Promoter Analysis and Transcriptional Profiling of Ginkgo biloba 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase (GbHMGR) gene in Abiotic Stress Responses

LIAO, Yongling; Xu, Feng; Huang, Xiaohua; ZHANG, Weiwei; Cheng, Hua; LI, Linling; Cheng, Shuiyuan; Shen, Yongbao

doi:10.15835/nbha.43.1.9416

Cited by 6 publications

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“…In addition, some genes involved in TTL biosynthesis are upregulated after SA and MeJA treatments are administered in ginkgo. The transcript levels of GbDXS (1-deoxy- d -xylulose 5-phosphate synthase) [ 13 ], GbCMK2 (4-(cytidine 5′-diphospho)-2- C -methyl- d -erythritol kinase) [ 15 ], GbHMGR [ 26 ], GbIDS2 [ 45 ], and GbMVD [ 11 ] are induced by SA and MeJA and are positively involved in TTL biosynthesis in G. biloba . Similar to these reports, our data showed that the expression of GbAACT and GbMVK was upregulated and the TTL content was increased after SA and MeJA treatments, implying that the transcription of these genes responsible for TTL biosynthesis was enhanced.…”

Section: Resultsmentioning

confidence: 99%

“…The expression patterns of GbAACT and GbMVK in various tissues, including roots, stems, leaves, fruits, male and female flowers were also examined to describe TTL synthesis. In addition, our previous work showed that the transcripts of HMGR [ 26 ] and MVD [ 11 ] genes involved in the MVA pathway are positively responsive to methyl jasmonate (MeJA) and salicylic acid (SA) treatments in ginkgo. Therefore, the expression profiles of GbAACT and GbMVK as well as TTL contents under the induction by MeJA and SA were also investigated, which will facilitate future work to map and regulate these important steps involved in TTL biosynthetic pathway at the level of molecular genetics.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba

et al. 2017

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Ginkgolides and bilobalide, collectively termed terpene trilactones (TTLs), are terpenoids that form the main active substance of Ginkgo biloba. Terpenoids in the mevalonate (MVA) biosynthetic pathway include acetyl-CoA C-acetyltransferase (AACT) and mevalonate kinase (MVK) as core enzymes. In this study, two full-length (cDNAs) encoding AACT (GbAACT, GenBank Accession No. KX904942) and MVK (GbMVK, GenBank Accession No. KX904944) were cloned from G. biloba. The deduced GbAACT and GbMVK proteins contain 404 and 396 amino acids with the corresponding open-reading frame (ORF) sizes of 1215 bp and 1194 bp, respectively. Tissue expression pattern analysis revealed that GbAACT was highly expressed in ginkgo fruits and leaves, and GbMVK was highly expressed in leaves and roots. The functional complementation of GbAACT in AACT-deficient Saccharomyces cerevisiae strain Δerg10 and GbMVK in MVK-deficient strain Δerg12 confirmed that GbAACT mediated the conversion of mevalonate acetyl-CoA to acetoacetyl-CoA and GbMVK mediated the conversion of mevalonate to mevalonate phosphate. This observation indicated that GbAACT and GbMVK are functional genes in the cytosolic mevalonate (MVA) biosynthesis pathway. After G. biloba seedlings were treated with methyl jasmonate and salicylic acid, the expression levels of GbAACT and GbMVK increased, and TTL production was enhanced. The cloning, characterization, expression and functional analysis of GbAACT and GbMVK will be helpful to understand more about the role of these two genes involved in TTL biosynthesis.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba

et al. 2017

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“…The presence of G-box and Box 4 elements in the PqHMGR promoter, possibly participating in the light response, has been confirmed. The light-responsive cis -elements have previously been found in promoters of HMGR genes from tomato, maidenhair tree ( Gingko biloba ), mulberry ( Morus alba ), kutki/kutaki ( Picrorhiza kurroa ), red sage ( Salvia miltiorrhiza ), and Andrographis paniculata [36,37,38,39,40,41]. The presence of a G-BOX element suggests that the isolated PqHMGR promoter may be homologous to PgHMGR1 .…”

Section: Discussionmentioning

confidence: 99%

“…Also, the presence of cis -active elements responsive to salicylic acid suggests that the PqHMGR gene may play a role in plant-defense or development-regulation mechanisms [42] (Ma et al 2018). These sequences are rather a common property of the plant HMGR gene regulatory sequences studied in Gingko biloba , Michelia chapensis , Salvia miltiorrhiza , Malus domestica, and tobacco [39,41,43,44,45,46].…”

Section: Discussionmentioning

confidence: 99%

Abscisic Acid Regulates the 3-Hydroxy-3-methylglutaryl CoA Reductase Gene Promoter and Ginsenoside Production in Panax quinquefolium Hairy Root Cultures

Kochan

Balcerczak

Szymczyk

et al. 2019

IJMS

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Panax quinquefolium hairy root cultures synthesize triterpenoid saponins named ginsenosides, that have multidirectional pharmacological activity. The first rate-limiting enzyme in the process of their biosynthesis is 3-hydroxy-3-methylglutaryl CoA reductase (HMGR). In this study, a 741 bp fragment of the P. quinquefolium HMGR gene (PqHMGR), consisting of a proximal promoter, 5′UTR (5′ untranslated region) and 5′CDS (coding DNA sequence) was isolated. In silico analysis of an isolated fragment indicated a lack of tandem repeats, miRNA binding sites, and CpG/CpNpG elements. However, the proximal promoter contained potential cis-elements involved in the response to light, salicylic, and abscisic acid (ABA) that was represented by the motif ABRE (TACGTG). The functional significance of ABA on P. quinquefolium HMGR gene expression was evaluated, carrying out quantitative RT-PCR experiments at different ABA concentrations (0.1, 0.25, 0.5, and 1 mg·L−1). Additionally, the effect of abscisic acid and its time exposure on biomass and ginsenoside level in Panax quinquefolium hairy root was examined. The saponin content was determined using HPLC. The 28 day elicitation period with 1 mg·L−1 ABA was the most efficient for Rg2 and Re (17.38 and 1.83 times increase, respectively) accumulation; however, the protopanaxadiol derivative content decreased in these conditions.

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“…Therefore, the MVA pathway may also have a minimal contribution to the biosynthesis of TTLs. Up to now, some of genes encoding enzymes involved in MVA pathway have been cloned and characterized from G. biloba , including HMGR [ 29 ], mevalonate diphosphate decarboxylase (MVD) [ 10 ], mevalonate kinase (MVK) and acetyl-CoA C -acetyltransferase (AACT) [ 30 ]. However, the HMGS genes in G. biloba are poorly studied.…”

Section: Introductionmentioning

confidence: 99%

Characterization, Function, and Transcriptional Profiling Analysis of 3-Hydroxy-3-methylglutaryl-CoA Synthase Gene (GbHMGS1) towards Stresses and Exogenous Hormone Treatments in Ginkgo biloba

Meng

Song

et al. 2017

Molecules

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3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is one of the rate-limiting enzymes in the mevalonate pathway as it catalyzes the condensation of acetoacetyl-CoA to form 3-hydroxy-3-methylglutaryl-CoA. In this study, A HMGS gene (designated as GbHMGS1) was cloned from Ginkgo biloba for the first time. GbHMGS1 contained a 1422-bp open-reading frame encoding 474 amino acids. Comparative and bioinformatics analysis revealed that GbHMGS1 was extensively homologous to HMGSs from other plant species. Phylogenetic analysis indicated that the GbHMGS1 belonged to the plant HMGS superfamily, sharing a common evolutionary ancestor with other HMGSs, and had a further relationship with other gymnosperm species. The yeast complement assay of GbHMGS1 in HMGS-deficient Saccharomyces cerevisiae strain YSC6274 demonstrated that GbHMGS1 gene encodes a functional HMGS enzyme. The recombinant protein of GbHMGS1 was successfully expressed in E. coli. The in vitro enzyme activity assay showed that the kcat and Km values of GbHMGS1 were 195.4 min−1 and 689 μM, respectively. GbHMGS1 was constitutively expressed in all tested tissues, including the roots, stems, leaves, female flowers, male flowers and fruits. The transcript accumulation for GbHMGS1 was highest in the leaves. Expression profiling analyses revealed that GbHMGS1 expression was induced by abiotic stresses (ultraviolet B and cold) and hormone treatments (salicylic acid, methyl jasmonate, and ethephon) in G. biloba, indicating that GbHMGS1 gene was involved in the response to environmental stresses and plant hormones.

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Promoter Analysis and Transcriptional Profiling of Ginkgo biloba 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase (GbHMGR) gene in Abiotic Stress Responses

Cited by 6 publications

References 0 publications

Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba

Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba

Abscisic Acid Regulates the 3-Hydroxy-3-methylglutaryl CoA Reductase Gene Promoter and Ginsenoside Production in Panax quinquefolium Hairy Root Cultures

Characterization, Function, and Transcriptional Profiling Analysis of 3-Hydroxy-3-methylglutaryl-CoA Synthase Gene (GbHMGS1) towards Stresses and Exogenous Hormone Treatments in Ginkgo biloba

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