Tian Lu, Shi Jingwei, Yang Lin, Wei Anzhi
College of Forestry, Northwest A&F University, Yangling, Xianyang 712100, China.
Research Centre for Engineering and Technology of Zanthoxylum State Forestry Administration, Yangling, Xianyang 712100, China.
Foods. 2022 Jun 14;11(12):1746. doi: 10.3390/foods11121746.
Maxim. () has attracted attention for its rich aroma. The aroma of is mainly volatile terpenes synthesized by plant terpene metabolic pathways. However, there is little information on terpene metabolic gene. In this study, the coding sequence of () and () were cloned from cv. 'Fengxiandahongpao.' and genes from with CDS lengths of 2172 bp and 1029 bp, respectively. The bioinformatics results showed that was closely related to citrus, and it was deduced that were hydrophilic proteins without the transmembrane domain. Subcellular localization prediction indicated that was most likely to be located in chloroplasts, and was most likely to be in mitochondria. Meanwhile, the 3D protein structure showed that and were mainly composed of α-helices, which were folded into a single domain. In vitro enzyme activity experiments showed that purified proteins and had the functions of enzyme and enzyme. Transient expression of and in tobacco significantly increased tobacco's terpene content. Moreover, and were expressed in different tissues of , and the relative expression of the two genes was the highest in fruits. Therefore, this suggests that and are positively related to terpene synthesis. This study could provide reference genes for improving breeding as well as for the Rutaceae research.
马克西姆（）因其浓郁的香气而备受关注。的香气主要是植物萜类代谢途径合成的挥发性萜类化合物。然而，关于萜类代谢基因的信息却很少。在本研究中，从‘凤仙大红袍’品种中克隆了（）和（）的编码序列。来自的和基因的编码序列长度分别为2172 bp和1029 bp。生物信息学结果表明，与柑橘密切相关，推测为无跨膜结构域的亲水性蛋白质。亚细胞定位预测表明，最有可能定位于叶绿体中，而最有可能定位于线粒体中。同时，三维蛋白质结构表明，和主要由α-螺旋组成，折叠成单个结构域。体外酶活性实验表明，纯化后的蛋白质和具有酶和酶的功能。和在烟草中的瞬时表达显著提高了烟草的萜类含量。此外，和在的不同组织中表达，且两个基因在果实中的相对表达量最高。因此，这表明和与萜类合成呈正相关。本研究可为改良育种以及芸香科研究提供参考基因。
