Promoter activity analysis and transcriptional profile of Ginkgo biloba 1-Deoxy-D- Xylulose 5-Phosphate reductoisomerase gene (GbDXR) under abiotic stresses

  • Honghui YUAN Nanjing Forestry University, College of Forestry, Nanjing, 210037 (CN)
  • Linling LI Huanggang Normal University, Economic Forest Germplasm Improvement and Comprehensive Utilization of Resources of Hubei Key Laboratories, Huanggang, 438000; Wuhan Polytechnic University, School of Modern Industry for Selenium Science and Engineering, Wuhan 430023 (CN)
  • Li LI Wuhan Polytechnic University, School of Modern Industry for Selenium Science and Engineering, Wuhan 430023 (CN)
  • Hua CHENG Huanggang Normal University, Economic Forest Germplasm Improvement and Comprehensive Utilization of Resources of Hubei Key Laboratories, Huanggang, 438000; Wuhan Polytechnic University, School of Modern Industry for Selenium Science and Engineering, Wuhan 430023 (CN)
  • Shuiyuan CHENG Wuhan Polytechnic University, School of Modern Industry for Selenium Science and Engineering, Wuhan 430023 (CN)
Keywords: DXR promoter, EMSA analysis, Ginkgo biloba, terpene trilactones, transient expression

Abstract

Terpene trilactones (TTL) is a pharmacological ingredient in Ginkgo biloba and its content has become one of the key indices for medicinal value evaluation of ginkgo. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) catalyzes the first step specific for isopentenyl diphosphate production in methylerythritol phosphate pathway, which provide the basic structure required for TTLs biosynthesis. To understand the mechanism controlling the GbDXR gene expression, the GbDXR promoter sequence was isolated and subjected to transient expression with the green fluorescent protein (GFP) in tobacco plants. Characteristic analysis revealed various cis-acting elements that related to light-regulated transcription, hormone signaling (auxin, ethylene), adversity stress and defense signaling (heat/dehydration stress) in the GbDXR promoter region. In transient expression assay, deletion of different portions of the upstream GbDXR promoter identified that the promoter region -3230bp to -865bp conserve the positive regulation function, which could promote the expression of GFP in the cytoplasm of tobacco leaf epidermal cells. The regulation function of the promoter region -865bp to -262bp remained to be elucidated. EMSA analysis suggested possible interactions of GbERF10 and GbERF17 with the ERF-binding elements in the upstream of GbDXR promoter. For abiotic stresses treatment, the expression of GbDXR gene could be significantly induced by UV-B and drought stress. In general, the GbDXR gene expressed differently in different ginkgo tissues but exhibited the highest transcriptional level in the root, with the maximum TTLs content simultaneously. The positive relationship between gene expression level and TTLs content indicated that the GbDXR is responsible for TTLs biosynthesis in G. biloba.

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References

Aird WC, Parvin JD, Sharp PA, Rosenberg RD (1994). The interaction of GATA-binding proteins and basal transcription factors with GATA box-containing core promoters. A model of tissue-specific gene expression. Journal of Biological Chemistry 269:883-889.

Akula R, Ravishankar GA (2011). Influence of abiotic stress signals on secondary metabolites in plants. Plant Signaling & Behavior 6:1720-1731. https://doi.org/10.4161/psb.6.11.17613

Altamura MM (2004). Agrobacterium rhizogenes rolB and rolD genes: regulation and involvement in plant development. Plant Cell, Tissue and Organ Culture 77:89-101. https://doi.org/10.1023/B:TICU.

Bowe LM, Coat G, Pamphilis CW (2000). Phylogeny of seed plants based on all three genomic compartments: extant gymnosperms are monophyletic and Gnetales’ closest relatives are conifers. Proceedings of the National Academy of Sciences of the United States of America 97:4092-4097. https://doi.org/10.1073/pnas.97.8.4092

Burchard P, Bilger W, Weissenböck G (2000). Contribution of hydroxycinnamates and flavonoids to epidermal shielding of UV-A and UV-B radiation in developing rye primary leaves as assessed by ultraviolet-induced chlorophyll fluorescence measurements. Plant, Cell & Environment 23:1373-1380. https://doi.org/10.1046/j.1365-3040.2000.00633.x

Cheng YJ, Guo WW, Yi HL, Pang XM, Deng X (2003). An efficient protocol for genomic DNA extraction from Citrus species. Plant Molecular Biology Reporter 21:177-178. https://doi.org/10.1007/BF02774246

Cordoba E, Salmi M, León P (2009). Unravelling the regulatory mechanisms that modulate the MEP pathway in higher plants. Journal of Experimental Botany 60:2933-2943. https://doi.org/10.1093/jxb/erp190

DeKosky ST, Williamson JD, Fitzpatrick AL, Kronmal RA, Ives DG, Saxton JA, Kuller LH (2008). Ginkgo biloba for prevention of dementia: a randomized controlled trial. Journal of the American Medical Association 300:2253-2262. https://doi.org/10.1001/jama.2008.683

Duan C, Rio M, Leclercq J, Bonnot F, Oliver G, Montoro P (2010). Gene expression pattern in response to wounding, methyl jasmonate and ethylene in the bark of Hevea brasiliensis. Tree Physiology 30:1349-1359. https://doi.org/10.1093/treephys/tpq066

Estévez JM, Cantero A, Reindl A, Reichler S, León P (2001). 1-Deoxy-D-xylulose-5-phosphate synthase, a limiting enzyme for plastidic isoprenoid biosynthesis in plants. The Journal of Biological Chemistry 276:22901-22909. https://doi.org/10.1074/jbc.M100854200

Fu JY, Yang C, Li DW, Tang ZH, Guo XR, Zu YG (2017). Effects of elevated UV-B radiation on photosynthesis and contents of active substance of Eucommia ulmoides plantation. Journal of Anhui Agricultural Sciences 45:6-10. https://doi.org/10.13989/j.cnki.0517-6611.2017. 26.002

Guan LM, Zhao J, Scandalios JG (2000). Cis-elements and transfactors that regulate expression of the maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2O2 is the likely intermediary signaling molecule for the response. The Plant Journal 22:87-95. https://doi.org/10.1046/j.1365-313x.2000.00723.x

Guo YM, Zeng LJ, Liang SY, Deng RQ, Ye HN, Liao SX (2016). Effect of UV-B radiation on growth and main secondary metabolites of Dendrobium officinale Kimura et Migo. Northern Horticulture 17:154-156. https://doi.org/10.11937/bfyy.201617037

Hao GP, Du XH, Shi RJ (2007). Exogenous nitric oxide accelerates soluble sugar, proline and secondary metabolite synthesis in Ginkgo biloba under drought stress. Journal of Plant Physiology and Molecular Biology 33:499-506. https://doi.org/10.1360/aps07042

Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999). Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nature Biotechnology 17:287-291. https://doi.org/10.1038/7036

Kim JH, Lee KI, Chang YJ, Kim SU (2012). Developmental pattern of Ginkgo biloba levopimaradiene synthase (GbLPS) as probed by promoter analysis in Arabidopsis thaliana. Plant Cell Reports 31:1119-1127. https://doi.org/10.1007/s00299-012-1232-1

Kim SM, Kuzuyama T, Chang YJ, Song KS, Kim SU (2006). Identification of class 2 1-deoxy-D-xylulose 5-phosphate synthase and 1-deoxy-D-xylulose 5-phosphate reductoisomerase genes from Ginkgo biloba and their transcription in embryo culture with respect to ginkgolide biosynthesis. Planta Medica 72:234-240. https://doi.org/10.1055/s-2005-916180

Lange BM, Rujan T, Martin W, Croteau R (2000). Isoprenoid biosynthesis: the evolution of two ancient and distinct pathways across genomes. Proceedings of the National Academy of Sciences of the United States of America 97:13172-13177. https://doi.org/10.1073/pnas.240454797

Li W, Fitzloff JF (2002). Simultaneous determination of terpene lactones and flavonoid aglycones in Ginkgo biloba by high-performance liquid chromatography with evaporative light scattering detection. Journal of pharmaceutical and biomedical analysis 30:67-75. https://doi.org/10.1016/S0731-7085(02)00201-7

Liao Y, Xu F, Huang X, Zhang W, Cheng H, Wang X, Shen Y (2016). Characterization and transcriptional profiling of Ginkgo biloba mevalonate diphosphate decarboxylase gene (GbMVD) promoter towards light and exogenous hormone treatments. Plant Molecular Biology Reporter 34:566-581. https://doi.org/10.1007/s11105-015-0947-x

Noguchi M, Miyamoto S, Silverman TA, Safer B (1994). Characterization of an antisense Inr element in the eIF-2 alpha gene. Journal of Biological Chemistry 269:29161-29167. https://doi.org/10.1161/01.CIR.89.3.1320

Perisic O, Lam E (1992). A tobacco DNA binding protein that interacts with a light-responsive box II element. The Plant Cell 4:831-838. https://doi.org/10.1105/tpc.4.7.831

Phillips SM, Dubery IA, Van Heerden H (2013). Molecular characterization of two homoeologous elicitor-responsive lipin genes in cotton. Molecular Genetics and Genomics 288:519-533. https://doi.org/10.1007/s00438-013-0770-8

Pirrello J, Prasad BN, Zhang W, Chen K, Mila I, Zouine M, Bouzayen M (2012). Functional analysis and binding affinity of tomato ethylene response factors provide insight on the molecular bases of plant differential responses to ethylene. BioMed Central Plant Biology 12:190. https://doi.org/10.1186/1471-2229-12-190

Proudhon D, Wei J, Briat JF, Theil EC (1996). Ferritin gene organization: differences between plants and animals suggest possible kingdom-specific selective constraints. Journal of Molecular Evolution 42:325-336. https://doi.org/10.1007/BF02337543

Rodríguez M, Ringstad L, Schäfer P, Just S, Hofer HW, Malmsten M, Siegel G (2007). Reduction of atherosclerotic nanoplaque formation and size by Ginkgo biloba (EGb 761) in cardiovascular high-risk patients. Atherosclerosis 192:438-444. https://doi.org/10.1016/j.atherosclerosis.2007.02.021

Sasaki K, Mitsuhara I, Seo S, Ito H, Matsui H, Ohashi Y (2007). Two novel AP2/ERF domain proteins interact with cis-element VWRE for wound-induced expression of the Tobacco tpoxN1 gene. The Plant Journal 50:1079-1092. https://doi.org/10.1111/j.1365- 313x.2007.03111.x

Sazegari S, Niazi A, Ahmadi FS (2015). A study on the regulatory network with promoter analysis for Arabidopsis DREB-genes. Biomedical Informatics 11:101-106. https://doi.org/10.6026/97320630011101

Si C, Yao XQ, He XL, Chu JZ, Ma CH, Shi XF (2015). Effects of enhanced UV-B radiation on biochemical traits in postharvest flowers of medicinal chrysanthemum. Photochemistry & Photobiology an International Journal 91:845-850. https://doi.org/10.1111/php.12450

Sparkes IA, Runions J, Kearns A, Hawes C (2006). Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nature protocols 1:2019-2025. https://doi.org/10.1038/nprot.2006.286

Strømgaard K, Nakanishi K (2004). Chemistry and biology of terpene trilactones from Ginkgo Biloba. ChemInform 43:1640-1658. https://doi.org/10.1002/anie.200300601

Sun MH, Shi M, Wang Y, Huang Q, Yuan TP, Wang Q, … Kai GY (2019). The biosynthesis of phenolic acids is positively regulated by the JA-responsive transcription factor ERF115 in Salvia miltiorrhiza, Journal of Experimental Botany 70:243-254. https://doi.org/10.1093/jxb/ery349

Takshak S, Agrawal SB (2014). Secondary metabolites and phenylpropanoid pathway enzymes as influenced under supplemental ultraviolet-B radiation in Withania somnifera Dunal, an indigenous medicinal plant. Journal of Photochemistry and Photobiology B: Biology 140:332-343. https://doi.org/10.1016/j.jphotobiol.2014.08.011

Tebbutt SJ, Lonsdale DM (1995). Deletion analysis of a tobacco pollen-specific polygalacturonase promoter. Sexual Plant Reproduction 8:242-246. https://doi.org/10.1007/ BF00228944

Uvackova L, Ondruskova E, Danchenko M, Skultety L, Miernyk J, Hrubík P, Hajduch M (2014). Establishing a leaf proteome reference map for Ginkgo biloba provides insight into potential ethnobotanical uses. Journal of Agricultural and Food Chemistry 62:11547-11556. https://doi.org/10.1021/jf503375a

Vranová E, Coman D, Gruissem W (2012). Structure and dynamics of the isoprenoid pathway network. Molecular Plant 5:318-333. https://doi.org/10.1093/mp/sss015

Watanabe CM, Wolffram S, Ader P, Rimbach G, Packer L, Maguire JJ, Gohil K (2001). The in vivo neuro-modulatory effects of the herbal medicine ginkgo biloba. Proceedings of the National Academy of Sciences 98:6577-6580. https://doi.org/10.1073/pnas.111126298

Yamaguchi-Shinozaki K, Shinozaki K (2005). Organization of cis-acting regulatory elements in osmotic and cold-stress-responsive promoters. Trends in Plant Science 10:88-94. https://doi.org/10.1016/j.tplants.2004.12.012

Yang CQ, Fang X, Wu XM, Mao YB, Wang LJ, Chen XY (2012) Transcriptional regulation of plant secondary metabolism. Journal of Integrative Plant Biology 54:703-712. https://doi.org/10.1111/j.1744-7909.2012.01161.x

Yu ZX, Li JX, Yang CQ, Hu WL, Wang LJ, Chen XY (2012). The jasmonate-responsive AP2/ERF transcription factors AaERF1 and AaERF2 positively regulate artemisinin biosynthesis in Artemisia annua L. Molecular Plant 5:353-365. https://doi.org/10.1093/mp/ssr087

Zhang CJ, Guo IQ, Chen GX, Xie HC (2005). Effects of high temperature and/or drought on growth and secondary metabolites in Ginkgo biloba leaves. Journal of Ecology and Rural Environment 21:11-15. https://doi.org/10.3969/j.issn.1673-4831.2005.03.003

Published
2022-02-24
How to Cite
YUAN, H., LI, L., LI, L., CHENG, H., & CHENG, S. (2022). Promoter activity analysis and transcriptional profile of Ginkgo biloba 1-Deoxy-D- Xylulose 5-Phosphate reductoisomerase gene (GbDXR) under abiotic stresses. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 50(1), 12562. https://doi.org/10.15835/nbha50112562
Section
Research Articles
CITATION
DOI: 10.15835/nbha50112562