Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 2, pp 249–259 | Cite as

Stress associated protein 1 regulates the development of glandular trichomes in Artemisia annua

  • Yuting Wang
  • Xueqing Fu
  • Lihui Xie
  • Wei Qin
  • Ling Li
  • Xiaofen Sun
  • Shihai XingEmail author
  • Kexuan TangEmail author
Original Article


Artemisinin, a sesquiterpene lactone endoperoxide extracted from the aerial part of Artemisia annua L., is widely known as the useful treatment for malaria. Increasing the density of the glandular trichomes in A. annua is an effective method for increasing the production of artemisinin. Here, we identified a transcription factor, AaSAP1 containing A20/AN1 zinc finger motif, which encodes stress associated protein 1 (SAP1). The expression analysis in various tissues indicated that AaSAP1 predominately expressed in the trichomes. Methyl jasmonate, abscisic acid and gibberellic acid induced the expression of AaSAP1. Notably, up-regulation or down-regulation of the transcriptional level of AaSAP1 led to an increase or decrease in the density of the glandular trichomes of A. annua, respectively. In addition, overexpression of AaSAP1 significantly enhanced the content of artemisinin in A. annua. Our results reveal that AaSAP1 positively regulates the development of the glandular trichomes, and it is a valuable gene in genetic engineering of A. annua for increasing the production of artemisinin.

Key message

AaSAP1 positively regulates the development of the glandular trichomes and the production of artemisinin in A. annua.


Artemisia annua Stress associated protein Glandular trichome Artemisinin 



This research was funded by Grants from the Bill & Melinda Gates Foundation (OPP1199872); the National Science Foundation of China (18Z103150043); China National Key Research and Development Program (2017ZX09101002-003-002).

Author contributions

YW and KT conceived and designed the project. YW, XF, LX and WQ conducted the experiments. YW, FX, LL and XS analyzed the data. YW and XF drafted the paper. KT and SX reviewed the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Human or animal rights

This article does not contain any studies with human or animal subjects performed by the any of the authors.


  1. Berg JM, Shi Y (1996) The galvanization of biology: a growing appreciation for the roles of zinc. Science 271:1081–1085. CrossRefPubMedGoogle Scholar
  2. Charrier A, Planchet E, Cerveau D, Gimeno-Gilles C, Verdu I, Limami AM, Lelièvre E (2012) Overexpression of a Medicago truncatula stress-associated protein gene (MtSAP1) leads to nitric oxide accumulation and confers osmotic and salt stress tolerance in transgenic tobacco. Planta 236:567–577. CrossRefPubMedGoogle Scholar
  3. Duke SO, Paul RN (1993) Development and fine structure of the glandular trichomes of Artemisia annua L. Int J Plant Sci 154:107–118. CrossRefGoogle Scholar
  4. Efferth T (2006) Molecular pharmacology and pharmacogenomics of artemisinin and its derivatives in cancer cells. Curr Drug Targets 7:407–421. CrossRefPubMedGoogle Scholar
  5. Fu XQ, Shi P, He Q et al (2017) AaPDR3, a PDR transporter 3, is involved in sesquiterpene β-caryophyllene transport in Artemisia annua. Front Plant Sci 8:723. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Giri J, Vij S, Dansana PK, Tyagi AK (2011) Rice A20/AN1 zinc-finger containing stress-associated proteins (SAP1/11) and a receptor-like cytoplasmic kinase (OsRLCK253) interact via A20 zinc-finger and confer abiotic stress tolerance in transgenic Arabidopsis plants. New Phytol 191:721–732. CrossRefPubMedGoogle Scholar
  7. Giri J, Dansana PK, Kothari KS, Sharma G, Vij S, Tyagi AK (2013) SAPs as novel regulators of abiotic stress response in plants. BioEssays 35:639–648. CrossRefPubMedGoogle Scholar
  8. Graham IA, Besser K, Blumer S et al (2010) The genetic map of Artemisia annua L. identifies loci affecting yield of the antimalarial drug artemisinin. Science 327:328–331. CrossRefPubMedGoogle Scholar
  9. He Q, Fu XQ, Shi P, Liu M, Shen Q, Tang KX (2017) Glandular trichome-specific expression of alcohol dehydrogenase 1 (ADH1) using a promoter-GUS fusion in Artemisia annua L. Plant Cell Tissue Org Cult 130:61–72. CrossRefGoogle Scholar
  10. Hong GJ, Xue XY, Mao YB, Wang LJ, Chen XY (2012) Arabidopsis MYC2 interacts with DELLA proteins in regulating sesquiterpene synthase gene expression. Plant Cell 24(6):2635–2648. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Hülskamp M (2004) Plant trichomes: a model for cell differentiation. Nat Rev Mol Cell Biol 5:471–480. CrossRefPubMedGoogle Scholar
  12. Jenkins TH, Li J, Scutt CP, Gilmartin PM (2005) Analysis of members of the Silene latifolia Cys2/His2 zinc-finger transcription factor family during dioecious flower development and in a novel stamen-defective mutant ssf1. Planta 220:559–571. CrossRefPubMedGoogle Scholar
  13. Jiang WM, Lu X, Bo Q et al (2014) Molecular cloning and characterization of a trichome-specific promoter of artemisinic aldehyde Δ11(13) reductase (DBR2) in Artemisia annua. Plant Mol Biol Rep 32:82–91. CrossRefGoogle Scholar
  14. Jin Y, Wang M, Fu JJ et al (2007) Phylogenetic and expression analysis of ZnF-AN1 genes in plants. Genomics 90:265–275. CrossRefPubMedGoogle Scholar
  15. Johnson HB (1975) Plant pubescence: an ecological perspective. Bot Rev 41:233–258. CrossRefGoogle Scholar
  16. Kang M, Fokar M, Abdelmageed H, Allen RD (2011) Arabidopsis SAP5 functions as a positive regulator of stress responses and exhibits E3 ubiquitin ligase activity. Plant Mol Biol 75:451–466. CrossRefPubMedGoogle Scholar
  17. Kang M, Lee S, Abdelmagged H et al (2017) Arabidopsis stress associated protein 9 mediates biotic and abiotic stress responsive ABA signaling via the proteasome pathway. Plant Cell Environ 40:702–716. CrossRefPubMedGoogle Scholar
  18. Kumar S, Gupta SK, Singh P et al (2004) High yields of artemisinin by multi-harvest of Artemisia annua crops. Ind Crop Prod 19:77–90. CrossRefGoogle Scholar
  19. Liu YJ, Xu YY, Xiao J, Ma QB, Li D, Xue Z, Chong K (2011) OsDOG, a gibberellin-induced A20/AN1 zinc-finger protein, negatively regulates gibberellin-mediated cell elongation in rice. J Plant Physiol 168:1098–1105. CrossRefPubMedGoogle Scholar
  20. Liu M, Shi P, Fu XQ, Brodelius PE, Shen Q, Jiang WM, He Q, Tang KX (2016) Characterization of a trichome-specific promoter of the aldehyde dehydrogenase 1 (ALDH1) gene in Artemisia annua. Plant Cell Tissue Org Cult 126:469–480. CrossRefGoogle Scholar
  21. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. CrossRefPubMedGoogle Scholar
  22. Ma JW, Fu XQ, Zhang TT, Qian HM, Zhao JY (2019) Cloning and analyzing of chalcone isomerase gene (AaCHI) from Artemisia annua. Plant Cell Tissue Org Cult 137:45–54. CrossRefGoogle Scholar
  23. Moore M, Ullman C (2003) Recent developments in the engineering of zinc finger proteins. Brief Funct Genom 1:342–355. CrossRefGoogle Scholar
  24. Mukhopadhyay A, Vij S, Tyagi AK (2004) Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco. Proc Nat Acad Sci USA 101:6309–6314. CrossRefPubMedGoogle Scholar
  25. Olsson ME, Olofsson LM, Lindahl A-L, Lundgren A, Brodelius M, Brodelius PE (2009) Localization of enzymes of artemisinin biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry 70:1123–1128. CrossRefPubMedGoogle Scholar
  26. Paddon CJ, Westfall PJ, Pitera DJ et al (2013) High-level semi-synthetic production of the potent antimalarial artemisinin. Nature 496:528–532. CrossRefPubMedGoogle Scholar
  27. Peiffer M, Tooker JF, Luthe DS, Felton GW (2009) Plants on early alert: glandular trichomes as sensors for insect herbivores. New Phytol 184:644–656. CrossRefPubMedGoogle Scholar
  28. Ro DK, Paradise EM, Ouellet M et al (2006) Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature 440:940–943. CrossRefPubMedGoogle Scholar
  29. Schilmiller AL, Last RL, Pichersky E (2008) Harnessing plant trichome biochemistry for the production of useful compounds. Plant J 54:702–711. CrossRefPubMedGoogle Scholar
  30. Shen Q, Lu X, Yan TX et al (2016) The jasmonate-responsive AaMYC2 transcription factor positively regulates artemisinin biosynthesis in Artemisia annua. New Phytol 210:1269–1281. CrossRefPubMedGoogle Scholar
  31. Shen Q, Zhang LD, Liao ZH et al (2018) The genome of Artemisia annua provides insight into the evolution of asteraceae family and artemisinin biosynthesis. Mol Plant 11:776–788. CrossRefPubMedGoogle Scholar
  32. Shi P, Fu XQ, Shen Q et al (2018) The roles of AaMIXTA1 in regulating the initiation of glandular trichomes and cuticle biosynthesis in Artemisia annua. New Phytol 217:261–276. CrossRefPubMedGoogle Scholar
  33. Singh NP, Lai HC (2004) Artemisinin induces apoptosis in human cancer cells. Anticancer Res 24:2277–2280. CrossRefPubMedGoogle Scholar
  34. Solanke AU, Sharma MK, Tyagi AK, Sharma AK (2009) Characterization and phylogenetic analysis of environmental stress-responsive SAP gene family encoding A20/AN1 zinc finger proteins in tomato. Mol Genet Genom 282:153–164. CrossRefGoogle Scholar
  35. Sreedharan S, Shekhawat UKS, Ganapathi TR (2012) MusaSAP1, a A20/AN1 zinc finger gene from banana functions as a positive regulator in different stress responses. Plant Mol Biol 80:503–517. CrossRefPubMedGoogle Scholar
  36. Tang KX, Tang LO, Wang YL, Xin HL (2010) Composition of traditional Chinese medicine for reducing blood fat and preparation method thereof. The United States Patent US2012041058, 20120216Google Scholar
  37. Traw MB, Bergelson J (2003) Interactive effects of jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. Plant Physiol 133:1367–1375. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Tyagi H, Jha S, Sharma M, Giri J, Tyagi AK (2014) Rice SAPs are responsive to multiple biotic stresses and overexpression of OsSAP1, an A20/AN1 zinc-finger protein, enhances the basal resistance against pathogen infection in tobacco. Plant Sci 225:68–76. CrossRefPubMedGoogle Scholar
  39. Vij S, Tyagi AK (2006) Genome-wide analysis of the stress associated protein (SAP) gene family containing A20/AN1 zinc-finger(s) in rice and their phylogenetic relationship with Arabidopsis. Mol Genet Genom 276:565–575. CrossRefGoogle Scholar
  40. Wang G (2014) Recent progress in secondary metabolism of plant glandular trichomes. Plant Biotechnol 31:353–361. CrossRefGoogle Scholar
  41. Wang HZ, Han JL, Kanagarajan S, Lundgren A, Brodelius PE (2013) Trichome-specific expression of the amorpha-4,11-diene 12-hydroxylase (cyp71av1) gene, encoding a key enzyme of artemisinin biosynthesis in Artemisia annua, as reported by a promoter-GUS fusion. Plant Mol Biol 81:119–138. CrossRefPubMedGoogle Scholar
  42. Wasternack C, Hause B (2013) Jasmonates: biosynthesis, perception, signal transduction and action in plant stress response, growth and development. An update to the 2007 review in Annals of Botany. Ann Bot 111:1021–1058. CrossRefPubMedPubMedCentralGoogle Scholar
  43. White NJ (2008) Qinghaosu (Artemisinin): the price of success. Science 320:330–334. CrossRefPubMedGoogle Scholar
  44. WHO (2017) World malaria report 2017. Accessed 1 Apr 2017
  45. Xiao SH (2005) Development of antischistosomal drugs in China, with particular consideration to praziquantel and the artemisinins. Acta Trop 96:153–167. CrossRefPubMedGoogle Scholar
  46. Yan TX, Chen MH, Shen Q et al (2017) HOMEODOMAIN PROTEIN 1 is required for jasmonate-mediated glandular trichome initiation in Artemisia annua. New Phytol 213:1145–1155. CrossRefPubMedGoogle Scholar
  47. Yan TX, Li L, Xie LH et al (2018) A novel HD-ZIP IV/MIXTA complex promotes glandular trichome initiation and cuticle development in Artemisia annua. New Phytol 218:567–578. CrossRefPubMedGoogle Scholar
  48. Zhang L, Jing FY, Li FP et al (2009) Development of transgenic Artemisia annua (Chinese wormwood) plants with an enhanced content of artemisinin, an effective anti-malarial drug, by hairpin-RNA-mediated gene silencing. Biotechnol Appl Biochem 52:199–207. CrossRefPubMedGoogle Scholar
  49. Zhang FY, Fu XQ, Lv ZY et al (2015) A basic leucine zipper transcription factor, AabZIP1, connects abscisic acid signaling with artemisinin biosynthesis in Artemisia annua. Mol Plant 8:163–175. CrossRefPubMedGoogle Scholar
  50. Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Joint International Research Laboratory of Metabolic & Developmental Sciences, Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D CenterShanghai Jiao Tong UniversityShanghaiChina
  2. 2.College of PharmacyAnhui University of Chinese MedicineHefeiChina

Personalised recommendations