Methyl jasmonate-induced accumulation of metabolites and transcriptional responses involved in triterpene biosynthesis in Siraitia grosvenorii fruit at different growing stages

Kailun Zhang, Zuliang Luo, Yuhua Guo, Changming Mo, Dongping Tu, Xiaojun Ma, Longhua Bai


The cucurbitane-type triterpenoid glycosides, mogrosides, are the main active components of Siraitia grosvenorii fruit. Squalene and cucurbitadienol are among the intermediates of the biosynthetic pathway for the formation of cucurbitane-type triterpenoid backbones of mogrosides. It is recognized that the exogenous application of methyl jasmonate (MeJA) increases the accumulation of secondary metabolites in various plant species. Here, the effect of MeJA (50, 200, and 500 μM) on the accumulation of squalene and cucurbitadienol in the fruits of S. grosvenorii at 10, 20, and 30 days after flowering (DAF) was tested for the first time. Since mogroside II E is the main cucurbitane-type triterpenoid present at this time, its concentration was also determined. The results show that MeJA can indeed promote squalene and cucurbitadienol accumulation, the application of 500 μM MeJA at 30 DAF being optimal. The concentration of squalene and cucurbitadienol increased up to 0.43 and 4.71 μg/g dry weight (DW), respectively, both of which were 1.2-fold greater than that of the control. The content of mogroside II E increased by 15% over the untreated group. We subsequently analyzed the expression of key genes involved in the mogroside biosynthetic pathway, including the 3-hydroxy-3-methylglutaryl-coenzyme A reductase gene (SgHMGR), squalene synthetase gene (SgSQS), cucurbitadienol synthase gene (SgCS), and cytochrome P450 (SgCYP450) with quantitative real-time PCR. The results showed that transcriptional levels of these genes were upregulated following the treatment described above. Additionally, their responses in the presence of MeJA was related to the concentration and timing of MeJA treatment.


Siraitia grosvenorii; methyl jasmonate (MeJA); squalene; cucurbitadienol; mogroside II E

Full Text:



Philippe RN, de Mey M, Anderson J, Ajikumar PK. Biotechnological production of natural zero-calorie sweeteners. Curr Opin Biotechnol. 2014;26:155–161.

Tang Q, Ma X, Mo C, Wilson IW, Song C, Zhao H, et al. An efficient approach to finding Siraitia grosvenorii triterpene biosynthetic genes by RNA-seq and digital gene expression analysis. BMC Genomics. 2011;12:343.

Deng F, Liang X, Yang L, Liu Q, Liu H. Analysis of mogroside V in Siraitia grosvenorii with micelle-mediated cloud-point extraction. Phytochem Anal. 2013;24(4):381–385.

Li C, Lin LM, Sui F, Wang ZM, Huo HR, Dai L, et al. Chemistry and pharmacology of Siraitia grosvenorii: a review. Chin J Nat Med. 2014;12(2):89–102.

Liu DD, Ji XW, Li RW. Effects of Siraitia grosvenorii fruits extracts on physical fatigue in mice. Iran J Pharm Res. 2013;12(1):115–121.

Pawar RS, Krynitsky AJ, Rader JI. Sweeteners from plants – with emphasis on Stevia rebaudiana (Bertoni) and Siraitia grosvenorii (Swingle). Anal Bioanal Chem. 2013;405(13):4397–4407.

Vranova E, Coman D, Gruissem W. Network analysis of the MVA and MEP pathways for isoprenoid synthesis. Annu Rev Plant Biol. 2013;64:665–700.

Wang L, Yang Z, Lu F, Liu J, Song Y, Li D. Cucurbitane glycosides derived from mogroside IIE: structure–taste relationships, antioxidant activity, and acute toxicity. Molecules. 2014;19(8):12676–12689.

Cárdeno A, Aparicio-Soto M, Montserrat-de la Paz S, Bermudez B, Muriana FJG, Alarcón-de-la-Lastra C. Squalene targets pro- and anti-inflammatory mediators and pathways to modulate over-activation of neutrophils, monocytes and macrophages. J Funct Foods. 2015;14:779–790.

Chen Q, Cheng Z, Yang J, Yi X. The HPLC Analysis of the squalene in Siraitia grosvenorii seed oil. Guangxi Sciences. 2006;13(2):118–120.

Kohno Y, Egawa Y, Itoh S, Nagaoka S, Takahashi M, Mukai K. Kinetic study of quenching reaction of singlet oxygen and scavenging reaction of free radical by squalene in n-butanol. Biochim Biophys Acta. 1995;1256(1):52–56.

Miettinen TA, Vanhanen H. Serum concentration and metabolism of cholesterol during rapeseed oil and squalene feeding. Am J Clin Nutr. 1994;59(2):356–363.

Kelly GS. Squalene and its potential clinical uses. Altern Med Rev. 1999;4(1):29–36.

Rude MA, Schirmer A. New microbial fuels: a biotech perspective. Curr Opin Microbiol. 2009;12(3):274–281.

Hoang MH, Ha NC, Thom le T, Tam LT, Anh HT, Thu NT, et al. Extraction of squalene as value-added product from the residual biomass of Schizochytrium mangrovei PQ6 during biodiesel producing process. J Biosci Bioeng. 2014;118(6):632–639.

Kim SR, Seo HS, Choi HS, Cho SG, Kim YK, Hong EH, et al. Trichosanthes kirilowii ethanol extract and Cucurbitacin D inhibit cell growth and induce apoptosis through inhibition of STAT3 activity in breast cancer cells. Evid Based Complement Alternat Med. 2013;2013:975350.

Shang Y, Ma Y, Zhou Y, Zhang H, Duan L, Chen H, et al. Biosynthesis, regulation, and domestication of bitterness in cucumber. Science. 2014;346(6213):1084–1088.

Theis N, Kesler K, Adler LS. Leaf herbivory increases floral fragrance in male but not female Cucurbita pepo subsp. texana (Cucurbitaceae) flowers. Am J Bot. 2009;96(5):897–903.

Ukiya M, Akihisa T, Tokuda H, Toriumi M, Mukainaka T, Banno N, et al. Inhibitory effects of cucurbitane glycosides and other triterpenoids from the fruit of Momordica grosvenori on Epstein–Barr virus early antigen induced by tumor promoter 12-O-tetradecanoylphorbol-13-acetate. J Agric Food Chem. 2002;50(23):6710–6715.

Akihisa T, Yasukawa K, Kimura Y, Takido M, Kokke WCMC, Tamura T. 7-Oxo-10α-cucurbitadienol from the seeds of Trichosanthes kirilowii and its anti-inflammatory effect. Phytochemistry. 1994;36(1):153–157.

Pauwels L, Inze D, Goossens A. Jasmonate-inducible gene: what does it mean? Trends Plant Sci. 2009;14(2):87–91.

Cheong J-J, Choi YD. Methyl jasmonate as a vital substance in plants. Trends Genet. 2003;19(7):409–413.

Pauwels L, Morreel K, de Witte E, Lammertyn F, van Montagu M, Boerjan W, et al. Mapping methyl jasmonate-mediated transcriptional reprogramming of metabolism and cell cycle progression in cultured Arabidopsis cells. Proc Natl Acad Sci USA. 2008;105(4):1380–1385.

Rischer H, Oresic M, Seppanen-Laakso T, Katajamaa M, Lammertyn F, Ardiles-Diaz W, et al. Gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells. Proc Natl Acad Sci USA. 2006;103(14):5614–5619.

Concha CM, Figueroa NE, Poblete LA, Onate FA, Schwab W, Figueroa CR. Methyl jasmonate treatment induces changes in fruit ripening by modifying the expression of several ripening genes in Fragaria chiloensis fruit. Plant Physiol Biochem. 2013;70:433–444.

Khokon MA, Salam MA, Jammes F, Ye W, Hossain MA, Uraji M, et al. Two guard cell mitogen-activated protein kinases, MPK9 and MPK12, function in methyl jasmonate-induced stomatal closure in Arabidopsis thaliana. Plant Biol (Stuttg) 2015;17(5):946–952.

Zhao J, Davis LC, Verpoorte R. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv. 2005;23(4):283–333.

Bonfill M, Mangas S, Moyano E, Cusido RM, Palazón J. Production of centellosides and phytosterols in cell suspension cultures of Centella asiatica. Plant Cell Tissue Organ Cult. 2010;104(1):61–67.

Hayashi H, Huang P, Inoue K. Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra. Plant Cell Physiol. 2003;44(4):404–411.

Dewey RE, Xie J. Molecular genetics of alkaloid biosynthesis in Nicotiana tabacum. Phytochemistry. 2013;94:10–27.

Fang X, Shi L, Ren A, Jiang AL, Wu FL, Zhao MW. The cloning, characterization and functional analysis of a gene encoding an acetyl-CoA acetyltransferase involved in triterpene biosynthesis in Ganoderma lucidum. Mycoscience. 2013;54(2):100–105.

Han JY, In JG, Kwon YS, Choi YE. Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng. Phytochemistry. 2010;71(1):36–46.

Niu Y, Luo H, Sun C, Yang TJ, Dong L, Huang L, et al. Expression profiling of the triterpene saponin biosynthesis genes FPS, SS, SE, and DS in the medicinal plant Panax notoginseng. Gene. 2014;533(1):295–303.

Chappell J. The biochemistry and molecular biology of isoprenoid metabolism. Plant Physiol. 1995;107(1):1–6.

Spanova M, Daum G. Squalene – biochemistry, molecular biology, process biotechnology, and applications. Eur J Lipid Sci Technol. 2011;113(11):1299–1320.

Shibuya M, Adachi S, Ebizuka Y. Cucurbitadienol synthase, the first committed enzyme for cucurbitacin biosynthesis, is a distinct enzyme from cycloartenol synthase for phytosterol biosynthesis. Tetrahedron. 2004;60(33):6995–7003.

Guo Q, Ma X, Bai L, Pan L, Feng S, Mo C. Screening of reference genes in Siraitia grosvenorii and spatio-temporal expression analysis of 3-hydroxy-3-methylglutaryl coenzyme A. Zhong Cao Yao. 2014;45(15):2224–2229.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2[−Delta Delta C(T)] method. Methods. 2001;25(4):402–408.

Liu JL, Dian Peng LI, Huang YL, Hai Xiao LU. Determination of mogrol glycosides from fruits of Siraitia grosvenorii in different growing ages by HPLC. Guihaia. 2007;27(4):665–668.

Choi DW, Jung JD, Ha YI, Park HW, Dong SI, Chung HJ, et al. Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites. Plant Cell Rep. 2005;23(8):557–566.

Kim OT, Bang KH, Jung SJ, Kim YC, Hyun DY, Kim SH, et al. Molecular characterization of ginseng farnesyl diphosphate synthase gene and its up-regulation by methyl jasmonate. Biol Plant. 2010;54(1):47–53.

Huang X, Li J, Shang H, Meng X. Effect of methyl jasmonate on the anthocyanin content and antioxidant activity of blueberries during cold storage. J Sci Food Agric. 2015;95(2):337–343.

Saavedra GM, Figueroa NE, Poblete LA, Cherian S, Figueroa CR. Effects of preharvest applications of methyl jasmonate and chitosan on postharvest decay, quality and chemical attributes of Fragaria chiloensis fruit. Food Chem. 2016;190:448–453.

Sivankalyani V, Feygenberg O, Maorer D, Zaaroor M, Fallik E, Alkan N. Combined treatments reduce chilling injury and maintain fruit quality in avocado fruit during cold quarantine. PloS One. 2015;10(10):e0140522.

Briceno Z, Almagro L, Sabater-Jara AB, Calderon AA, Pedreno MA, Ferrer MA. Enhancement of phytosterols, taraxasterol and induction of extracellular pathogenesis-related proteins in cell cultures of Solanum lycopersicum cv. Micro-Tom elicited with cyclodextrins and methyl jasmonate. J Plant Physiol. 2012;169(11):1050–1058.

Sabater-Jara AB, Almagro L, Belchi-Navarro S, Ferrer MA, Barcelo AR, Pedreno MA. Induction of sesquiterpenes, phytoesterols and extracellular pathogenesis-related proteins in elicited cell cultures of Capsicum annuum. J Plant Physiol. 2010;167(15):1273–1281.

Suh HW, Hyun SH, Kim SH, Lee SY, Choi HK. Metabolic profiling and enhanced production of phytosterols by elicitation with methyl jasmonate and silver nitrate in whole plant cultures of Lemna paucicostata. Process Biochem. 2013;48(10):1581–1586.

Ciddi V, Srinivasan V, Shuler ML. Elicitation of Taxus sp. cell cultures for production of taxol. Biotechnol Lett. 1995;17(12):1343–1346.

Lu M, Wong H, Teng W. Effects of elicitation on the production of saponin in cell culture of Panax ginseng. Plant Cell Rep. 2001;20(7):674–677.

Wan L, Ma XJ, Lai J, Mo CM, Feng S, Luo H. Growth curve of Siraitia grosvenorii and correlative analysis of seed and growth of fruit. Zhongguo Zhong Yao Za Zhi. 2011;36(3):272–275.

Wang QJ, Zheng LP, Zhao PF, Zhao YL, Wang JW. Cloning and characterization of an elicitor-responsive gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase involved in 20-hydroxyecdysone production in cell cultures of Cyanotis arachnoidea. Plant Physiol Biochem. 2014;84:1–9.

Wang JR, Lin JF, Guo LQ, You LF, Zeng XL, Wen JM. Cloning and characterization of squalene synthase gene from Poria cocos and its up-regulation by methyl jasmonate. World J Microbiol Biotechnol. 2014;30(2):613–620.