Abstract
WRKY transcription factors play a critical role in metabolism and stress responses in plants, but few WRKYs have been reported in Taraxacum antungense. Here, a multiple stress-inducible gene, TaWRKY14, was isolated from T. antungense. TaWRKY14 was localized to the nuclei, and phylogenetic analysis indicated that TaWRKY14 shared the highest identity to WRKY40 in Salvia miltiorrhiza. TaWRKY14 was highly expressed in roots, and up-regulated by salt and drought stress, salicylic acid treatment and powdery mildew. TaWRKY14-overexpressing transgenic lines had higher chlorogenic acid concentration and high expression of TaPAL1. The yeast one-hybrid assay proved that TaWRKY14 bond to the W-box of proTaPAL1. Additionally, field experiments showed that TaWRKY14-overexpressing T. antungense lines have higher powdery mildew resistance than the wild-type. Our results demonstrated that TaWRKY14 can regulate CGA biosynthesis and play an important role in resistance to powdery mildew in T. antungense. TaWRKY14 transgenic T. antungense can be used for further evaluation as a new germplasm resource.
Key message
We analyzed and evaluated the biological function of TaWRKY14 in Taraxacum antungense, and determined that TaWRKY14-overexpressing lines have higher chlorogenic acid concentration and stronger resistance to powdery mildew.
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Abbreviations
- SA:
-
Salicylic acid
- TFs:
-
Transcription factors
- CGA:
-
Chlorogenic acid
- PAL:
-
Phenylalanine ammonia-lyase
- 4CL:
-
4-Coumarate-CoA ligase
- C4H:
-
Cinnamate 4-hydroxylase
- HCT:
-
Hydroxycinnamoyl-coenzyme (Co)A shikimate/quinate hydroxycinnamoyl transferase
- HQT:
-
Hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase
- HPLC:
-
High-performance liquid chromatography
- MS:
-
Murashige and Skoog
- qRT-PCR:
-
Quantitative real-time polymerase chain reaction
- MDA:
-
Malonic dialdehyde
- SOD:
-
Super oxide dismutase
- POD:
-
Peroxidase
- Y1H:
-
Yeast one-hybrid
References
Cao W, Wang Y, Shi M, Hao X, Zhao W, Wang Y, Kai G (2018) Transcription factor SmWRKY1 positively promotes the biosynthesis of Tanshinones in Salvia miltiorrhiza. Front Plant Sci 9:554. https://doi.org/10.3389/fpls.2018.00554
Chi Y, Yang Y, Zhou Y, Zhou J, Fan B, Yu JQ, Chen Z (2013) Protein-protein interactions in the regulation of WRKY transcription factors. Mol Plant 6(2):287–300. https://doi.org/10.1093/mp/sst026
Chhon S, Jeon J, Kim J, Park SU (2020) Accumulation of anthocyanins through overexpression of AtPAP1 in Solanum nigrum Lin. (Black Nightshade). Biomolecules 10(2):277. https://doi.org/10.3390/biom10020277
Deng C, Hao X, Shi M, Fu R, Wang Y, Zhang Y, Zhou W, Feng Y, Makunga N, Kai G (2019) Tanshinone production could be increased by the expression of SmWRKY2 in Salvia miltiorrhiza hairy roots. Plant Sci 284:1–8. https://doi.org/10.1016/j.plantsci.2019.03.007
Dewdney J, Reuber TL, Wildermuth MC, Devoto A, Cui J, Stutius LM, Drummond EP, Ausubel FM (2000) Three unique mutants of Arabidopsis identify eds loci required for limiting growth of a biotrophic fungal pathogen. Plant J 24:205–218. https://doi.org/10.1046/j.1365-313x.2000.00870.x
Fatemeh K, Houshang A, Mojtaba R (2018) Molecular cloning, functional characterization and expression of a drought inducible phenylalanine ammonia-lyase gene (ObPAL) from Ocimum basilicum L. Plant Physiol Biochem 130:464–472. https://doi.org/10.1016/j.plaphy.2018.07.026
Gao H, Chai H, Cheng N, Cao W (2017) Effects of 24-epibrassinolide on enzymatic browning and antioxidant activity of fresh-cut lotus root slices. Food Chem 217:45–51. https://doi.org/10.1016/j.foodchem.2016.08.063
Gharibia S, Tabatabaei EBS, Saeidi G, Talebib M, Matkowski A (2019) The effect of drought stress on polyphenolic compounds and expression of flavonoid biosynthesis related genes in Achillea pachycephala Rech.f. Phytochemistry 162:90–98. https://doi.org/10.1016/j.phytochem.2019.03.004
Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827. https://doi.org/10.1111/j.1365-313X.2007.03373.x
Grunewald W, Ive DS, Lewis DR, Löfke C, Jansen L, Goeminne G, Bossche RV, Karimi M, De Rybel B, Vanholme B (2012) Transcription factor WRKY23 assists auxin distribution patterns during Arabidopsis root development through local control on flavonol biosynthesis. PNAS 109:1554–1559. https://doi.org/10.1073/pnas.1121134109
Güneş A, Kordali S, Turan M, Bozhüyük AU (2019) Determination of antioxidant enzyme activity and phenolic contents of some species of the Asteraceae family from medicanal plants. Ind Crop Prod 137:208–213. https://doi.org/10.1016/j.indcrop.2019.05.042
Guo H, Wang HC, Xiang LG, Chen XJ, Meng JY, He YF, Ding W (2019) Powdery mildew of Taraxacum mongolicum caused by Podosphaera macrospora in China. Plant Dis 103:1420–1421. https://doi.org/10.1094/PDIS-10-18-1879-PDN
Hao X, Pu Z, Cao G, You D, Zhou Y, Deng C, Shi M, Nile SH, Wang Y, Zhou W, Kai G (2020) Tanshinone and salvianolic acid biosynthesis are regulated by SmMYB98 in Salvia miltiorrhiza hairy roots. J Adv Res 23:1–12. https://doi.org/10.1016/j.jare.2020.01.012
Huang Q, Sun M, Yuan T, Wang Y, Shi M (2019) The AP2/ERF transcription factor SmERF1L1 regulates the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza. Food Chem 274:368–375. https://doi.org/10.1016/j.foodchem.2018.08.119
Jędrejek D, Kontek B, Lis B, Stochmal A, Olas B (2017) Evaluation of antioxidant activity of phenolic fractions from the leaves and petals of dandelion in human plasma treated with H2O2 and H2O2/Fe. Chem-Biol Interact 262:29–37. https://doi.org/10.1016/j.cbi.2016.12.003
Jing J, Zhang H, Xue Y, Zeng K (2020) Effects of INA on postharvest blue and green molds and anthracnose decay in citrus fruit. J Integr Agr 19(5):1396–1406. https://doi.org/10.1016/S2095-3119(20)63169-0
Kagale S, Links MG, Rozwadowski K (2010) Genome-wide analysis of ethylene-responsive element binding factor-associated amphiphilic repression motif-containing transcriptional regulators in Arabidopsis. Plant Physiol 152:1109–1134. https://doi.org/10.4161/psb.5.6.11576
Kang J, Guo Y, Chen Y, Li H, Zhang L, Liu H (2014) Upregulation of the AT-hook DNA binding gene BoMF2 in OguCMS anthers of Brassica oleracea suggests that it encodes a transcriptional regulatory factor for anther development. Mol Biol Rep 41:2005–2014. https://doi.org/10.1007/s11033-014-3048-2
Karagöezler AA, Erdag B, Emek YC, Uygun DA (2008) Antioxidant activity and proline content of leaf extracts from Dorystoechas hastata. Food Chem 111:400–407. https://doi.org/10.1016/j.foodchem.2008.03.089
Khakdan F, Alizadeh H, Ranjbar M (2018) Molecular cloning, functional characterization and expression of a drought inducible phenylalanine ammonia-lyase gene (ObPAL) from Ocimum basilicum L. Plant Physiol Biochem 130:464–472. https://doi.org/10.1016/j.plaphy.2018.07.026
Kim DS, Hwang BK (2014) An important role of the pepper phenylalanine ammonia-lyase gene (PAL1) in salicylic acid-dependent signalling of the defence response to microbial pathogens. J Exp Bot 65:2295–2306. https://doi.org/10.1093/jxb/eru109
Li S, Fu Q, Chen L, Huang W, Yu D (2011) Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance. Planta 233:1237–1252. https://doi.org/10.1007/s00425-011-1375-2
Li C, Li D, Shao F, Lu S (2015) Molecular cloning and expression analysis of WRKY transcription factor genes in Salvia miltiorrhiza. BMC Genomics 16:200. https://doi.org/10.1186/s12864-015-1411-x
Li X, Guo W, Li J, Yue P, Bu H, Jiang J, Liu W, Xu Y, Yuan H, Li T, Wang A (2020) Histone acetylation at the promoter for the transcription factor PuWRKY31 affects sucrose accumulation in pear fruit. Plant Physiol 182(4):2035–2046. https://doi.org/10.1104/pp.20.00002
Lin T, Xu X, Ruan J, Liu S, Wu S, Shao X, Wang X, Gan L, Qin B, Yang Y (2017) Genome analysis of Taraxacum kok-saghyz Rodin provides new insights into rubber biosynthesis. Natl Sci Rev 5:78–87. https://doi.org/10.1093/nsr/nwx101
Lis B, Olas B (2019) Pro-health activity of dandelion (Taraxacum officinale L.) and its food products–history and present. J Funct Foods 50:40–48. https://doi.org/10.1016/j.jff.2019.05.012
Liu XQ, Bai XQ, Qian Q, Wang XJ, Chen MS, Chu CC (2005) OsWRKY03, a rice transcriptional activator that functions in defense signaling pathway upstream of OsNPR1. Cell Res 15(8):593–603. https://doi.org/10.1038/sj.cr.7290329
Liu Q, Liu Y, Xu Y, Yao L, Liu Z, Cheng H, Ma M, Wu J, Wang W, Ning W (2018) Overexpression of and RNA interference with hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase affect the chlorogenic acid metabolic pathway and enhance salt tolerance in Taraxacum antungense Kitag. Phytochem Lett 28:116–123. https://doi.org/10.1016/j.phytol.2018.10.003
Liu Q, Yao L, Xu Y, Cheng H, Wang W, Liu Z, Liu J, Cui X, Zhou Y, Ning W (2019) In vitro evaluation of hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase expression and regulation in Taraxacum antungense in relation to 5-caffeoylquinic acid production. Phytochemistry 162:148–156. https://doi.org/10.1016/j.phytochem.2019.02.014
Martinez M, Poirrier P, Chamy R, Prüfer D, Schulze-Gronover C, Jorquera L, Ruiz G (2015) Taraxacum officinale and related species—an ethnopharmacological review and its potential as a commercial medicinal plant. J Ethnopharmacol 169:244–262. https://doi.org/10.1016/j.jep.2015.03.067
Niggeweg R, Michael AJ, Martin C (2004) Engineering plants with increased levels of the antioxidant chlorogenic acid. Nat Biotechnol 22(6):746–754. https://doi.org/10.1038/nbt966
Noman M, Jameel A, Qiang WD, Ahmad N, Liu WC, Wang FW, Li HY (2019) Overexpression of GmCAMTA12 enhanced drought tolerance in Arabidopsis and Soybean. Int J Mol Sci 20(19):4849. https://doi.org/10.3390/ijms20194849
Pandey N, Goswami N, Tripathi D, Rai KK, Rai SK, Singh S, Pandey-Rai S (2019) Epigenetic control of UV-B-induced flavonoid accumulation in Artemisia annua L. Planta 249(2):497–514. https://doi.org/10.1007/s00425-018-3022-7
Patel JS, Selvaraj V, Gunupuru LR, Rathor PK, Prithiviraj B (2020) Combined application of Ascophyllum nodosum extract and chitosan synergistically activates host-defense of peas against powdery mildew. BMC Plant Biol 20(1):113. https://doi.org/10.1186/s12870-020-2287-8
Qiu D, Xie X, Wei B, Liu H, Li X, Xiong L, Wang S (2008) Rice gene network inferred from expression profiling of plants overexpressing OsWRKY13, a positive regulator of disease resistance. Mol Plant 1:538–551. https://doi.org/10.1093/mp/ssn012
Rahman A, Wallis CM, Uddin W (2015) Silicon-induced systemic defense responses in Perennial Ryegrass against infection by Magnaporthe oryzae. Phytopathology 105:748–757. https://doi.org/10.1094/PHYTO-12-14-0378-R
Shi M, Liao P, Nile SH, Georgiev MI, Kai G (2020) Biotechnological exploration of transformed root culture for value-added products. Trends Biotechnol. https://doi.org/10.1016/j.tibtech.2020.06.012
Shine MB, Yang JW, El-Habbak M, Nagyabhyru P, Fu DQ, Navarre D, Ghabrial S, Kachroo P, Kachroo A (2016) Cooperative functioning between phenylalanine ammonia lyase and isochorismate synthase activities contributes to salicylic acid biosynthesis in soybean. New Phytol 212:627–636. https://doi.org/10.1111/nph.14078
Sun PW, Xu YH, Yu CC, Lv FF, Tang X, Gao ZH, Zhang Z, Wang H, Liu Y, Wei JH (2020) WRKY44 represses expression of the wound-induced sesquiterpene biosynthetic gene ASS1 in Aquilaria sinensis. J Exp Bot 71(3):1128–1138. https://doi.org/10.1093/jxb/erz469
Sziderics AH, Oufir M, Trognitz F, Kopecky D, Matusíková I, Hausman JF, Wilhelm E (2010) Organ-specific defence strategies of pepper (Capsicum annuum L.) during early phase of water deficit. Plant Cell Rep 29:295–305. https://doi.org/10.1007/s00299-010-0822-z
Tian X, Zhang L, Feng S, Zhao Z, Wang X, Gao H (2019) Transcriptome analysis of apple leaves in response to powdery mildew (Podosphaera leucotricha) infection. Int J Mol Sci 20(9):2326. https://doi.org/10.3390/ijms20092326
Wang M, Vannozzi A, Wang G, Liang YH, Tornielli GB, Zenoni S, Cavallini E, Pezzotti M, Cheng ZM (2014) Genome and transcriptome analysis of the grapevine (Vitis vinifera L.) WRKY gene family. Hortic Res 1:16. https://doi.org/10.1038/hortres.2014.16
Wang C, Wu C, Wang Y, Xie C, Shi M, Nile S, Zhou Z, Kai G (2019) Transcription factor OpWRKY3 is involved in the development and biosynthesis of camptothecin and its precursors in Ophiorrhiza pumila hairy roots. Int J Mol Sci. https://doi.org/10.3390/ijms20163996
Xu M, Wu C, Zhao LM, Wang Y, Wang C, Zhou W, Ming Y, Kai G (2020) WRKY transcription factor OpWRKY1 acts as a negative regulator of camptothecin biosynthesis in Ophiorrhiza pumila hairy roots. Plant Cell Tissue Organ Cult 142(1):69–78. https://doi.org/10.1007/s11240-020-01833-2
Yoshikawa M, Luo W, Tanaka G, KonishiY MH (2018) Wounding stress induces phenylalanine ammonia lyases, leading to the accumulation of phenylpropanoids in the model liverwort Marchantia polymorpha. Phytochemistry 155:30–36. https://doi.org/10.1016/j.phytochem.2018.07.014
Yu D, Chen C, Chen Z (2001) Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13:1527–1539. https://doi.org/10.1105/tpc.13.7.1527
Yuan W, Jiang T, Du K, Chen H, Cao Y, Xie J, Li M, Carr JP, Wu B, Fan Z, Zhou T (2019) Maize phenylalanine ammonia-lyases contribute to resistance to Sugarcane mosaic virus infection, mostlikely throughpositive regulation of salicylic acid accumulation. Mol Plant Pathol 20:1365–1378. https://doi.org/10.1111/mpp.12817
Zhang J, Yang Y, Zheng K, Xie M, Feng K, Jawdy SS, Gunter LE, Ranjan P, Singan VR, Engle N, Lindquist E, Barry K, Schmutz J, Zhao N, Tschaplinski TJ, LeBoldus J, Tuskan G, Chen JC, Muchero W (2018a) Genome-wide association studies and expression-based quantitative trait loci analyses reveal roles of HCT2 in caffeoylquinic acid biosynthesis and its regulation by defense-responsive transcription factors in Populus. New Phytol 220:502–516. https://doi.org/10.1111/nph.15297
Zhang L, Cheng J, Sun X, Zhao T, Li M (2018b) Overexpression of VaWRKY14 increases drought tolerance in Arabidopsis by modulating the expression of stress-related genes. Plant Cell Rep 37:1159–1172. https://doi.org/10.1007/s00299-018-2302-9
Zhou W, Qian C, Lia R, Zhou S, Zhang R, Xiao J, Wang X, Zhang S, Xing L, Ca A (2018) TaNAC6s are involved in the basal and broad-spectrum resistance to powdery mildew in wheat. Plant Sci 277:218–228. https://doi.org/10.1016/j.plantsci.2018.09.014
Acknowledgements
This work was supported by National Natural Science Fund of China [Grant Numbers: 82073963, 81522049, 81703636, 31571735, 31270007]; Zhejiang Provincial Ten Thousand Program for Leading Talents of Science and Technology Innovation [2018R52050]; Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents. The Opening Project of Zhejiang Provincial Preponderant and Characteristic Subject of Key University (Traditional Chinese Pharmacology), Zhejiang Chinese Medical University [ZYAOX2018004]. Funds were used for collection and analysis of data in this study, as well as in the open access payment. We would like to thank Editage (www.editage.cn) for English language editing.
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LQ designed and planned the experiments and wrote the paper. LQ, ZW, CH, WJ, and WL, collected the materials. LQ, RQ, LT, YY, JJ, and LL performed the experiments. LQ and ZW compiled and interpreted the data. LQ wrote the manuscript. Prof. KG, NW and JJ revised the manuscript.
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Qun Liu and Wei Zhou have contributed equally to this work.
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Liu, Q., Zhou, W., Ruan, Q. et al. Overexpression of TaWRKY14 transcription factor enhances accumulation of chlorogenic acid in Taraxacum antungense Kitag and increases its resistance to powdery mildew. Plant Cell Tiss Organ Cult 143, 665–679 (2020). https://doi.org/10.1007/s11240-020-01950-y
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DOI: https://doi.org/10.1007/s11240-020-01950-y