Advertisement

Plant Cell Reports

, Volume 33, Issue 9, pp 1479–1492 | Cite as

Methyl jasmonate represses growth and affects cell cycle progression in cultured Taxus cells

  • Rohan A. Patil
  • Sangram K. Lenka
  • Jennifer Normanly
  • Elsbeth L. Walker
  • Susan C. Roberts
Original Paper

Abstract

Key message

Methyl jasmonate elicitation of Taxus cultures enhances paclitaxel accumulation, but represses growth by inhibition of cell cycle progression. Growth repression is evident both at the culture level and transcriptional level.

Abstract

Methyl jasmonate (MeJA) elicitation is an effective strategy to induce and enhance synthesis of the anticancer agent paclitaxel (Taxol®) in Taxus cell suspension cultures; however, concurrent decreases in growth are often observed, which is problematic for large-scale bioprocessing. Here, increased accumulation of paclitaxel in Taxus cuspidata suspension cultures with MeJA elicitation was accompanied by a concomitant decrease in cell growth, evident within the first 3 days post-elicitation. Both MeJA-elicited and mock-elicited cultures exhibited similar viability with no apoptosis up to day 16 and day 24 of the cell culture period, respectively, suggesting that growth repression is not attributable to cell death. Flow cytometric analyses demonstrated that MeJA perturbed cell cycle progression of asynchronously dividing Taxus cells. MeJA slowed down cell cycle progression, impaired the G1/S transition as observed by an increase in G0/G1 phase cells, and decreased the number of actively dividing cells. Through a combination of deep sequencing and gene expression analyses, the expression status of Taxus cell cycle-associated genes correlated with observations at the culture level. Results from this study provide valuable insight into the mechanisms governing MeJA perception and subsequent events leading to repression of Taxus cell growth.

Keywords

Cell cycle Next-generation sequencing Methyl jasmonate Paclitaxel Plant cell culture Taxus 

Notes

Acknowledgments

This work was funded by National Institute of Health (Grant No. GM070852). We acknowledge the National Science Foundation-sponsored Institute for Cellular Engineering IGERT Program (DGE-0654128) for facilities and financial support. We thank Dr. Donna Gibson of the USDA, Agricultural Research Service, for the Taxus cell cultures. We also thank the UMass Amherst Flow Cytometry Facility and Prof. Shelly Peyton for the use of her Zeiss Axiovert 200 microscope.

Conflict of interest

The authors declare that they are no conflicts of interest.

Supplementary material

299_2014_1632_MOESM1_ESM.docx (203 kb)
Supplementary material 1 (DOCX 203 kb)
299_2014_1632_MOESM2_ESM.docx (23 kb)
Supplementary material 2 (DOCX 22 kb)
299_2014_1632_MOESM3_ESM.docx (22 kb)
Supplementary material 3 (DOCX 22 kb)
299_2014_1632_MOESM4_ESM.docx (22 kb)
Supplementary material 4 (DOCX 22 kb)

References

  1. Bonfill M, Exposito O, Moyano E, Cusido RM, Palazon J, Pinol MT (2006) Manipulation by culture mixing and elicitation of paclitaxel and baccatin III production in T. baccata suspension cultures. In Vitro Cell Dev Biol Plant 42:422–426Google Scholar
  2. Bonfill M, Bentebibel S, Moyano E, Palazón J, Cusidó R, Eibl R, Piñol M (2007) Paclitaxel and baccatin III production induced by methyl jasmonate in free and immobilized cells of Taxus baccata. Biol Plant 51:647–652Google Scholar
  3. Breyne P, Zabeau M (2001) Genome-wide expression analysis of plant cell cycle modulated genes. Curr Opin Plant Biol 4:136–142PubMedGoogle Scholar
  4. Bringi V, Kadkade P, Prince CL, and Roach B. (2007) Enhanced production of taxol and taxanes by cell cultures of Taxus species. US Patent-7264954 B1Google Scholar
  5. Castellano MM, del Pozo JC, Ramirez-Parra E, Brown S, Gutierrez C (2001) Expression and stability of Arabidopsis CDC6 are associated with endoreplication. Plant Cell 13:2671–2686PubMedCentralPubMedGoogle Scholar
  6. Cho K, Agrawal GK, Shibato J, Jung YH, Kim YK, Nahm BH et al (2007) Survey of differentially expressed proteins and genes in jasmonic acid treated rice seedling shoot and root at the proteomics and transcriptomics levels. J Proteome Res 6:3581–3603PubMedGoogle Scholar
  7. Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676PubMedGoogle Scholar
  8. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21Google Scholar
  9. Farmer EE, Ryan CA (1990) Interplant communication––airborne methyl jasmonate induces synthesis of proteinase-inhibitors in plant leaves. Proc Natl Acad Sci USA 87:7713–7716PubMedCentralPubMedGoogle Scholar
  10. Feiler HS, Desprez T, Santoni V, Kronenberger J, Caboche M, Traas J (1995) The higher-plant Arabidopsis thaliana encodes a functional Cdc48 homolog which is highly expressed in dividing and expanding cells. EMBO J 14:5626–5637PubMedCentralPubMedGoogle Scholar
  11. Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell-cycle in intact plant-tissues. Science 220:1049–1051PubMedGoogle Scholar
  12. Gaurav V, Kolewe ME, Roberts SC (2010) Flow cytometric methods to investigate culture heterogeneities for plant metabolic engineering. Methods Mol Biol 643:243–262PubMedGoogle Scholar
  13. Ge ZQ, Yuan YJ, Wang YD, Ma ZY, Hu ZD (2002) Ce4+-Induced apoptosis of T. cuspidata cells in suspension culture. J Rare Earths 20:139–144Google Scholar
  14. Goossens A, Hakkinen ST, Laakso I, Seppanen-Laakso T, Biondi S, De Sutter V, Lammertyn F, Nuutila AM, Soderlund H, Zabeau M, Inze D, Oksman-Caldentey KM (2003) A functional genomics approach toward the understanding of secondary metabolism in plant cells. Proc Natl Acad Sci USA 100:8595–8600PubMedCentralPubMedGoogle Scholar
  15. Gundlach H, Muller MJ, Kutchan TM, Zenk MH (1992) Jasmonic acid is a signal transducer in elicitor induced plant cell cultures. Proc Natl Acad Sci USA 89:2389–2393PubMedCentralPubMedGoogle Scholar
  16. Gutierrez C (2009) The Arabidopsis cell division cycle. Arabidopsis Book 7:e0120PubMedCentralPubMedGoogle Scholar
  17. Inze D, De Veylder L (2006) Cell cycle regulation in plant development. Annu Rev Genet 40:77–105PubMedGoogle Scholar
  18. Jennewein S, Wildung MR, Chau M, Walker K, Croteau R (2004) Random sequencing of an induced Taxus cell cDNA library for identification of clones involved in Taxol biosynthesis. Proc Natl Acad Sci USA 101:9149–9154PubMedCentralPubMedGoogle Scholar
  19. Kal AJ, van Zonneveld AJ, Benes V, van den Berg M, Koerkamp MG, Albermann K, Strack N, Ruijter JM, Richter A, Dujon B, Ansorge W, Tabak HF (1999) Dynamics of gene expression revealed by comparison of serial analysis of gene expression transcript profiles from yeast grown on two different carbon sources. Mol Biol Cell 10:1859–1872PubMedCentralPubMedGoogle Scholar
  20. Kawamura K, Murray JAH, Shinmyo A, Sekine M (2006) Cell cycle regulated D3-type cyclins form active complexes with plant-specific B-type cyclin-dependent kinase in vitro. Plant Mol Biol 61:311–327PubMedGoogle Scholar
  21. Ketchum REB, Gibson DM, Croteau RB, Shuler ML (1999) The kinetics of taxoid accumulation in cell suspension cultures of Taxus following elicitation with methyl jasmonate. Biotechnol Bioeng 62:97–105PubMedGoogle Scholar
  22. Kieran PM, MacLoughlin PF, Malone DM (1997) Plant cell suspension cultures: some engineering considerations. J Biotechnol 59:39–52PubMedGoogle Scholar
  23. Kim BJ, Gibson DM, Shuler ML (2004) Effect of subculture and elicitation on instability of Taxol production in Taxus sp suspension cultures. Biotechnol Progr 20:1666–1673Google Scholar
  24. Kim BJ, Gibson DM, Shuler ML (2005) Relationship of viability and apoptosis to taxol production in Taxus sp suspension cultures elicited with methyl jasmonate. Biotechnol Progr 21:700–707Google Scholar
  25. Kolewe ME, Henson MA, Roberts SC (2010) Characterization of aggregate size in Taxus suspension cell culture. Plant Cell Rep 29:485–494PubMedGoogle Scholar
  26. Kumagai-Sano F, Hayashi T, Sano T, Hasezawa S (2006) Cell cycle synchronization of tobacco BY-2 cells. Nat Protoc 1:2621–2627PubMedGoogle Scholar
  27. Laskaris G, Bounkhay M, Theodoridis G, van der Heijden R, Verpoorte R, Jaziri M (1999) Induction of geranylgeranyl diphosphate synthase activity and taxane accumulation in T. baccata cell cultures after elicitation by methyl jasmonate. Plant Sci 147:1–8Google Scholar
  28. Lenka SK, Boutaoui N, Paulose B, Vongpaseuth K, Normanly J, Roberts SC, Walker EL (2012) Identification and expression analysis of methyl jasmonate responsive ESTs in paclitaxel producing T. cuspidata suspension culture cells. BMC Genom 13:148Google Scholar
  29. Li J, Ou D, Zheng L, Gan N, Song L (2011) Applicability of the fluorescein diacetate assay for metabolic activity measurement of Microcystis aeruginosa (Chroococcales, Cyanobacteria). Phycol Res 59:200–207Google Scholar
  30. Li S, Zhang P, Zhang M, Fu CH, Zhao CF et al (2012) Transcriptional profile of Taxus chinensis cells in response to methyl jasmonate. BMC Genom 13:295Google Scholar
  31. Lijavetzky D, Almagro L, Belchi-Navarro S, Martinez-Zapater J, Bru R, Pedreno M (2008) Synergistic effect of methyljasmonate and cyclodextrin on stilbene biosynthesis pathway gene expression and resveratrol production in Monastrell grapevine cell cultures. BMC Res Notes 1:132PubMedCentralPubMedGoogle Scholar
  32. Logemann E, Wu SC, Schroder J, Schmelzer E, Somssich IE, Hahlbrock K (1995) Gene activation by UV light, fungal elicitor or fungal infection in Petroselinum crispum is correlated with repression of cell cycle-related genes. Plant J 8:865–876PubMedGoogle Scholar
  33. Mak YX, Doran PM (1993) Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells. Biotechnol Lett 15:1031–1034Google Scholar
  34. Meijer M, Murray JAH (2000) The role and regulation of D-type cyclins in the plant cell cycle. Plant Mol Biol 43:621–633PubMedGoogle Scholar
  35. Menges M, Hennig L, Gruissem W, Murray JAH (2002) Cell cycle-regulated gene expression in Arabidopsis. J Biol Chem 277:41987–42002PubMedGoogle Scholar
  36. Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat Methods 5:621–628PubMedGoogle Scholar
  37. Naill MC, Roberts SC (2005a) Cell cycle analysis of Taxus suspension cultures at the single cell level as an indicator of culture heterogeneity. Biotechnol Bioeng 90:491–500PubMedGoogle Scholar
  38. Naill MC, Roberts SC (2005b) Flow cytometric analysis of protein content in Taxus protoplasts and single cells as compared to aggregated suspension cultures. Plant Cell Rep 23:528–533PubMedGoogle Scholar
  39. Nims E, Dubois CP, Roberts SC, Walker EL (2006) Expression profiling of genes involved in paclitaxel biosynthesis for targeted metabolic engineering. Metab Eng 8:385–394PubMedGoogle Scholar
  40. Patil RA, Kolewe ME, Normanly J, Walker EL, Roberts SC (2012) Contribution of taxane biosynthetic pathway gene expression to observed variability in paclitaxel accumulation in Taxus suspension cultures. Biotechnol J 7:418–427PubMedCentralPubMedGoogle Scholar
  41. Patil RA, Kolewe ME, Roberts SC (2013) Cellular aggregation is a key parameter associated with long term variability in paclitaxel accumulation in Taxus suspension cultures. Plant Cell Tissue Organ Culture 112:303–310Google Scholar
  42. Pauwels L, Morreel K, Witte ED, Lammertyn F, Montagu MV, Boerjan W, Inzé D, Goossens A (2008) Mapping methyl jasmonate-mediated transcriptional reprogramming of metabolism and cell cycle progression in cultured Arabidopsis cells. Proc Natl Acad Sci USA 105:1380–1385PubMedCentralPubMedGoogle Scholar
  43. Pauwels L, Inzé D, Goossens A (2009) Jasmonate-inducible gene: what does it mean? Trends Plant Sci 14:87–91PubMedGoogle Scholar
  44. Piontkivska H, Rooney AP, Nei M (2002) Purifying selection and birth-and-death evolution in the histone H4 gene family. Mol Biol Evol 19:689–697PubMedGoogle Scholar
  45. Qiao JJ, Yuan YJ, Zhao H, Wu JC, Zeng AP (2003) Apoptotic cell death in suspension cultures of T. cuspidata co-treated with salicylic acid and hydrogen peroxide. Biotechnol Lett 25:387–390PubMedGoogle Scholar
  46. Reymond P, Farmer EE (1998) Jasmonate and salicylate as global signals for defense gene expression. Curr Opin Plant Biol 1:404–411PubMedGoogle Scholar
  47. Ruiz-May E, De-la-Peña C, Galaz-Ávalos RM, Lei Z, Watson BS, Sumner LW, Loyola-Vargas VM (2011) Methyl jasmonate induces ATP biosynthesis deficiency and accumulation of proteins related to secondary metabolism in Catharanthus roseus (L.) G. hairy roots. Plant Cell Physiol 52:1401–1421PubMedGoogle Scholar
  48. Ryerson DE, Heath MC (1996) Cleavage of nuclear DNA into oligonucleosomal fragments during cell death induced by fungal infection or by abiotic treatments. Plant Cell 8:393–402PubMedCentralPubMedGoogle Scholar
  49. Seo HS, Song JT, Cheong JJ, Lee YH, Lee YW, Hwang I, Lee JS, Choi YD (2001) Jasmonic acid carboxyl methyltransferase: a key enzyme for jasmonate-regulated plant responses. Proc Natl Acad Sci USA 98:4788–4793PubMedCentralPubMedGoogle Scholar
  50. Stevens R, Grelon M, Vezon D, Oh JS, Meyer P, Perennes C, Domenichini S, Bergounioux C (2004) A CDC45 homolog in Arabidopsis is essential for meiosis, as shown by RNA interference-induced gene silencing. Plant Cell 16:99–113PubMedCentralPubMedGoogle Scholar
  51. Sun G, Yang Y, Xie F, Wen J-F, Wu J, Wilson IW, Tang Q, Liu H, Qiu D (2013) Deep sequencing reveals transcriptome re-programming of Taxus × media cells to the elicitation with methyl jasmonate. PLoS One 8(4):1Google Scholar
  52. Swiatek A, Lenjou M, Van Bockstaele D, Inze D, Van Onckelen H (2002) Differential effect of jasmonic acid and abscisic acid on cell cycle progression in Tobacco BY-2 cells. Plant Physiol 128:201–211PubMedCentralPubMedGoogle Scholar
  53. Swiatek A, Van Dongen W, Esmans EL, Van Onckelen H (2004) Metabolic fate of jasmonates in tobacco bright yellow-2 cells. Plant Physiol 135:161–172PubMedCentralPubMedGoogle Scholar
  54. Takahashi N, Lammens T, Boudolf V, Maes S, Yoshizumi T, De Jaeger G, Witters E, Inze D, De Veylder L (2008) The DNA replication checkpoint aids survival of plants deficient in the novel replisome factor ETG1. EMBO J 27:1840–1851PubMedCentralPubMedGoogle Scholar
  55. Thanh NT, Murthy HN, Yu KW, Hahn EJ, Paek KY (2005) Methyl jasmonate elicitation enhanced synthesis of ginsenoside by cell suspension cultures of Panax ginseng in 5–l balloon type bubble bioreactors. Appl Microbiol Biotechnol 67:197–201PubMedGoogle Scholar
  56. Vandepoele K, Vlieghe K, Florquin K, Hennig L, Beemster GTS, Gruissem W, Van De Peer Y, Inze D, De Veylder L (2005) Genome-wide identification of potential plant E2F target genes. Plant Physiol 139:316–328PubMedCentralPubMedGoogle Scholar
  57. Vongpaseuth K, Roberts SC (2007) Advancements in the understanding of Paclitaxel metabolism in tissue culture. Curr Pharm Biotechnol 8:219–236PubMedGoogle Scholar
  58. Yang S, Lu SH, Yuan YJ (2008) Lipidomic analysis reveals differential defense responses of T. cuspidata cells to two elicitors, methyl jasmonate and cerium (Ce4+). Biochimica Et Biophysica Acta Mol 1781:123–134Google Scholar
  59. Yanpaisan W, King NJC, Doran PM (1998) Analysis of cell cycle activity and population dynamics in heterogeneous plant cell suspensions using flow cytometry. Biotechnol Bioeng 58:515–528PubMedGoogle Scholar
  60. Yazaki K, Takeda K, Tabata M (1997) Effects of methyl jasmonate on shikonin and dihydroechinofuran production in Lithospermum cell cultures. Plant Cell Physiol 38:776–782Google Scholar
  61. Yi H, Sardesai N, Fujinuma T, Chan CW, Veena, Gelvin SB (2006) Constitutive expression exposes functional redundancy between the Arabidopsis histone H2A gene HTA1 and other H2A gene family members. Plant Cell 18:1575–1589PubMedCentralPubMedGoogle Scholar
  62. Yuan Y-J, Li C, Hu Z-D, Wu J-C, Zeng A-P (2002) Fungal elicitor-induced cell apoptosis in suspension cultures of T. chinensis var. mairei for taxol production. Process Biochem 38:193–198Google Scholar
  63. Yukimune Y, Tabata H, Higashi Y, Hara Y (1996) Methyl jasmonate-induced overproduction of paclitaxel and baccatin III in Taxus cell suspension cultures. Nat Biotechnol 14:1129–1132PubMedGoogle Scholar
  64. Zhang Y, Turner JG (2008) Wound-induced endogenous jasmonates stunt plant growth by inhibiting mitosis. PLoS One 3:e3699PubMedCentralPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Rohan A. Patil
    • 1
  • Sangram K. Lenka
    • 2
  • Jennifer Normanly
    • 3
  • Elsbeth L. Walker
    • 2
  • Susan C. Roberts
    • 1
  1. 1.Department of Chemical Engineering, Institute for Cellular EngineeringUniversity of MassachusettsAmherstUSA
  2. 2.Department of BiologyUniversity of MassachusettsAmherstUSA
  3. 3.Department of Biochemistry and Molecular BiologyUniversity of MassachusettsAmherstUSA

Personalised recommendations