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Journal of Integrative Agriculture
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Cloning and functional characterization of 1-deoxy-d-xylulose 5-phosphate reductoisomerase (LiDXR) gene in oriental lily (Lilium ‘Sorbonne’)

Xinyue Zhang, Jingqi Dai, Fan Jiang, Tao Yang, Jinzhu Zhang, Jie Dong, Jinping Fan#

College of Horticulture & Landscape Architecture, Northeast Agricultural University, Harbin 150030, China

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摘要  

花香是园林植物重要的观赏形态。萜类化合物中的单萜类是百合花香的主要成分。1-脱氧-D-木酮糖-5-磷酸还原异构酶(DXR)参与了甲基赤藓糖醇磷酸(MEP)途径的第二酶促反应,负责单萜的合成。然而,DXR基因在索邦百合花单萜合成途径中的作用尚不清楚。本研究以东方百合‘索邦’为实验材料,利用早期转录组学数据选择差异表达的LiDXR基因。结果表明,这与百合花期花物质从合成到终止的速率过程具有较高的一致性。因此,我们克隆了LiDXR基因并进行了生物信息学分析,共编码472个氨基酸。LiDXR基因在索邦半开放期表达量最高,花瓣中LiDXR基因的表达量显著高于其他花器官。亚细胞定位结果表明,LiDXR蛋白定位于叶片表皮细胞叶绿体中。病毒诱导的基因沉默(VIGS)实验表明,沉默LiDXR可以通过下调MEP通路下游TPS基因的表达来降低单萜烯水平。同时,HS-SPME-GC-MS分析结果显示,沉默处理后百合花总挥发性萜含量显著降低。过表达植株拟南芥和矮牵牛的结果表明,转基因植株生长潜力更强,开花时间提前。GC-MS结果表明,转基因牵牛花的挥发性总萜含量比野生型高78%。LiDXR基因的过表达会影响MEP通路基因的表达水平,进而影响MEP通路下游包括单萜在内的萜类物质的合成。本研究旨在分析LiDXR基因的功能,为百合及观赏植物的花香育种提供理论依据。



Abstract  

Floral scent is an important ornamental shape of garden plants. Monoterpenes in terpenoids are the main components of lily floral scents. 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) plays a role in the second enzymatic reaction of methylerythritol phosphate (MEP) pathway, which is responsible for monoterpene synthesis. However, the function of DXR gene in the floral monoterpene synthesis pathway of Lilium 'Sorbonne' remains unclear. In this study, the Lilium oriental 'Sorbonne' was used as the experimental material, and the differentially expressed LiDXR gene was selected using the early transcriptomic data. It was found that it had a high consistency with the rater rate process from synthesis to termination of floral substances in the flowering stage of lily. Therefore, the LiDXR gene was cloned and bioinformatics analyzed. A total of 472 amino acids are encoded. The expression of LiDXR gene was the highest at the Sorbonne half opening stage, and the expression of LiDXR gene in petals was significantly higher than that in other flower organs. The results of subcellular localization showed that LiDXR protein was localized in chloroplasts of leaf epidermal cells. A virus-induced gene silencing (VIGS) assay showed that silencing LiDXR can reduce monoterpene levels by down-regulating TPS gene expression downstream of the MEP pathway. Meanwhile, the results of HS-SPME-GC-MS showed that the total volatile terpene content of lily decreased significantly after silenced. The results of overexpressed plants A. thaliana and petunia showed that the transgenic plants had stronger growth potential and advanced flowering time. The GC-MS results of transgenic petunias showed that the content of volatile total terpenes in transgenic strains was 78% higher than that of wild type. Overexpression of LiDXR gene would affect the expression level of MEP pathway genes, and then affect the synthesis of terpenes including monoterpenes downstream of MEP pathway. The purpose of this study was to analyze the function of LiDXR gene and provide theoretical basis for floral breeding of lily and ornamental plants.

Keywords:  Lilium       Floral       Monoterpene       LiDXR  
Online: 07 November 2025  
Fund: 

This research was supported by the Key Research and Development of the Ministry of Science and Technology (2016YFC0500306-02) and the project of the Science and Technology Department of Heilongjiang Province (C2017030).

About author:  Xinyue Zhang, E-mail: 13734523745@163.com; #Correspondence Jinping Fan, Tel: +86-13936646286, E-mail: jinpingfan@neau.edu.cn

Cite this article: 

Xinyue Zhang, Jingqi Dai, Fan Jiang, Tao Yang, Jinzhu Zhang, Jie Dong, Jinping Fan. 2025. Cloning and functional characterization of 1-deoxy-d-xylulose 5-phosphate reductoisomerase (LiDXR) gene in oriental lily (Lilium ‘Sorbonne’). Journal of Integrative Agriculture, Doi:10.1016/j.jia.2025.11.002

Carretero-Paulet L, Ahumada I, Cunillera N, Rodriguez-Concepcion M, Ferrer A, Boronat A, Campos N. 2002. Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiology, 129, 1581-1591.

Carretero-Paulet L, Cairo A, Botella-Pavia P, Besumbes O, Campos N, Boronat A, Rodriguez-Concepcion M. 2006. Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Molecular Biology, 62, 683-695.

Guo X, Li Q, Yan B, Wang Y, Wang S, Xiong F, Zhang C, Zhang Y, Guo L. 2022. Mild shading promotes sesquiterpenoid synthesis and accumulation in Atractylodes lancea by regulating photosynthesis and phytohormones. Scientific reports, 121, 21648.

Hans J, Hause B, Strack D, Walter M H. 2004. Cloning, characterization, and immunolocalization of a mycorrhiza-inducible 1-deoxy-d-xylulose 5-phosphate reductoisomerase in arbuscule-containing cells of maize. Plant Physiology, 134, 614-624.

Jadaun J S, Sangwan N S, Narnoliya L K, Singh N, Bansal S, Mishra B, Sangwan R S. 2017. Over-expression of DXS gene enhances terpenoidal secondary metabolite accumulation in rose-scented geranium and Withania somnifera: active involvement of plastid isoprenogenic pathway in their biosynthesis. Physiologia plantarum, 159, 381-400.

Jiang Y, Qian R, Zhang W, Wei G, Ma X, Zheng J, Kollner T G, Chen F. 2020. Composition and Biosynthesis of Scent Compounds from Sterile Flowers of an Ornamental Plant Clematis florida cv. 'Kaiser'. Molecules, 25, 1711.

Johnson T S, Schwieterman M L, Kim J Y, Cho K H, Clark D G, Colquhoun T A. 2016. Lilium floral fragrance: A biochemical and genetic resource for aroma and flavor. Phytochemistry, 122, 103-112.

Kong Y, Sun M, Pan H, Zhang Q. 2012. Composition and Emission Rhythm of Floral Scent Volatiles from Eight Lily Cut Flowers. Journal of the American Society for Horticultural Science, 137, 376-382.

Kong Y, Wang H, Lang L, Dou X, Bai J. 2022. Effect of Developmental Stages on Genes Involved in Middle and Downstream Pathway of Volatile Terpene Biosynthesis in Rose Petals. Genes, 137, 26-65.

Li Y, Yu C, Mo R, Zhu Z, Dong Z, Hu X, Deng W, Zhuang C. 2022. Screening and Verification of Photosynthesis and Chloroplast-Related Genes in Mulberry by Comparative RNA-Seq and Virus-Induced Gene Silencing. International Journal of Molecular Sciences, 23, 8620.

Mahmoud S S, Croteau R B. 2001. Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylulose phosphate reductoisomerase and menthofuran synthase. Proceedings of the National Academy of Sciences of the United States of America, 98, 8915-8920.

Mallona I, Lischewski S, Weiss J, Hause B, Egea-Cortines M J B P B. 2010. Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida. BMC Plant Biology, 10, 4.

Muhlemann J K, Klempien A, Dudareva N. 2014. Floral volatiles: from biosynthesis to function. Plant, cell & environment, 37, 1936-1949.

Opitz S, Nes W D, Gershenzon J. 2014. Both methylerythritol phosphate and mevalonate pathways contribute to biosynthesis of each of the major isoprenoid classes in young cotton seedlings. Phytochemistry, 98, 110-119.

Orru A, Marchese G, Ruiu S. 2023. Alkaloids in Withania somnifera (L.) Dunal Root Extract Contribute to Its Anti-Inflammatory Activity. Pharmacology, 108, 301-307.

Pichersky E, Noel J P, Dudareva N. 2006. Biosynthesis of plant volatiles: nature's diversity and ingenuity. Science, 311, 808-811.

Pokhilko A, Bou-Torrent J, Pulido P, Rodriguez-Concepcion M, Ebenhoh O. 2015. Mathematical modelling of the diurnal regulation of the MEP pathway in Arabidopsis. The New phytologist, 206, 1075-1085.

Tholl D, Chen F, Petri J, Gershenzon J, Pichersky E. 2005. Two sesquiterpene synthases are responsible for the complex mixture of sesquiterpenes emitted from Arabidopsis flowers. The Plant Journal, 42, 757-771.

Willmer P G, Nuttman C V, Raine N E, Stone G N, Knudsen J T J F E. 2009. Floral volatiles controlling ant behaviour. Functional Ecology, 23, 888-900.

Xing S, Miao J, Li S, Qin G, Tang S, Li H, Gu H, Qu L J. 2010. Disruption of the 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) gene results in albino, dwarf and defects in trichome initiation and stomata closure in Arabidopsis. Cell research, 20, 688-700.

Xu R, Wu J, Zhang Y, Jiang L, Yao J, Zha L, Xie J. 2023. Isolation, characterisation, and expression profiling of DXS and DXR genes in Atractylodes lancea. Genome, 66, 150-164.

Yao H, Gong Y, Zuo K, Ling H, Qiu C, Zhang F, Wang Y, Pi Y, Liu X, Sun X, Tang K. 2008. Molecular cloning, expression profiling and functional analysis of a DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Camptotheca acuminata. Journal of Plant Physiology, 165, 203-213.

Zhang C, Liu H, Hu S, Zong Y, Xia H, Li H. 2022. Transcriptomic profiling of the floral fragrance biosynthesis pathway of Liriodendron and functional characterization of the LtuDXR gene. Plant Science, 314, 111124.

Zhang H, Niu D, Wang J, Zhang S, Yang Y, Jia H, Cui H. 2015. Engineering a Platform for Photosynthetic Pigment, Hormone and Cembrane-Related Diterpenoid Production in Nicotiana tabacum. Plant & cell physiology, 56, 2125-2138.

Zheng B Q, Li X Q, Wang Y. 2023. New Insights into the Mechanism of Spatiotemporal Scent Accumulation in Orchid Flowers. Plants, 12, 304.

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