Abstract
Liquorice (Glycyrrhiza uralensis) is an important medicinal plant for which there is a huge market demand. It has been reported that arbuscular mycorrhizal (AM) symbiosis and drought stress can stimulate the accumulation of the active ingredients, glycyrrhizin and liquiritin, in liquorice plants, but the potential interactions of AM symbiosis and drought stress remain largely unknown. In the present work, we investigated mycorrhizal effects on plant growth and accumulation of glycyrrhizin and liquiritin in liquorice plants under different water regimes. The results indicated that AM plants generally exhibited better growth and physiological status including stomatal conductance, photosynthesis rate, and water use efficiency compared with non-AM plants. AM inoculation up-regulated the expression of an aquaporin gene PIP and decreased root abscisic acid (ABA) concentrations under drought stress. In general, AM plants displayed lower root carbon (C) and nitrogen (N) concentrations, higher phosphorus (P) concentrations, and therefore, lower C:P and N:P ratios but higher C:N ratio than non-AM plants. On the other hand, AM inoculation increased root glycyrrhizin and liquiritin concentrations, and the mycorrhizal effects were more pronounced under moderate drought stress than under well-watered condition or severe drought stress for glycyrrhizin accumulation. The accumulation of glycyrrhizin and liquiritin in AM plants was consistent with the C:N ratio changes in support of the carbon-nutrient balance hypothesis. Moreover, the glycyrrhizin accumulation was positively correlated with the expression of glycyrrhizin biosynthesis genes SQS1, β-AS, CYP88D6, and CYP72A154. By contrast, no significant interaction of AM inoculation with water treatment was observed for liquiritin accumulation, while we similarly observed a positive correlation between liquiritin accumulation and the expression of a liquiritin biosynthesis gene CHS. These results suggested that AM inoculation in combination with proper water management potentially could improve glycyrrhizin and liquiritin accumulation in liquorice roots and may be practiced to promote liquorice cultivation.
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Al-Karaki G, McMichael B, Zak J (2004) Field response of wheat to arbuscular mycorrhizal fungi and drought stress. Mycorrhiza 14:263–269
Aquil S, Husaini AM, Abdin MZ, Rather GM (2009) Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants. Planta Med 75:1453–1458
Aroca R, Vernieri P, Ruiz-Lozano JM (2008) Mycorrhizal and non-mycorrhizal Lactuca sativa plants exhibit contrasting responses to exogenous ABA during drought stress and recovery. J Exp Bot 59:2029–2041
Babikova Z, Gilbert L, Bruce T, Dewhirst SY, Pickett JA, Johnson D (2014) Arbuscular mycorrhizal fungi and aphids interact by changing host plant quality and volatile emission. Funct Ecol 28:375–385
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207
Box GEP, Cox DR (1964) An analysis of transformations. J R Stat Soc Ser B Methodol 2:211–252
Bryant JP, Chapin FS, Klein DR (1983) Carbon/nutrient balance of boreal plants in relation to vertebrate herbivory. Oikos 40:357–368
Chappell J, Wolf F, Proulx J, Cuellar R, Saunders C (1995) Is the reaction catalyzed by 3-hydroxy-3-methylglutaryl coenzyme a reductase a rate-limiting step for isoprenoid biosynthesis in plants? Plant Physiol 109:1337–1343
Chen MM, Yin HB, O’Connor P, Wang YS, Zhu YG (2010) C: N: P stoichiometry and specific growth rate of clover colonized by arbuscular mycorrhizal fungi. Plant Soil 326:21–29
Chen ML, Yang G, Sheng Y, Li PY, Qiu HY, Zhou XT, Huang LQ, Chao Z (2017) Glomus mosseae inoculation improves the root system architecture, photosynthetic efficiency and flavonoids accumulation of liquorice under nutrient stress. Front Plant Sci 8:931
Chitarra W, Pagliarani C, Maserti B, Lumini E, Siciliano I, Cascone P, Schubert A, Gambino G, Balestrini R, Guerrieri E (2016) Insights on the impact of arbuscular mycorrhizal symbiosis on tomato tolerance to water stress. Plant Physiol 171:1009–1023
Dai ZB, Cui GH, Zhou SF, Zhang XA, Huang LQ (2011) 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
de Abreu IN, Mazzafera P (2005) Effect of water and temperature stress on the content of active constituents of Hypericum brasiliense Choisy. Plant Physiol Biochem 43:241–248
Elser JJ, Dobberfuhl DR, MacKay NA, Schampel JH (1996) Organism size, life history, and N: P stoichiometry. Bioscience 46:674–684
Enfissi E, Fraser PD, Lois LM, Boronat A, Schuch W, Bramley PM (2005) Metabolic engineering of the mevalonate and non-mevalonate isopentenyl diphosphate-forming pathways for the production of health-promoting isoprenoids in tomato. Plant Biotechnol J 3:17–27
Gianinazzi-Pearson V (1996) Plant cell responses to arbuscular mycorrhizal fungi: getting to the roots of the symbiosis. Plant Cell 8:1871–1883
Golldack D, Li C, Mohan H, Probst N (2014) Tolerance to drought and salt stress in plants: unraveling the signaling networks. Front Plant Sci 5:151
Gorelick J, Bernstein N (2014) Elicitation: an underutilized tool for the development of medicinal plants as a source for therapeutic secondary metabolites. Adv Agron 124:201–230
Hamilton JG, Zangerl AR, DeLucia EH, Berenbaum MR (2001) The carbon-nutrient balance hypothesis: its rise and fall. Ecol Lett 4:86–95
Harker M, Holmberg N, Clayton JC, Gibbard CL, Wallace AD, Rawlins S, Hellyer SA, Lanot A, Safford R (2003) Enhancement of seed phytosterol levels by expression of an N-terminal truncated Hevea brasiliensis (rubber tree) 3-hydroxy-3-methylglutaryl-CoA reductase. Plant Biotechnol J 1:113–121
Hayashi H (2009) Molecular biology of secondary metabolism: case study for Glycyrrhiza plants. In: Recent Advances in Plant Biotechnology. Springer, Boston, pp 89–103
Hayashi H, Sudo H (2009) Economic importance of licorice. Plant Biotechnol 26:101–104
He MZ, Dijkstra FA (2014) Drought effect on plant nitrogen and phosphorus: a meta-analysis. New Phytol 204:924–931
Helber N, Wippel K, Sauer N, Schaarschmidt S, Hause B, Requena N (2011) A versatile monosaccharide transporter that operates in the arbuscular mycorrhizal fungus Glomus sp is crucial for the symbiotic relationship with plants. Plant Cell 23:3812–3823
Hong JH, Seah SW, Xu J (2013) The root of ABA action in environmental stress response. Plant Cell Rep 32:971–983
Javot H, Maurel C (2002) The role of aquaporins in root water uptake. Ann Bot 90:301–313
Kapoor R, Chaudhary V, Bhatnagar AK (2007) Effects of arbuscular mycorrhiza and phosphorus application on artemisinin concentration in Artemisia annua L. Mycorrhiza 17:581–587
Kapoor R, Anand G, Gupta P, Mandal S (2017) Insight into the mechanisms of enhanced production of valuable terpenoids by arbuscular mycorrhiza. Phytochem Rev 16:677–692
Kim YJ, Lee CS (2008) Glycyrrhizin attenuates MPTP neurotoxicity in mouse and MPP+-induced cell death in PC12 cells. Korean J Physiol Pharmacol 12:65–71
Kim YS, Cho JH, Ahn J, Hwang B (2006) Upregulation of isoprenoid pathway genes during enhanced saikosaponin biosynthesis in the hairy roots of Bupleurum falcatum. Mol Cells 22:269–274
Kitagawa I (2002) Licorice root. A natural sweetener and an important ingredient in Chinese medicine. Pure Appl Chem 74:1189–1198
Laule O, Fürholz A, Chang HS, Zhu T, Wang X, Heifetz PB, Gruissem W, Lange BM (2003) Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. Proc Natl Acad Sci 100:6866–6871
Li WD, Hou JL, Wang WQ, Tang XM, Liu CL, Xing D (2011) Effect of water deficit on biomass production and accumulation of secondary metabolites in roots of Glycyrrhiza uralensis. Russ J Plant Physiol 58:538–542
Li T, Hu YJ, Hao ZP, Li H, Wang YS, Chen BD (2013) First cloning and characterization of two functional aquaporin genes from an arbuscular mycorrhizal fungus Glomus intraradices. New Phytol 197:617–630
Li T, Sun YQ, Ruan Y, Xu LJ, Hu YJ, Hao ZP, Zhang X, Li H, Wang YS, Yang LG, Chen BD (2016a) Potential role of D-myo-inositol-3-phosphate synthase and 14-3-3 genes in the crosstalk between Zea mays and Rhizophagus intraradices under drought stress. Mycorrhiza 26:879–893
Li YP, Yu CX, Qiao J, Zang YM, Xiang Y, Ren GX, Wang L, Zhang XY, Liu CS (2016b) Effect of exogenous phytohormones treatment on glycyrrhizic acid accumulation and preliminary exploration of the chemical control network based on glycyrrhizic acid in root of Glycyrrhiza uralensis. Rev Bras 26:490–496
Li JL, Liu SJ, Wang J, Li J, Li JX, Gao WY (2017) Gene expression of glycyrrhizin acid and accumulation of endogenous signaling molecule in Glycyrrhiza uralensis Fisch adventitious roots after Saccharomyces cerevisiae and Meyerozyma guilliermondii applications. Biotechnol Appl Biochem 64:700–711
Liu Y, Liu CS (2012) Study on the spatial and temporal expression of beta-AS gene of Glycyrriza uralensis. J Chin Med Mater 35:528–531
Liu JN, Wu LJ, Wei SL, Xiao X, Sun CX, Jiang P, Song ZB, Wang T, Yu ZL (2007) Effects of arbuscular mycorrhizal fungi on the growth, nutrient uptake and glycyrrhizin production of licorice (Glycyrrhiza uralensis Fisch). Plant Growth Regul 52:29–39
Liu HY, Wang XD, Wang DH, Zou ZR, Liang ZS (2011) Effect of drought stress on growth and accumulation of active constituents in Salvia miltiorrhiza Bunge. Ind Crop Prod 33:84–88
Liu Y, Xu QX, Xi PY, Chen HH, Liu CS (2013) Cloning and characterization of a cDNA coding 3-hydroxy-3-methylglutary CoA reductase involved in glycyrrhizic acid biosynthesis in Glycyrrhiza uralensis. Acta Pharm Sin 48:773–779
López-Ráez JA, Flors V, García JM, Pozo MJ (2010) AM symbiosis alters phenolic acid content in tomato roots. Plant Signal Behav 5:1138–1140
Mandal S, Upadhyay S, Wajid S, Ram M, Jain DC, Singh VP, Abdin MZ, Kapoor R (2015) Arbuscular mycorrhiza increase artemisinin accumulation in Artemisia annua by higher expression of key biosynthesis genes via enhanced jasmonic acid levels. Mycorrhiza 25:345–357
Mansouri H, Asrar Z, Szopa J (2009) Effects of ABA on primary terpenoids and ∆9-tetrahydrocannabinol in Cannabis sativa L. at flowering stage. Plant Growth Regul 58:269–277
Matzek V, Vitousek PM (2009) N: P stoichiometry and protein: RNA ratios in vascular plants: an evaluation of the growth-rate hypothesis. Ecol Lett 12:765–771
Mochida K, Sakurai T, Seki H, Yoshida T, Takahagi K, Sawai S, Uchiyama H, Muranaka T, Saito K (2017) Draft genome assembly and annotation of Glycyrrhiza uralensis, a medicinal legume. Plant J 89:181–194
Nasrollahi V, Mirzaie-asl A, Piri K, Nazeri S, Mehrabi R (2014) The effect of drought stress on the expression of key genes involved in the biosynthesis of triterpenoid saponins in liquorice (Glycyrrhiza glabra). Phytochemistry 103:32–37
Neumann E, Schmid B, Römheld V, George E (2009) Extraradical development and contribution to plant performance of an arbuscular mycorrhizal symbiosis exposed to complete or partial rootzone drying. Mycorrhiza 20:13–23
Nowak M, Kleinwächter M, Manderscheid R, Weigel HJ, Selmar D (2010) Drought stress increases the accumulation of monoterpenes in sage (Salvia officinalis), an effect that is compensated by elevated carbon dioxide concentration. J Appl Bot Food Qual 83:133–136
Okubo K, Yoshiki Y (2000) The role of triterpenoid on reactive oxygen scavenging system: approach from the new chemiluminescence system (XYZ system). Biofactors 13:219–223
Orujei Y, Shabani L, Sharifi-Tehrani M (2013) Induction of glycyrrhizin and total phenolic compound production in licorice by using arbuscular mycorrhizal fungi. Russ J Plant Physiol 60:855–860
Pan Y, Wu LJ, Yu ZL (2006) Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhiza uralensis Fisch). Plant Growth Regul 49:157–165
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:45
Porcel R, Azcón R, Ruiz-Lozano JM (2004) Evaluation of the role of genes encoding for Δ1-pyrroline-5-carboxylate synthetase (P5CS) during drought stress in arbuscular mycorrhizal Glycine max and Lactuca sativa plants. Physiol Mol Plant Pathol 65:211–221
Rodríguez-Concepción M (2006) Early steps in isoprenoid biosynthesis: multilevel regulation of the supply of common precursors in plant cells. Phytochem Rev 5:1–15
Ruiz-Lozano JM, Aroca R, Zamarreño ÁM, Molina S, Andreo-Jiménez B, Porcel R, García-Mina JM, Ruyter-Spira C, López-Ráez JA (2016) Arbuscular mycorrhizal symbiosis induces strigolactone biosynthesis under drought and improves drought tolerance in lettuce and tomato. Plant Cell Environ 39:441–452
Schenke D, Böttcher C, Scheel D (2011) Crosstalk between abiotic ultraviolet-B stress and biotic (flg22) stress signalling in Arabidopsis prevents flavonol accumulation in favor of pathogen defence compound production. Plant Cell Environ 34:1849–1864
Seki H, Ohyama K, Sawai S, Mizutani M, Ohnishi T, Sudo H, Akashi T, Aoki T, Saito K, Muranaka T (2008) Licorice β-amyrin 11-oxidase, a cytochrome P450 with a key role in the biosynthesis of the triterpene sweetener glycyrrhizin. Proc Natl Acad Sci 105:14204–14209
Seki H, Sawai S, Ohyama K, Mizutani M, Ohnishi T, Sudo H, Fukushima EO, Akashi T, Aoki T, Saito K (2011) Triterpene functional genomics in licorice for identification of CYP72A154 involved in the biosynthesis of glycyrrhizin. Plant Cell 23:4112–4123
Selmar D, Kleinwächter M (2013) Influencing the product quality by deliberately applying drought stress during the cultivation of medicinal plants. Ind Crop Prod 42:558–566
Shen ZY, Liu CS, Wang XY (2009) Cloning and characterization of open reading frame encoding beta-amyrin synthase in Glycyrrhiza uralensis. China J Chinese Materia Medica 34:2438–2440
Shrivastava G, Ownley BH, Augé RM, Toler H, Dee M, Vu A, Köllner TG, Chen F (2015) Colonization by arbuscular mycorrhizal and endophytic fungi enhanced terpene production in tomato plants and their defense against a herbivorous insect. Symbiosis 65:65–74
Sivakumaran A, Akinyemi A, Mandon J, Cristescu SM, Hall MA, Harren FJM, Mur LAJ (2016) ABA suppresses Botrytis cinerea elicited NO production in tomato to influence H2O2 generation and increase host susceptibility. Front Plant Sci 7:709
Smith SE, Smith FA, Jakobsen I (2003) Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses. Plant Physiol 133:16–20
Sun XG, Shi J, Ding GJ (2017) Combined effects of arbuscular mycorrhiza and drought stress on plant growth and mortality of forage sorghum. Appl Soil Ecol 119:384–391
Szabados L, Savouré A (2010) Proline: a multifunctional amino acid. Trends Plant Sci 15:89–97
Trouvelot A, Kough JL, Gianinazzi-Pearson V (1986) Mesure du taux de mycorhization VA d’un système radiculaire. Recherche de methods d’estimation ayant une signification fonctionnelle. In: Gianinazzi-Pearson V, Gianinazzi S (eds) Physiological and genetical aspects of mycorrhizae. INRA-Press, Paris, pp 217–221
Wei SL, Wang WQ, Wang H (2003) Study on licorice resources and their sustainable utilization in center and western area of China. China J Chinese Materia Medica 28:202–206
Welling MT, Liu L, Rose TJ, Waters DLE, Benkendorff K (2016) Arbuscular mycorrhizal fungi: effects on plant terpenoid accumulation. Plant Biol 18:552–562
Winkel-Shirley B (2002) Biosynthesis of flavonoids and effects of stress. Curr Opin Plant Biol 5:218–223
Xu GJ, Cai W, Gao W, Liu CS (2016) A novel glucuronosyltransferase has an unprecedented ability to catalyse continuous two-step glucuronosylation of glycyrrhetinic acid to yield glycyrrhizin. New Phytol 212:123–135
Yang DF, Sheng DF, Duan QM, Liang X, Liang ZS, Liu Y (2012) PEG and ABA trigger the burst of reactive oxygen species to increase tanshinone production in Salvia miltiorrhiza hairy roots. J Plant Growth Regul 31:579–587
Zeng Y, Guo LP, Chen BD, Hao ZP, Wang JY, Huang LQ, Yang G, Cui XM, Yang L, Wu ZX (2013) Arbuscular mycorrhizal symbiosis and active ingredients of medicinal plants: current research status and prospectives. Mycorrhiza 23:253–265
Zhang HC, Liu JM, Lu HY, Gao SL (2009) Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment. Plant Cell Rep 28:1205–1213
Zhang RQ, Zhu HH, Zhao HQ, Yao Q (2013) Arbuscular mycorrhizal fungal inoculation increases phenolic synthesis in clover roots via hydrogen peroxide, salicylic acid and nitric oxide signaling pathways. J Plant Physiol 170:74–79
Zhang Y, Zhou ZY, Xia PG, Liang ZS, Liu SL, Liu ZL (2016) Expression of key enzyme genes and content of saikosaponin in saikosaponin biosynthesis under drought stress in Bupleurum chinense. China J Chinese Materia Medica 41:643–647
Zhao RX, Guo W, Bi N, Guo JY, Wang LX, Zhao J, Zhang J (2015) Arbuscular mycorrhizal fungi affect the growth, nutrient uptake and water status of maize (Zea mays L.) grown in two types of coal mine spoils under drought stress. Appl Soil Ecol 88:41–49
Acknowledgements
We thank Dr. Yan Zeng from China National Traditional Chinese Medicine Corporation for kindly supplying the seeds of Glycyrrhiza uralensis.
Funding
This study was financially supported by the National Natural Science Foundation of China (project no. 41571250 and 41371264) and National Key Research and Development Program of China (project no. 2016YFC0500702).
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Xie, W., Hao, Z., Zhou, X. et al. Arbuscular mycorrhiza facilitates the accumulation of glycyrrhizin and liquiritin in Glycyrrhiza uralensis under drought stress. Mycorrhiza 28, 285–300 (2018). https://doi.org/10.1007/s00572-018-0827-y
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DOI: https://doi.org/10.1007/s00572-018-0827-y