Cloning, characterization, and functional analysis of acetyl-CoA C-acetyltransferase and 3-hydroxy-3-methylglutaryl-CoA synthase genes in Santalum album.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
13 01 2021
13 01 2021
Historique:
received:
20
07
2020
accepted:
14
12
2020
entrez:
14
1
2021
pubmed:
15
1
2021
medline:
17
8
2021
Statut:
epublish
Résumé
Sandalwood (Santalum album L.) is famous for its unique fragrance derived from the essential oil of heartwood, whose major components are santalols. To understand the mechanism underlying the biosynthesis of santalols, in this study, we cloned two related genes involved in the mevalonate pathway in S. album coding for acetyl-CoA C-acetyl transferase (AACT) and 3-hydroxy-3-methyglutary-CoA synthase (HMGS). These genes were characterized and functionally analyzed, and their expression profiles were also assessed. An AACT gene designated as SaAACT (GenBank accession No. MH018694) and a HMGS gene designated as SaHMGS (GenBank accession No. MH018695) were successfully cloned from S. album. The deduced SaAACT and SaHMGS proteins contain 415 and 470 amino acids, and the corresponding size of their open-reading frames is 1538 bp and 1807 bp, respectively. Phylogenetic trees showed that the SaAACT protein had the closest relationship with AACT from Hevea brasiliensis and the SaHMGS proteins had the highest homology with HMGS from Siraitia grosvenorii. Functional complementation of SaAACT and SaHMGS in a mutant yeast strain deficient in these proteins confirmed that SaAACT and SaHMGS cDNA encodes functional SaAACT and SaHMGS that mediate mevalonate biosynthesis in yeast. Tissue-specific expression analysis revealed that both genes were constitutively expressed in all examined tissues (roots, sapwood, heartwood, young leaves, mature leaves and shoots) of S. album, both genes showing highest expression in roots. After S. album seedlings were treated with 100 μM methyl jasmonate, the expression levels of SaAACT and SaHMGS genes increased, suggesting that these genes were responsive to this elicitor. These studies provide insight that would allow further analysis of the role of genes related to the sandalwood mevalonate pathway in the regulation of biosynthesis of sandalwood terpenoids and a deeper understanding of the molecular mechanism of santalol biosynthesis.
Identifiants
pubmed: 33441887
doi: 10.1038/s41598-020-80268-3
pii: 10.1038/s41598-020-80268-3
pmc: PMC7807033
doi:
Substances chimiques
Plant Proteins
0
Acetyl-CoA C-Acetyltransferase
EC 2.3.1.9
Hydroxymethylglutaryl-CoA Synthase
EC 2.3.3.10
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1082Références
Harbaugh, D. T. & Baldwin, B. G. Phylogeny and biogeography of the sandalwoods (Santalum, Santalaceae): repeated dispersals throughout the Pacific. Am. J. Bot. 94, 1028–1040 (2007).
pubmed: 21636472
doi: 10.3732/ajb.94.6.1028
Teixeira da Silva, J. A. et al. Sandalwood: basic biology, tissue culture, and genetic transformation. Planta 243, 847–887 (2016).
pubmed: 26745967
doi: 10.1007/s00425-015-2452-8
Benencia, F. & Courrèges, M. C. Antiviral activity of sandalwood oil against Herpes simplex viruses-1 and -2. Phytomed. 6, 119–123 (1999).
doi: 10.1016/S0944-7113(99)80046-4
Solle, H. R. L. & Semiarti, E. Micropropagation of sandalwood (Santalum album L.) endemic plant from East Nusa Tenggara. Indonesia. AIP Conf Proceed 1744, 020026 (2016).
doi: 10.1063/1.4953500
Birkbeck, A. A. The synthesis of fragrant natural products from Santalum album L.: (+)-(Z)-α-santalol and (-)-(Z)-β-santalol. Chimia 71, 823–835 (2017).
pubmed: 29289243
doi: 10.2533/chimia.2017.823
Brand, J. E., Fox, J. E. D., Pronk, G. & Cornwell, C. Comparison of oil concentration and oil quality from Santalum spicatum and S. album plantations, 8–25 years old, with those from mature S. spicatum natural stands. Aust. For. 70, 235–241 (2007).
doi: 10.1080/00049158.2007.10675025
Subasinghe, S. M. C. U. P., Samarasekara, S. C., Millaniyage, K. P. & Hettiarachchi, D. S. Heartwood assessment of natural Santalum album, populations for agroforestry development in Sri Lanka. Agrofor. Syst. 91, 1157–1164 (2017).
doi: 10.1007/s10457-016-0001-5
Jones, C. G., Plummer, J. A. & Barbour, E. L. Non-destructive sampling of Indian sandalwood (Santalum album L.) for oil content and composition. J. Essent. Oil Res. 19, 157–164 (2007).
doi: 10.1080/10412905.2007.9699250
Howes, M. J., Simmonds, M. S. J. & Kite, G. C. Evaluation of the quality of sandalwood essential oils by gas chromatography-mass spectrometry. J. Chromatogr. A 1028, 307–312 (2004).
pubmed: 14989484
doi: 10.1016/j.chroma.2003.11.093
Demole, E., Demole, C. & Enggist, P. A chemical investigation of the volatile constituents of east Indian sandalwood oil (Santalum album L.). Helvet. Chim. Acta 59, 737–747 (2010).
doi: 10.1002/hlca.19760590304
Baldovini, N., Delasalle, C. & Joulain, D. Phytochemistry of the heartwood from fragrant Santalum species: a review. Flav. Frag. J. 26, 7–26 (2011).
doi: 10.1002/ffj.2025
Hasegawa, T. et al. Isolation of new constituents with a formyl group from the heartwood of Santalum album L. Flav. Frag. J. 26, 98–100 (2015).
doi: 10.1002/ffj.2020
Lichtenthaler, H. K. The 1-deoxy-D-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Ann. Rev. Plant Biol. 50, 47–65 (1999).
doi: 10.1146/annurev.arplant.50.1.47
Rohmer, M. The discovery of a mevalonate-independent pathway for isoprenoid biosynthesis in bacteria, algae and higher plants. Nat. Prod. Rep. 16, 565–574 (1999).
pubmed: 10584331
doi: 10.1039/a709175c
Lange, B. M., Rujan, T., Martin, W. & Croteau, R. Isoprenoid biosynthesis: the evolution of two ancient and distinct pathways across genomes. Proc. Nat. Acad. Sci. USA 97, 13172–13177 (2000).
pubmed: 11078528
doi: 10.1073/pnas.240454797
pmcid: 27197
Kuzuyama, T. Mevalonate and nonmevalonate pathways for the biosynthesis of isoprene units. Biosci. Biotech. Biochem. 66, 1619–1627 (2002).
doi: 10.1271/bbb.66.1619
Estévez, J. M., Cantero, A., Reindl, A., Reichler, S. & León, P. 1-Deoxy-D-xylulose-5-phosphate synthase, a limiting enzyme for plastidic isoprenoid biosynthesis in plants. J. Biol. Chem. 276, 22901–22909 (2001).
pubmed: 11264287
doi: 10.1074/jbc.M100854200
Dubey, V. S., Bhalla, R. & Luthra, R. An overview of the non-mevalonate pathway for terpenoid biosynthesis in plants. J. Biosci. 28, 637–646 (2003).
pubmed: 14517367
doi: 10.1007/BF02703339
Gutensohn, M. et al. Cytosolic monoterpene biosynthesis is supported by plastid-generated geranyl diphosphate substrate in transgenic tomato fruits. Plant J. 75, 351–363 (2013).
pubmed: 23607888
doi: 10.1111/tpj.12212
Laule, O. et al. Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. Proc. Nat. Acad. Sci. USA 100, 6866–6871 (2003).
pubmed: 12748386
doi: 10.1073/pnas.1031755100
pmcid: 164538
Miziorko, H. M. Enzymes of the mevalonate pathway of isoprenoid biosynthesis. Arch. Biochem. Biophys. 505, 131–143 (2011).
pubmed: 20932952
pmcid: 3026612
doi: 10.1016/j.abb.2010.09.028
Tholl, D. Biosynthesis and biological functions of terpenoids in plants. Adv. Bioch. Engin. Biotech. 148, 63–106 (2015).
Soto, G. et al. Acetoacetyl-CoA thiolase regulates the mevalonate pathway during abiotic stress adaptation. J. Exp. Bot. 62, 5699–5711 (2011).
pubmed: 21908473
doi: 10.1093/jxb/err287
Luskey, K. L. & Stevens, B. Human 3-hydroxy-3-methylglutaryl coenzyme A reductase. Conserved domains responsible for catalytic activity and sterol-regulated degradation. J. Biol. Chem. 260, 10271–10277 (1985).
pubmed: 2991281
doi: 10.1016/S0021-9258(17)39242-6
Caelles, C., Ferrer, A., Balcells, L., Hegardt, F. G. & Boronat, A. Isolation and structural characterization of a cDNA encoding Arabidopsis thaliana 3-hydroxy-3-methylglutaryl coenzyme-A reductase. Plant Mol. Biol. 13, 627–638 (1990).
doi: 10.1007/BF00016018
Riou, C., Tourte, Y., Lacroute, F. & Karsta, F. Isolation and characterization of a cDNA encoding Arabidopsis thaliana, mevalonate kinase by genetic complementation in yeast. Gene 148, 293–297 (1994).
pubmed: 7958957
doi: 10.1016/0378-1119(94)90701-3
Montamat, F., Guilloton, M., Karst, F. & Delrot, S. Isolation and characterization of a cDNA encoding Arabidopsis thaliana 3-hydroxy-3-methylglutaryl-coenzyme A synthase. Gene 167, 197–201 (1995).
pubmed: 8566777
doi: 10.1016/0378-1119(95)00642-7
Lluch, M. A., Masferrer, A., Arró, M., Boronat, A. & Ferrer, A. Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana. Plant Mol. Biol. 42, 365–376 (2000).
pubmed: 10794536
doi: 10.1023/A:1006325630792
Carrie, C., Murcha, M. W., Millar, A. H., Smith, S. M. & Whelan, J. Nine 3-ketoacyl-CoA thiolases (KATs) and acetoacetyl-CoA thiolases (ACATs) encoded by five genes in Arabidopsis thaliana are targeted either to peroxisomes or cytosol but not to mitochondria. Plant Mol. Biol. 63, 97–108 (2007).
pubmed: 17120136
doi: 10.1007/s11103-006-9075-1
Ahumada, I. et al. Characterisation of the gene family encoding acetoacetyl-CoA thiolase in Arabidopsis. Func. Plant Biol. 35, 1100–1111 (2008).
doi: 10.1071/FP08012
Dai, Z. B., Cui, G. H., Zhou, S. F., Zhang, X. N. & Huang, L. Q. Cloning and characterization of a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Salvia miltiorrhiza involved in diterpenoid tanshinone accumulation. J. Plant Physiol. 168, 148–157 (2011).
pubmed: 20637524
doi: 10.1016/j.jplph.2010.06.008
Xu, Y. H. et al. Identification of genes related to agarwood formation: transcriptome analysis of healthy and wounded tissues of Aquilaria sinensis. BMC Genom. 14, 227 (2013).
doi: 10.1186/1471-2164-14-227
Chye, M. L., Tan, C. T. & Chua, N. H. Three genes encode 3-hydroxy-3-methylglutaryl-coenzyme A reductase in Hevea brasiliensis: hmg1 and hmg3 are differentially expressed. Plant Mol. Biol. 19, 473–484 (1992).
pubmed: 1377968
doi: 10.1007/BF00023395
Sirinupong, N., Suwanmanee, P., Doolittle, R. F. & Suvachitanont, W. Molecular cloning of a new cDNA and expression of 3-hydroxy-3-methylglutaryl-CoA synthase gene from Hevea brasiliensis. Planta 221, 502–512 (2005).
pubmed: 15744497
doi: 10.1007/s00425-004-1463-7
Sando, T. et al. Cloning and characterization of mevalonate pathway genes in a natural rubber producing plant, Hevea brasiliensis. Biosci. Biotechnol. Biochem. 72, 2049–2060 (2008).
pubmed: 18685207
doi: 10.1271/bbb.80165
Shen, G. A. et al. Cloning and characterization of a root-specific expressing gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from Ginkgo biloba. Mol. Biol. Rep. 33, 117–127 (2006).
pubmed: 16817021
doi: 10.1007/s11033-006-0014-7
Tao, T. T. et al. Molecular cloning, characterization, and functional analysis of a gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A synthase from Matricaria chamomilla. Genes Genom. 38, 1179–1187 (2016).
doi: 10.1007/s13258-016-0463-x
Chen, Q. W. et al. Molecular cloning, characterization, and functional analysis of acetyl-CoA C-acetyltransferase and mevalonate kinase genes involved in terpene trilactone biosynthesis from Ginkgo biloba. Molecules 22, 74 (2017).
pmcid: 6155782
doi: 10.3390/molecules22010074
Jones, C. G. et al. Isolation of cDNAs and functional characterisation of two multi-product terpene synthase enzymes from sandalwood, Santalum album L. Arch. Biochem. Biophys. 477, 121–130 (2008).
pubmed: 18541135
doi: 10.1016/j.abb.2008.05.008
Jones, C. G. et al. Sandalwood fragrance biosynthesis involves sesquiterpene synthases of both the terpene synthase (TPS)-a and TPS-b subfamilies, including santalene synthases. J. Biol. Chem. 286, 17445–17454 (2011).
pubmed: 21454632
pmcid: 3093818
doi: 10.1074/jbc.M111.231787
Rani, A., Ravikumar, P., Reddy, M. D. & Kush, A. Molecular regulation of santalol biosynthesis in Santalum album L. Gene 527, 642–648 (2013).
pubmed: 23860319
doi: 10.1016/j.gene.2013.06.080
Srivastava, P. L. et al. Functional characterization of novel sesquiterpene synthases from Indian sandalwood, Santalum album. Sci. Rep. 5, 10095 (2015).
pubmed: 25976282
pmcid: 4432371
doi: 10.1038/srep10095
Diaz-Chavez, M. L. et al. Biosynthesis of sandalwood oil: Santalum album CYP76F cytochromes P450 produce santalols and bergamotol. PLoS ONE 8, e75053 (2013).
pubmed: 24324844
pmcid: 3854609
doi: 10.1371/journal.pone.0075053
Celedon, J. M. et al. Heartwood-specific transcriptome and metabolite signatures of tropical identity sandalwood (Santalum album) reveal the final step of (Z)-santalol fragrance biosynthesis. Plant J. 86, 289–299 (2016).
pubmed: 26991058
doi: 10.1111/tpj.13162
Kai, G. et al. Molecular cloning and expression analyses of a new gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Taxus× media. Biol. Plant. 50, 359–366 (2006).
doi: 10.1007/s10535-006-0050-0
Anderson, V. E., Bahnson, B. J., Wlassics, I. D. & Walsh, C. T. The reaction of acetyldithio-CoA, a readily enolized analog of acetyl-CoA with thiolase from Zoogloea ramigera. J. Biol. Chem. 265, 6255–6261 (1990).
pubmed: 2180945
doi: 10.1016/S0021-9258(19)39318-4
Price, A. C. et al. Inhibition of β-ketoacyl-acyl carrier protein synthases by thiolactomycin and cerulenin structure and mechanism. J. Biol. Chem. 276, 6551–6559 (2001).
pubmed: 11050088
doi: 10.1074/jbc.M007101200
Theisen, M. J. et al. 3-hydroxy- 3-methylglutaryl-CoA synthase intermediate complex observed in “real-time”. Proc. Nat. Acade. Sci. USA 101, 16442–16447 (2004).
doi: 10.1073/pnas.0405809101
Campobasso, N. et al. Staphylococcus aureus 3-hydroxy-3-methylglutaryl-CoA synthase. J. Biol. Chem. 279, 44883–44888 (2004).
pubmed: 15292254
doi: 10.1074/jbc.M407882200
Ahumada, I. et al. Characterization of the gene family encoding acetoacetyl-CoA thiolase in Arabidopsis. Func. Plant Biol. 35, 1100–1111 (2008).
doi: 10.1071/FP08012
Nagegowda, D. A. Plant volatile terpenoid metabolism: Biosynthetic genes, transcriptional regulation and subcellular compartmentation. FEBS Lett. 584, 2965–2973 (2010).
pubmed: 20553718
doi: 10.1016/j.febslet.2010.05.045
Nagegowda, D. A., Ramalingam, S., Hemmerlin, A., Bach, T. J. & Chye, M. L. Brassica juncea HMG-CoA synthase: localization of mRNA and protein. Planta 221, 844–856 (2005).
pubmed: 15770484
doi: 10.1007/s00425-005-1497-5
Trocha, P. J. & Sprinson, D. B. Location and regulation of early enzymes of sterol biosynthesis in yeast. Arch. Biochem. Bioph. 174, 45–51 (1976).
doi: 10.1016/0003-9861(76)90322-2
Servouse, M. & Karst, F. Regulation of early enzymes of ergosterol biosynthesis in Saccharomyces cerevisiae. Biochem. J. 240, 541–547 (1986).
pubmed: 2880580
pmcid: 1147448
doi: 10.1042/bj2400541
Vishwakarma, R. K. et al. Molecular cloning, biochemical characterization, and differential expression of an acetyl-CoA C-acetyltransferase gene (AACT) of brahmi (Bacopa monniera). Plant Mol. Biol. Rep. 31, 547–557 (2013).
doi: 10.1007/s11105-012-0523-6
Pauwels, L. et al. Mapping methyl jasmonate-mediated transcriptional reprogramming of metabolism and cell cycle progression in cultured Arabidopsis cells. Proc. Nati. Acad. Sci. USA 105, 1380–1385 (2008).
doi: 10.1073/pnas.0711203105
Robert-Seilaniantz, A., Grant, M. & Jones, J. D. G. Hormone crosstalk in plant disease and defense: more than just jasmonate-salicylate antagonism. Ann. Rev. Phytopath. 49, 317–343 (2011).
doi: 10.1146/annurev-phyto-073009-114447
Zhang, L. et al. Molecular cloning and expression analysis of a new putative gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Salvia miltiorrhiza. Acta Physiol. Plant. 33, 953–961 (2011).
doi: 10.1007/s11738-010-0627-2
Liu, Y. J. et al. Cloning and characterisation of the gene encoding, 3-hydroxy-3-methylglutaryl-CoA synthase in Tripterygium wilfordii. Molecules 19, 19696–19707 (2014).
pubmed: 25438080
pmcid: 6271793
doi: 10.3390/molecules191219696
Brocke, C., Eh, M. & Finke, A. Recent developments in the chemistry of sandalwood odorants. Chem. Biodiv. 5, 1000–1010 (2008).
doi: 10.1002/cbdv.200890080
Muratore, A., Clinet, J. C. & Dunach, E. Synthesis of new exo- and endo-3,8-dihydro-β-santalols and other norbornyl-derived alcohols. Chem. Biodiv. 7, 623–638 (2010).
doi: 10.1002/cbdv.200900119
Zhang, Y. Y. et al. Multiple strategies for increasing yields of essential oil and obtaining sandalwood terpenoids by biotechnological methods in sandalwood. Trees 32, 21–29 (2018).
doi: 10.1007/s00468-017-1558-y
Oldfield, S., Lusty, C., MacKinven, A. The World List of Threatened Trees, WCMC, IUCN, 632 (1998).
Dooley, K. A., Millinder, S. & Osborne, T. F. Sterol regulation of 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene through a direct interaction between sterol regulatory element binding protein and the trimeric CCAAT-binding factor/nuclear factor Y. J. Biol. Chem. 273, 1349–1356 (1998).
pubmed: 9430668
doi: 10.1074/jbc.273.3.1349
Fang, X. et al. The cloning, characterization and functional analysis of a gene encoding an acetyl-CoA acetyltransferase involved in triterpene biosynthesis in Ganoderma lucidum. Mycoence 54, 100–105 (2013).
Pütter, K. M., Deenen, N. V., Unland, K., Prüfer, D. & Gronover, C. S. Isoprenoid biosynthesis in dandelion latex is enhanced by the overexpression of three key enzymes involved in the mevalonate pathway. BMC Plant Biol. 17, 88 (2017).
pubmed: 28532507
pmcid: 5441070
doi: 10.1186/s12870-017-1036-0
Peralta-Yahya, P. P. et al. Identification and microbial production of a terpene-based advanced biofuel. Nat. Comm. 2, 483 (2011).
doi: 10.1038/ncomms1494
Wang, H., Nagegowda, A. D., Rawat, R., Bouvier-Navé, P., Guo, D. J., Bach, T. J. & Chye, M. L. Overexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance. Plant Biotech. J. 31–42 (2012).
Burdock, G. A. & Carabin, I. G. Safety assessment of sandalwood oil (Santalum album L.). Food Chem. Toxicol. 46, 421–432 (2008).
pubmed: 17980948
doi: 10.1016/j.fct.2007.09.092
Zhang, X. H., Teixeira da Silva, J. A., Jia, Y. X., Yan, J. & Ma, G. H. Essential oil composition from roots of Santalum album L.. J Essent Oil Bear. Plants 15, 1–6 (2012).
doi: 10.1080/0972060X.2012.10644011
Zhang, X. H. et al. Physiological and transcriptomic analyses reveal a response mechanism to cold stress in Santalum album L. leaves. Sci. Rep. 7, 42165 (2017).
pubmed: 28169358
pmcid: 5294638
doi: 10.1038/srep42165
Saitou, N. & Nei, M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406–425 (1987).
pubmed: 3447015
Kumar, S., Stecher, G. & Tamura, K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 33, 1870–1874 (2016).
pubmed: 27004904
doi: 10.1093/molbev/msw054
pmcid: 8210823
Yoo, S. D., Cho, Y. H. & Sheen, J. Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat. Protoc. 2, 1565–1572 (2007).
pubmed: 17585298
doi: 10.1038/nprot.2007.199
Hiser, L., Basson, M. E. & Rine, J. ERG10 from Saccharomyces cerevisiae encodes acetoacetyl-CoA thiolase. J. Biol. Chem. 269, 31383–31389 (1994).
pubmed: 7989303
doi: 10.1016/S0021-9258(18)31705-8
Albers, E. & Larsson, C. A comparison of stress tolerance in YPD and industrial lignocellulose-based medium among industrial and laboratory yeast strains. J. Indus. Microbiol. Biotech. 36, 1085–1091 (2009).
doi: 10.1007/s10295-009-0592-1
Zhang, X. H. et al. RNA-Seq analysis identifies key genes associated with haustorial development in the root hemiparasite Santalum album. Front. Plant Sci. 6, 661 (2015).
pubmed: 26388878
pmcid: 4555033
doi: 10.3389/fpls.2015.00661
Schmittgen, T. D. & Livak, K. J. Analyzing real-time PCR data by the comparative CT method. Nat. Protoc. 3, 1101–1108 (2008).
pubmed: 18546601
doi: 10.1038/nprot.2008.73
Zhang, X. H. et al. Identification and functional characterization of three new terpene synthase genes involved in chemical defense and abiotic stresses in Santalum album. BMC Plant Biol. 19, 115 (2019).
pubmed: 30922222
pmcid: 6437863
doi: 10.1186/s12870-019-1720-3