SmPPT, a 4-hydroxybenzoate polyprenyl diphosphate transferase gene involved in ubiquinone biosynthesis, confers salt tolerance in Salvia miltiorrhiza
SmPPT, which encodes 4-hydroxybenzoate polyprenyl diphosphate transferase involved in ubiquinone biosynthesis, confers salt tolerance to S. miltiorrhiza through enhancing the activities of POD and CAT to scavenge ROS.
Ubiquinone (UQ), also known as coenzyme Q (CoQ), is a key electron transporter in the mitochondrial respiratory system. UQ is composed of a benzene quinone ring and a polyisoprenoid side chain. Attachment of polyisoprenoid side chain to the benzene quinone ring is a rate-limiting step catalyzed by 4-hydroxybenzoate polyprenyl diphosphate transferase (PPT). So far, only a few plant PPT-encoding genes have been functionally analyzed. Through genome-wide analysis and subsequent molecular cloning, a PPT-encoding gene, termed SmPPT, was identified from an economically and academically important medicinal model plant, Salvia miltiorrhiza. SmPPT contained many putative cis-elements associated with abiotic stresses in the promoter region and were responsive to PEG-6000 and methyl jasmonate treatments. The deduced SmPPT protein contains the PT_UbiA conserved domain of polyprenyl diphosphate transferase and an N-terminal mitochondria transit peptide. Transient expression assay of SmPPT-GFP fusion protein showed that SmPPT was mainly localized in the mitochondria. SmPPT could functionally complement coq2 mutation and catalyzed UQ6 production in yeast cells. Overexpression of SmPPT increased UQ production and enhanced salt tolerance in S. miltiorrhiza. Under salinity stress conditions, transgenic plants accumulated less H2O2 and malondialdehyde and exhibited higher peroxidase (POD) and catalase (CAT) activities compared with wild-type plants. It indicates that SmPPT confers salt tolerance to S. miltiorrhiza at least partially through enhancing the activities of POD and CAT to scavenge ROS.
KeywordsSalvia miltiorrhiza SmPPT Ubiquinone Transgenic plants Salinity stress ROS
This work was supported by the CAMS Innovation Fund for Medical Sciences (CIFMS) (2016-I2M-3-016), the National Natural Science Foundation of China (81773836) and the Fund for Postgraduate Innovation in Peking Union Medical College (2015-1007-14). We appreciate Prof. Xian’en Li for kindly providing S. miltiorrhiza plants and Prof. Wei Xiao for kindly providing yeast coq2 mutant.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Adly AAM (2002) Oxidative stress and disease: an updated review. Res J Immunol 3:129–145Google Scholar
- Araújo WL, Ishizaki K, Nunes-Nesi A, Larson TR, Tohge T, Krahnert I, Witt S, Obata T, Schauer N, Graham IA, Leaver CJ, Fernie AR (2010) Identification of the 2-hydroxyglutarate and isovaleryl-CoA dehydrogenases as alternative electron donors linking lysine catabolism to the electron transport chain of Arabidopsis mitochondria. Plant Cell 22:1549–1563PubMedPubMedCentralGoogle Scholar
- Ashby MN, Kutsunai SY, Ackerman S, Tzagoloff A, Edwards PA (1992) COQ2 is a candidate for the structural gene encoding para-hydroxybenzoate: polyprenyltransferase. J Bio Chem 267:4128–4136Google Scholar
- Büssis D, Heineke D (1998) Acclimation of potato plants to polyethylene glycol-induced water deficit II. Contents and subcellular distribution of organic solutes. J Exp Bot 49:1361–1370Google Scholar
- Chang J, Fu X, An L, Chen T (2006) Properties of cellular ubiquinone and stress-resistance in suspension- cultured cells of Chorispora bungeana during early chilling. Environ Exp Bot 57:116–122Google Scholar
- Dcluzeau A, Wamboldt Y, Elowsky CG, Mackenzie SA, Schuurink RC, Basset GJC (2012) Gene network reconstruction identifies the authentic trans-prenyl diphosphate synthase that makes the solanesyl moiety of ubiquinone-9 in Arabidopsis. Plant J 69:366–375Google Scholar
- Dhindsa RS, Matowe W (1981) Drought tolerance in two mosses: correlated with enzymatic defence against lipid peroxidation. J Exp Bot 32:79–91Google Scholar
- Du Q, Li C, Li D, Lu S (2015) Genome-wide analysis, molecular cloning and expression profiling reveal tissue-specifically expressed, feedback-regulated, stress-responsive and alternatively spliced novel genes involved in gibberellin metabolism in Salvia miltiorrhiza. BMC Genom 16:1087Google Scholar
- Dutta A, Chan SH, Pauli NT, Raina R (2015) HYPERSENSITIVE RESPONSE-LIKE LESIONS 1 Codes for AtPPT1 and regulates accumulation of ROS and defense against bacterial pathogen Pseudomonas syringae in Arabidopsis thaliana. Antioxid Redox Sign 22:785–796Google Scholar
- Ernster L, Dallner G (1995) Biochemical, physiological and medical aspects of ubiquinone function. Biochim Biophy Acta 1271:195–204Google Scholar
- Foyer CH, Noctor G (2003) Redox sensing and signaling associated with reactive oxygen in chloroplast, peroxisomes and mitochondria. Physiol Plant 119:355–364Google Scholar
- Fu XZ, Chen CW, Wang Y, Liu JH, Moriguchi T (2011) Ectopic expression of MdSPDS1 in sweet orange (Citrus sinensis Osbeck) reduces canker susceptibility: involvement of H2O2 production and transcriptional alteration. BMC Plant Bio 11:55Google Scholar
- Hofgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Re 16:9877Google Scholar
- Hou X, Shao F, Ma Y, Lu S (2013) The phenylalanine ammonia-lyase gene family in Salvia miltiorrhiza: genome-wide characterization, molecular cloning and expression analysis. Mol Biol Rep 40:4301–4310Google Scholar
- Lowry OH, Rosebrough NJ, Farr AL (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–276Google Scholar
- Marchler-Bauer A, Derbyshire MK, Gonzales NR, Lu S, Chitsaz F, Geer LY, Geer RC, He J, Gwadz M, Hurwitz DI, Lanczycki CJ, Lu F, Marchler GH, Song JS, Thanki N, Wang Z, Yamashita RA, Zhang D, Zheng C, Bryant SH (2015) CDD: NCBI’s conserved domain database. Nucleic Acids Res 43:D222–D226PubMedGoogle Scholar
- Møller IM (2001) Plant mitochondria and oxidative stress: electron transport, NADPH turnover, and metabolism of reactive oxygen species. Annu Rev Plant Biol 52:561–591Google Scholar
- Moludi J, Keshavarz S, Hosseinzadeh-attar MJ, Frooshani AR, Sadeghpour A, Salarkia S, Gholizadeh F (2015) Coenzyme Q10 effect in prevention of atrial fibrillation after Coronary Artery Bypass Graft: double-blind randomized clinical trial. Tehran Univ Med J 73:79–85Google Scholar
- Okada K, Ohara K, Yazaki K, Nozaki K, Uchida N, Kawamukai M, Nojiri H, Yamane H (2004) The AtPPT1 gene encoding 4-hydroxybenzoate polyprenyl diphosphate transferase in ubiquinone biosynthesis is required for embryo development in Arabidopsis thaliana. Plant Mol Biol 57:567–577Google Scholar
- Song J, Luo H, Li C, Sun C, Xu J, Chen SL (2013) Salvia miltiorrhiza as medicinal model plant. Acta Pharmacol Sin 48:1099–1106Google Scholar
- Wang Y, Hekimi S (2016) Understanding ubiquinone. Trends Cell Biol 5:367–378Google Scholar
- Xu H, Song J, Luo H, Zhang Y, Li Q, Zhu Y, Xu J, Li Y, Song C, Wang B, Sun W, Shen G, Zhang X, Qian J, Ji A, Xu Z, Luo X, He L, Li C, Sun C, Yan H, Cui G, Li X, Li X, Wei J, Liu J, Wang Y, Hayward A, Nelson D, Ning Z, Peter RJ, Qi X, Chen S (2016) Analysis of the genome sequence of the medicinal plant Salvia miltiorrhiza. Mol Plant 9:949–952PubMedPubMedCentralGoogle Scholar
- Yang L, Han H, Liu M, Zuo Z, Zhou K, Lü J, Zhu Y, Bai X, Wang Y (2013) Overexpression of the Arabidopsis photorespiratory pathway gene, serine: glyoxylate aminotransferase (AtAGT1), lead to salt stress tolerance in transgenic duckweed (Lemna minor). Plant Cell Tissue Organ Cult 113:407–416Google Scholar
- Zhang L, Yan X, Wang J, Li S, Liao P, Ka G (2011) Molecular cloning and expression analysis of a new putative gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Salvia miltiorrhiza. Acta Physiol Plant 33:953–961Google Scholar