Functional diversification in the Nudix hydrolase gene family drives sesquiterpene biosynthesis in Rosa x wichurana
Functional diversification in the Nudix hydrolase gene family drives sesquiterpene biosynthesis in Rosa x wichurana
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DOI：
10.1111/tpj.14916
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发表时间：
2020-07-22
期刊：
影响因子：
7.2
通讯作者：
Baudino, Sylvie
中科院分区：
文献类型：
--
作者：
Sun, Pulu;Degut, Clement;Baudino, Sylvie
玫瑰使用一种非经典的途径，涉及一种NUDX 1水解酶，RhNUDX 1，来合成它们的单萜，特别是香叶醇。在这里，我们报告的另一个表达的NUDX 1基因的特性，从玫瑰品种Rosaxwichurana，RwNUDX 1 -2。为了研究RwNUDX 1 -2蛋白的功能，我们分析了通过杂交R.中国人“老脸红”与R。xwichurana。RwNUDX 1 - 2基因的表达与挥发组的相关性分析表明，RwNUDX 1 - 2基因除了参与其它倍半萜类化合物的生物合成外，还参与一组倍半萜类化合物的生物合成，尤其是E，E-法尼醇的合成。利用子代E，E-法尼醇含量的QTL分析，构建了基于基因标记的遗传图谱。RwNUDX 1 - 2基因与E，E-法尼醇含量QTL共定位，证实了其在R. xwichurana。最后，为了理解玫瑰NUDX蛋白的底物特异性的结构基础，将RhNUDX 1蛋白结晶，并将其结构细化至1.7埃。通过对不同玫瑰NUDX 1蛋白复合物与其各自底物的分子模拟，提出了玫瑰NUDX 1蛋白底物区分的结构基础。
Roses use a non-canonical pathway involving a Nudix hydrolase, RhNUDX1, to synthesize their monoterpenes, especially geraniol. Here we report the characterization of another expressedNUDX1gene from the rose cultivarRosaxwichurana,RwNUDX1-2. In order to study the function of the RwNUDX1-2 protein, we analyzed the volatile profiles of an F(1)progeny generated by crossingR. chinensiscv. 'Old Blush' withR. xwichurana. A correlation test of the volatilomes with gene expression data revealed thatRwNUDX1-2is involved in the biosynthesis of a group of sesquiterpenoids, especiallyE,E-farnesol, in addition to other sesquiterpenes.In vitroenzyme assays and heterologousin plantafunctional characterization of theRwNUDX1-2gene corroborated this result. A quantitative trait locus (QTL) analysis was performed using the data ofE,E-farnesol contents in the progeny and a genetic map was constructed based on gene markers. TheRwNUDX1-2gene co-localized with the QTL forE,E-farnesol content, thereby confirming its function in sesquiterpenoid biosynthesis inR. xwichurana. Finally, in order to understand the structural bases for the substrate specificity of rose NUDX proteins, the RhNUDX1 protein was crystallized, and its structure was refined to 1.7 angstrom. By molecular modeling of different rose NUDX1 protein complexes with their respective substrates, a structural basis for substrate discrimination by rose NUDX1 proteins is proposed.