Journal of Plant Growth Regulation

, Volume 38, Issue 2, pp 359–372 | Cite as

Differential Effects of Paclobutrazol on the Bulblet Growth of Oriental Lily Cultured In Vitro: Growth Behavior, Carbohydrate Metabolism, and Antioxidant Capacity

  • Yun Wu
  • MinYi Sun
  • JiaPing Zhang
  • Lin Zhang
  • ZiMing Ren
  • RuiHan Min
  • XiuYun Wang
  • YiPing XiaEmail author


Bulblet growth is crucial for global lily production, and the applications of plant growth regulators have proven effective but with poor understanding of mechanisms. The aims of this study were to identify the effects of paclobutrazol (PBZ) on tube lilies. Low PBZ concentrations (5 × 10−4 mM, LPBZ) stimulated bulblet biomass, whereas higher doses suppressed the growth of the leaves and roots. Soluble carbohydrate and starch contents increased significantly with increased PBZ dose. The activities of adenosine 5′-diphosphate glucose pyrophosphorylase (AGPase), granule-bound starch synthase (GBSS), and soluble starch synthase (SSS) increased dramatically in response to PBZ treatments at later growth stages (60 days after transplanting) when carbon starvation occurred. In contrast, GBSS activity was enhanced throughout the whole growth period, indicating that the starch increase was attributed mainly to amylose synthesis. Carbohydrates were utilized more efficiently following LPBZ, with a relative bulblet weight of approximately 77.07%, which potentially ensured the source-sink balance. Alternatively, more carbohydrates were stored in response to high PBZ concentration. Intriguingly, PBZ usage significantly increased the activities of ascorbate peroxidase, catalase and glutathione reductase at the early stages, implicating a possible role in the elimination of ROS to maintain homeostasis. Application of LPBZ resulted in the largest bulblet, which weighed 396 mg (2.5 times that of the control) and measured 10.70 mm in diameter. The detailed characterizations of the bulblet swelling mechanism using PBZ in vitro offer suggestions of future PBZ usage in other bulbous crops.


Organ formation Paclobutrazol Granule-bound starch synthase Dynamic changes Source-sink balance Reactive oxygen species 



Special acknowledgments are given to the editors and reviewers. This research was financially supported by the National Natural Science Foundation of China (No. 31772337) and the China Postdoctoral Science Foundation (No. 2017M612005).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

344_2018_9844_MOESM1_ESM.tiff (3.8 mb)
Supplementary material 1. Fig. S1 Growth of control and the paclobutrazol treatments of in vitro Lilium Oriental Hybrids ‘Sorbonne’. The in vitro shoots were transplanted to media containing basal MS plus 8 g/L agar and 70 g/L sucrose with different doses of PBZ: 0 mM (CON), 5×10-4 mM (LPBZ), and 5×10-2 mM (HPBZ). DAT, days after transplanting. Bulblets were taken at 60 DAT. White bar=1 cm. (TIFF 3840 KB)


  1. Baninasab B, Ghobadi C (2011) Influence of paclobutrazol and application methods on high-temperature stress injury in cucumber seedlings. J Plant Growth Regul 30:213–219CrossRefGoogle Scholar
  2. Basu S, Roychoudhury A, Saha PP, Sengupta DN (2009) Differential antioxidative responses of indica rice cultivars to drought stress. Plant Growth Regul 60(1):51–59CrossRefGoogle Scholar
  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  4. Carver ST, Arnold MA, Byrne DH, Armitage AR, Lineberger RD, King AR (2014) Growth and flowering responses of sea marigold to daminozide, paclobutrazol, or uniconazole applied as drenches or sprays. J Plant Growth Regul 33:626–631CrossRefGoogle Scholar
  5. Chen J, Hall DE, De Luca V (2005) Effects of the growth retardant paclobutrazol on large-scale micropropagation of daylily (Hemerocallis spp.). In Vitro Cell Dev Biol Plant 41:58–62CrossRefGoogle Scholar
  6. Christiaens A, Pauwels E, Gobin B, Van Labeke MC (2015) Flower differentiation of azalea depends on genotype and not on the use of plant growth regulators. Plant Growth Regul 75:245–252CrossRefGoogle Scholar
  7. Currey CJ, Lopez RG (2010) Paclobutrazol pre-plant bulb dips effectively control height of ‘Nellie White’ Easter lily. HortTechnology 20:357–360CrossRefGoogle Scholar
  8. Dewir YHD, Chakrabarty D, Ali MB, Hahn EJ, Paek KY (2006) Lipid peroxidation and antioxidant enzyme activities of Euphorbia milii hyperhydric shoots. Environ Exp Bot 58:1553–1562CrossRefGoogle Scholar
  9. Elmongy MS, Zhou H, Cao Y, Liu B, Xia YP (2018) The effect of humic acid on endogenous hormone levels and antioxidant enzyme activity during in vitro rooting of evergreen azalea. Sci Hortic 227(Supplement C):234–243Google Scholar
  10. Eum SJ, Park KI, Lee IJ, Choi YJ, Oh W, Kim KW (2011) Effects of foliar-sprayed diniconazole on contents of endogenous gibberellic acids and abscisic acid in Lilium davuricum. Korean J Hortic Sci 29:165–171Google Scholar
  11. Foyer CH, Noctor G (2009) Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxid Redox Sign 11:861–905CrossRefGoogle Scholar
  12. Francescangeli N, Marinangeli P, Curvetto N (2007) Paclobutrazol for height control of two Lilium LA hybrids grown in pots. Span J Agric Res 5:425–430CrossRefGoogle Scholar
  13. Goleniowski M, Longo OD, Forchetti SMD, Argüello JA (2001) Relationships between peroxidases and in vitro bulbification in garlic (Allium sativum L.). In Vitro Cell Dev Biol Plant 37:683–686CrossRefGoogle Scholar
  14. Gomathinayagam M, Jaleel CA, Lakshmanan GMA, Panneerselvam R (2007) Changes in carbohydrate metabolism by triazole growth regulators in cassava (Manihot esculenta Crantz); effects on tuber production and quality. CR Biol 330:644–655CrossRefGoogle Scholar
  15. Grospietsch M, Lipavská H, Opatrná J (2000) Effect of paclobutrazol on soluble sugars and starch content of de novo regenerating potato stem explants. Biol Plant 43:137–139CrossRefGoogle Scholar
  16. Hajihashemi S, Ehsanpour AA (2014) Antioxidant response of Stevia rebaudiana B. to polyethylene glycol and paclobutrazol treatments under in vitro culture. Appl Biochem Biotechnol 172:4038–4052CrossRefGoogle Scholar
  17. Harvey BMR, Crothers SH, Evans NE, Selby C (1991) The use of growth-retardants to improve microtuber formation by potato (Solanum tuberosum). Plant Cell Tiss Org 27:59–64CrossRefGoogle Scholar
  18. He XL, Shi LW, Yuan ZH, Xu Z, Zhang ZQ, Yi MF (2008) Effects of lipoxygenase on the corm formation and enlargement in Gladiolus hybridus. Sci Hortic 118:60–69CrossRefGoogle Scholar
  19. Ilan A, Ziv M, Halevy AH (1995) Propagation and corm development of Brodiaea in liquid cultures. Sci Hortic 63:101–112CrossRefGoogle Scholar
  20. Jiao J, Tsujita M, Murr D (1986) Effects of paclobutrazol and A-Rest on growth, flowering, leaf carbohydrate and leaf senescence in ‘Nellie White’ Easter lily (Lilium longiflorum Thunb.). Sci Hortic 30:135–141CrossRefGoogle Scholar
  21. Jiao J, Wang X, Tsujita MJ (1991) Antagonistic effects of uniconazole and GA4+7 on shoot elongation and flower development in ‘Nellie White’ Easter lily. Sci Hortic 46:323–331CrossRefGoogle Scholar
  22. Kamoutsis AP, Chronopoulou-Sereli AG, Paspatis EA (1999) Paclobutrazol affects growth and flower bud production in gardenia under different light regimes. HortScience 34:674–675CrossRefGoogle Scholar
  23. Karalija E, Paric A, Muratović E (2013) Biochemical status of in vitro regenerated Lilium bosniacum and Lilium cattaniae plantlets. Cent Eur J Biol 8:912–920Google Scholar
  24. Koch K (2004) Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Curr Opin Plant Biol 7:235–246CrossRefGoogle Scholar
  25. Kumar S, Kashyap M, Sharma DR (2005) In vitro regeneration and bulblet growth from lily bulbscale explants as affected by retardants, sucrose and irradiance. Biol Plant 49:629–632CrossRefGoogle Scholar
  26. Langens-Gerrits MM, De Klerk GJM (1992) Dry-matter partitioning between bulbs and leaves in plantlets of Lilium speciosum regenerated in vitro. Acta Bot Neerl 41(4):461–468CrossRefGoogle Scholar
  27. Langens-Gerrits MM, De Klerk GJM (1999) Micropropagation of flower bulbs. Plant cell culture protocols. Springer, New York, pp 141–147CrossRefGoogle Scholar
  28. Langens-Gerrits MM, De Klerk GJM, Croes A (2003) Phase change in lily bulblets regenerated in vitro. Physiol Plant 119:590–597CrossRefGoogle Scholar
  29. Li M, Li J, Liu W, Liu L, Lu J, Niu J, Liu X, Yang Q (2014) A protocol for in vitro production of microtubers in Chinese yam (Dioscorea opposita). Biosci Biotechnol Biochem 78:1005–1009CrossRefGoogle Scholar
  30. Lian ML, Chakrabarty D, Paek KY (2003a) Growth of Lilium oriental hybrid ‘Casablanca’ bulblet using bioreactor culture. Sci Hortic 97:41–48CrossRefGoogle Scholar
  31. Lian ML, Murthy HN, Paek KY (2003b) Photoautotrophic culture conditions and photosynthetic photon flux influence growth of Lilium bulblets in vitro. In Vitro Cell Dev Biol Plant 39:532–535CrossRefGoogle Scholar
  32. Lian QL, Xin HB, Li XX, Zhong XH, Yin YL, Yi MF (2013) Isolation, characterization and expression analysis of the genes—GhAOS, GhAOC and GhOPR3: Encoding the key enzymes involved in jasmonic acid biosynthesis in Gladiolus hybridus. Sci Hortic 154:88–95CrossRefGoogle Scholar
  33. Lim S, Seon JH, Paek KY, Han BH, Son SH (1998) Development of pilot scale process for mass production of Lilium bulblets in vitro. International Society for Horticultural Science (ISHS), Leuven, pp 237–242Google Scholar
  34. Lin KH, Pai FH, Hwang SY, Lo HF (2006) Pre-treating paclobutrazol enhanced chilling tolerance of sweetpotato. Plant Growth Regul 49:249–262CrossRefGoogle Scholar
  35. McAvoy RJ (1991) Response of Easter lily to preplant incorporation of uniconazole into the planting medium. HortScience 26:152–154CrossRefGoogle Scholar
  36. Mccall I, Whipker BE, Krug BA (2005) Flurprimidol is effective at controlling height of ‘Star Gazer’ oriental lily. HortTechnology 15:373–376Google Scholar
  37. McCready R, Guggolz J, Silviera V, Owens H (1950) Determination of starch and amylose in vegetables. Anal Chem 22:1156–1158CrossRefGoogle Scholar
  38. Mehouachi J, Tadeo FR, Zaragoza S, Primo-Millo E, Talon M (1996) Effects of gibberellic acid and paclobutrazol on growth and carbohydrate accumulation in shoots and roots of citrus rootstock seedlings. J Jpn Soc Water Environ 31:1662–1672Google Scholar
  39. Misra P, Pandey V, Kochhar S (2010) Study of antioxidant enzymes activity during organogenesis and in vitro propagation of Asiatic hybrid lily. J Plant Biochem Biot 19:119–122CrossRefGoogle Scholar
  40. Moreno D, Berli FJ, Piccoli PN, Bottini R (2011) Gibberellins and abscisic acid promote carbon allocation in roots and berries of grapevines. J Plant Growth Regul 30:220–228CrossRefGoogle Scholar
  41. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  42. Niimi Y, Saito I (1990) Production of bulbs of Lilium rubellum Baker. An attempt to improve in vitro growth of bulblets regenerated from cultured bulb scales. J Jpn Soc Hortic Sci 59:635–640CrossRefGoogle Scholar
  43. Niimi Y, Misaki Y, Nakano M (2000) Production of commercial bulbs of Lilium rubellum Baker: changes in carbohydrates in bulblets and sugars of liquid medium during their culture. Engei Gakkai Zasshi 69:161–165CrossRefGoogle Scholar
  44. Okuda H, Kihara T, Iwagaki I (1996) Effects of paclobutrazol application to soil at the beginning of maturation on sprouting, shoot growth, flowering and carbohydrate contents in roots and leaves of Satsuma mandarin. J Hortic Sci 71(5):785–789CrossRefGoogle Scholar
  45. Petrić M, Jevremović SB, Trifunović M, Tadić V, Milošević S, Subotić (2014) Activity of antioxidant enzymes during induction of morphogenesis of Fritillaria meleagris in bulb scale culture. Turk J Biol 38:328–338CrossRefGoogle Scholar
  46. Pobudkiewicz A, Treder J (2006) Effects of flurprimidol and daminozide on growth and flowering of oriental lily ‘Mona Lisa’. Sci Hortic 110:328–333CrossRefGoogle Scholar
  47. Podwyszyńska M (2012) The Mechanisms of in vitro storage organ formation in ornamental geophytes. Floric Ornam Biotechnol 6:9–23Google Scholar
  48. Rachana S, Samiksha S, Parul P, Rohit KM, Durgesh KT, Vijay PS, Devendra KC, Sheo MP (2016) Reactive oxygen species (ROS): beneficial companions of plants developmental processes. Front Plant Sci 7:1299Google Scholar
  49. Ranwala AP, Legnani G, Reitmeier M, Stewart BB, Miller WB (2002) Efficacy of plant growth retardants as preplant bulb dips for height control in LA and oriental hybrid lilies. HortTechnology 12:426–431CrossRefGoogle Scholar
  50. Ranwala NKD, Ranwala AP, Miller WB (2005) Paclobutrazol and uniconazole solutions maintain efficacy after multiple lily bulb dip events. HortTechnology 15:551–553CrossRefGoogle Scholar
  51. Roberts AV, Matthews D (1995) The preparation in-vitro of chrysanthemum for transplantation to soil. Plant Cell Tiss Org 40:191–193CrossRefGoogle Scholar
  52. Sharifi G, Ebrahimzadeh H (2010) Changes of antioxidant enzyme activities and isoenzyme profiles during in vitro shoot formation in saffron (Crocus sativus L.). Acta Biol Hung 61:73–89CrossRefGoogle Scholar
  53. Sonnewald SJF, Uwe S (2012) The mode of sucrose degradation in potato tubers determines the fate of assimilate utilization. Front Plant Sci 3:279–286Google Scholar
  54. Srivastav M, Kishor A, Dahuja A, Sharma RR (2010) Effect of paclobutrazol and salinity on ion leakage, proline content and activities of antioxidant enzymes in mango (Mangifera indica L.). Sci Hortic 125:785–788CrossRefGoogle Scholar
  55. Tetlow IJ, Morell MK, Michael JE (2004) Recent developments in understanding the regulation of starch metabolism in higher plants. J Exp Bot 55(406):2131–2145CrossRefGoogle Scholar
  56. Thakur R, Sood A, Nagar PK, Pandey S, Sobti R, Ahuja PS (2006) Regulation of growth of Lilium plantlets in liquid medium by application of paclobutrazol or ancymidol, for its amenability in a bioreactor system: growth parameters. Plant Cell Rep 25:382–391CrossRefGoogle Scholar
  57. Tuyl JM, Arens P, Ramanna MS, Shahin A, Khan N, Xie SL, Marasek-Ciolakowska A, Lim KB, Barba-Gonzalez RB (2011) Lilium. Wild crop relatives: genomic and breeding resources. Springer, Heidelberg, pp 161–183CrossRefGoogle Scholar
  58. Van Harsselaar JK, Lorenz J, Senning M, Sonnewald U, Sonnewald S (2017) Genome-wide analysis of starch metabolism genes in potato (Solanum tuberosum L.). BMC Genom 18:37CrossRefGoogle Scholar
  59. Wang RJ, Wang ZL, Liang TB, Zhang XD, Liu LL, Shi CY (2008) Effects of HA-K fertilizers on the absorption and utilization of potassium and storage root yield in sweet potato for table use. Plant Nutr Fertil Sci 14:520–526Google Scholar
  60. Wen ZZ, Lin Y, Liu YQ, Wang M, Wang YQ, Liu W (2013) Effects of paclobutrazol in vitro on transplanting efficiency and root tip development of Dendrobium nobile. Biol Plant 57:576–580CrossRefGoogle Scholar
  61. Wu Y, Li Y, Ma YD, Zhang L, Ren ZM, Xia YP (2016a) Hormone and antioxidant responses of Lilium Oriental hybrids ‘Sorbonne’ bulblets to humic acid treatments in vitro. J Hortic Sci Biotechnol 92:1–13Google Scholar
  62. Wu Y, Xia YP, Zhang JP, Du F, Zhang L, Ma YD, Zhou H (2016b) Low humic acids promote in vitro lily bulblet enlargement by enhancing roots growth and carbohydrate metabolism. J Zhejiang Univ Sci B 17:892–904CrossRefGoogle Scholar
  63. Xia XJ, Zhou YH, Ding J, Shi K, Asami T, Chen Z, Yu JQ (2011) Induction of systemic stress tolerance by brassinosteroid in Cucumis sativus. New Phytol 191:706–720CrossRefGoogle Scholar
  64. Xiao YM, She LF, Chang L, Ren ZM, Xia YP (2013) Effect of three plant growth regulators on the bulblets development of Lycoris radiata. J Nucl Agric Sci 27:1409–1415Google Scholar
  65. Xu G, Luo R, Yao Y (2013) Paclobutrazol improved the reproductive growth and the quality of seed oil of Jatropha Curcas. J Plant Growth Regul 32:875–883CrossRefGoogle Scholar
  66. Yan H, Yang L, Li Y (2011) Axillary shoot proliferation and tuberization of Dioscorea fordii Prain et Burk. Plant Cell Tiss Org 104:193–198CrossRefGoogle Scholar
  67. Zheng RR, Xia YP, Sun XJ (2011) Uniconazole promotes carbohydrate accumulation in bulbs of Lilium oriental hybrid ‘Sorbonne’. Indian J Hortic 68:139–141Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Horticulture, College of Agriculture & BiotechnologyZhejiang UniversityHangzhouChina

Personalised recommendations