guān)鍵氨基酸之一的絲氨酸(Ser395)均變?yōu)楸彼?Ala),與其他植物的VPE基因底物口袋結(jié)合域有所不同。實時熒光定量PCR表明,在白粉菌誘導(dǎo)后的不同時期內(nèi),γVPE基因在感病葡萄和抗病葡萄中的表達(dá)模式不同,抗病株系中VpγVPE基因的表達(dá)量在誘導(dǎo)后的前期(4h和48h)和后期(168h)均有所增加,而感病株系中VvCγVPE基因在誘導(dǎo)后4h表達(dá)量最高,隨后降低。γVPE基因在白粉菌誘導(dǎo)后不同時期內(nèi)表達(dá)量的變化,表明γVPE基因在一定程度上與葡萄的抗性相關(guān)。研究結(jié)果為進(jìn)一步揭示γVPE基因在抗病過程中的分子機(jī)理奠定了基礎(chǔ)。
[Abstract]:Using RT-PCR technique, the vacuolar processing enzyme gene (緯 VPE),) was cloned from the disease-resistant wild Chinese grape 'Baihe-35-1' and the susceptible European grape 'Jiali'. 緯 VPE), was named Vp 緯 VPE and VvC 緯 VPE., respectively. The cDNA length of the two 緯 VPE genes was 1 624 BP, and the ORF was 1 482 BP, encoding 493 amino acids. Comparative analysis of amino acid sequences revealed that 'Baihe-35-1', 'Jiali', The serine (Ser395), one of the three key amino acids in the substrate binding pocket domain of 緯 VPE gene in 'denuclearized white' and 'Pinot Noir' grapes, is different from the VPE gene substrate pocket binding domain in other plants. Real-time fluorescence quantitative PCR showed that the expression patterns of 緯 VPE gene in susceptible and disease-resistant grapes were different at different stages after induction by powdery mildew. The expression of Vp 緯 VPE gene increased in the early stage (4 h and 48 h) and later stage (168 h) of resistant lines, but the expression of VvC 緯 VPE gene was the highest in susceptible lines at 4 h after induction. The changes of 緯 VPE gene expression in different period after induced by powdery mildew indicated that 緯 VPE gene was related to grape resistance to some extent. The results laid a foundation for further revealing the molecular mechanism of 緯 VPE gene in the process of disease resistance.
【作者單位】： 西北農(nóng)林科技大學(xué)園藝學(xué)院旱區(qū)作物逆境生物學(xué)國家重點實驗室農(nóng)業(yè)部西北地區(qū)園藝作物生物學(xué)與種質(zhì)創(chuàng)制重點實驗室;
【基金】：國家自然科學(xué)基金(31171926) 國家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專項(CARS-30-yz-7) 楊凌示范區(qū)農(nóng)業(yè)科技示范推廣項目(2015-TS-10)
【分類號】：S436.631
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本文編號：2386285