Plant Molecular Biology

, Volume 99, Issue 3, pp 205–217 | Cite as

The regulatory mechanism of chilling-induced dormancy transition from endo-dormancy to non-dormancy in Polygonatum kingianum Hemsl rhizome bud

  • Yue Wang
  • Xiaoqing Liu
  • He Su
  • Shikai Yin
  • Caixia Han
  • Dandan Hao
  • Xuehui DongEmail author


Key message

We identified three dormant stages of Polygonatum kingianum and changes that occurred during dormancy transition in the following aspects including cell wall and hormones, as well as interaction among them.


Polygonatum kingianum Hemsl (P. kingianum) is an important traditional Chinese medicine, but the mechanism of its rhizome bud dormancy has not yet been studied systematically. In this study, three dormancy phases were induced under controlled conditions, and changes occurring during the transition were examined, focusing on phytohormones and the cell wall. As revealed by HPLC–MS (High Performance Liquid Chromatography–Mass Spectrometry) analysis, the endo- to non-dormancy transition was association with a reduced abscisic acid (ABA)/gibberellin (GA3) ratio, a decreased level of auxin (IAA) and an increased level of trans-zeatin (tZR). Transmission electron microscopy showed that plasmodesmata (PDs) and the cell wall of the bud underwent significant changes between endo- and eco-dormancy. A total of 95,462 differentially expressed genes (DEGs) were identified based on transcriptomics, and clustering and principal component analysis confirmed the different physiological statuses of the three types of bud samples. Changes in the abundance of transcripts associated with IAA, cytokinins (CTKs), GA, ABA, brassinolide (BR), jasmonic acid (JA), ethylene, salicylic acid (SA), PDs and cell wall-loosening factors were analysed during the bud dormancy transition in P. kingianum. Furthermore, nitrilase 4 (NIT4) and tryptophan synthase alpha chain (TSA1), which are related to IAA synthesis, were identified as hub genes of the co-expression network, and strong interactions between hormones and cell wall-related factors were observed. This research will provide a good model for chilling-treated rhizome bud dormancy in P. kingianum and cultivation of this plant.


Rhizome bud dormancy Polygonatum kingianum RNA-Seq Plant hormone Cell wall 



I am very grateful to Ms. Liu for her help in the preparation of ultrathin sections and the RT-qPCR experiment. We also appreciate He Su for providing help about data analysis. I would like to thank Ms. Han, Mr. Yin and Ms. Hao for their assistance in the experiment. Most of all, I would like to thank professor Dong for his guidance on experimental consideration. We are particularly grateful to Liqing duojie, State Food and Drug Administration, Yunnan province for offering plantation base. We also express gratitude Yushi Zhang in College of Agronomy and Biotechnology, China Agricultural University for assistance of DEGs analysis. The funding for this work comes from Research on Seedling Breeding Technology of Paris Polyphylla granted by Chuansen Limited Company in Xishuangbanna.

Supplementary material

11103_2018_812_MOESM1_ESM.xlsx (12 kb)
Supplementary material 1 (XLSX 12 KB)
11103_2018_812_MOESM2_ESM.xlsx (10 kb)
Supplementary material 2 (XLSX 9 KB)
11103_2018_812_MOESM3_ESM.xlsx (32 kb)
Supplementary material 3 (XLSX 31 KB)
11103_2018_812_MOESM4_ESM.docx (1.2 mb)
Supplementary Figures (DOCX 1224 KB)
11103_2018_812_MOESM5_ESM.xlsx (98.4 mb)
DEG analysis of endo-, eco- and non-dormant rhizome buds (XLSX 100744 KB)
11103_2018_812_MOESM6_ESM.xlsx (691 kb)
Primary data used for principal component analysis and clustering analysis (XLSX 691 KB)
11103_2018_812_MOESM7_ESM.xlsx (26 kb)
Trend pathway analysis (XLSX 25 KB)
11103_2018_812_MOESM8_ESM.xlsx (73 kb)
Primary data used for gene co-expression analysis and clustering (XLSX 72 KB)
11103_2018_812_MOESM9_ESM.docx (44 kb)
The effect of GA3 on rhizome bud dormancy release (DOCX 43 KB)
11103_2018_812_MOESM10_ESM.docx (19 kb)
Supplementary Tables (DOCX 19 KB)
11103_2018_812_MOESM11_ESM.fa (418.6 mb)
Supplementary material 11 (FA 428689 KB)


  1. Anderson JV, Gesch RW, Jia Y, Chao WS, Horvath DP (2005) Seasonal shifts in dormancy status, carbohydrate metabolism, and related gene expression in crown buds of leafy spurge. Plant Cell Environ 28(12):1567–1578Google Scholar
  2. Bai S, Saito T, Sakamoto D, Ito A, Fujii H, Moriguchi T (2013) Transcriptome analysis of Japanese pear (Pyrus pyrifolia Nakai) flower buds transitioning through endodormancy. Plant Cell Physiol 54(7):1132Google Scholar
  3. Burchsmith TM, Zambryski PC (2010) Loss of increased size exclusion limit (ISE)1 or ISE2 increases the formation of secondary PD. Curr Biol 20(11):989–993Google Scholar
  4. Cantoro R, Crocco CD, Benecharnold RL, Rodríguez MV (2013) In vitro binding of Sorghum bicolor transcription factors ABI4 and ABI5 to a conserved region of a GA2-OXIDASE promoter: possible role of this interaction in the expression of seed dormancy. J Exp Bot 64(18):5721Google Scholar
  5. Chandler WH, Brown DS, Kimball MH, Philip GL, Tufts WP, Weldon AGP (1937) Chilling requirements for opening of buds on deciduous orchard trees and some other plants in California. California Agric Exp Sta Bull 611:1–63Google Scholar
  6. Chao WS, Dogramaci M, Horvath DP, Anderson JV, Foley ME (2017) Comparison of phytohormone levels and transcript profiles during seasonal dormancy transitions in underground adventitious buds of leafy spurge. Plant Mol Biol 94(3):281–302Google Scholar
  7. Chinese Pharmacopoeia Commission (2015) Pharmacopoeia of the people’s republic, 1st section. New York, pp 306–307Google Scholar
  8. Cho HT, Kende H (1997) Expression of expansin genes is correlated with growth in deepwater rice. Plant Cell 9(9):1661–1671Google Scholar
  9. Curaba J, Moritz T, Blervaque R, Parcy F, Raz V, Herzog M, Vachon G (2004) AtGA3ox2, a key gene responsible for bioactive gibberellin biosynthesis, is regulated during embryogenesis by LEAFY COTYLEDON2 and FUSCA3 in Arabidopsis. Plant Physiol 136(3):3660–3669Google Scholar
  10. Downes BP, Crowell DN (1998) Cytokinin regulates the expression of a soybean beta-expansin gene by a post-transcriptional mechanism. Plant Mol Biol 37(3):437–444Google Scholar
  11. Faust M, Erez A, Rowland LJ, Wang SY, Norman HA (1997) Bud dormancy in perennial fruit trees: physiological basis for dormancy induction, maintenance, and release. Hortscience 32(4):623–629Google Scholar
  12. Gazzarrini S, Tsuchiya Y, Lumba S, Okamoto M, Mccourt P (2004) The transcription factor FUSCA3 controls developmental timing in Arabidopsis through the hormones gibberellin and abscisic acid. Dev Cell 7(3):373–385Google Scholar
  13. Guak S, Neilsen D (2013) Chill unit models for predicting dormancy completion of floral buds in apple and sweet cherry. Hortic Environ Biotechnol 54(1):29–36Google Scholar
  14. Hao X, Yang Y, Yue C, Wang L, Horvath DP, Wang X (2017) Comprehensive transcriptome analyses reveal differential gene expression profiles of Camellia sinensis axillary buds at para-, endo-, ecodormancy, and bud flush stages. Front Plant Sci 8(481):553Google Scholar
  15. Horvath DP, Anderson JV, Chao WS, Foley ME (2003) Knowing when to grow: signals regulating bud dormancy. Trends Plant Sci 8(11):534–540Google Scholar
  16. Hutchison KW, Singer PB, Mcinnis S, Diazsala C, Greenwood MS (1999) Expansins are conserved in conifers and expressed in hypocotyls in response to exogenous auxin. Plant Physiol 120(3):827–832Google Scholar
  17. Jing W, Wu SS, Zhao LY, Jie L, Zhou LF, Zhu HH, Zhi C (2010) Genes related to the very early stage of ConA-induced fulminant hepatitis: a gene-chip-based study in a mouse model. BMC Genom 11(1):240Google Scholar
  18. Jones B, Gunneras SA, Petersson SV, Tarkowski P, Graham N, May S, Ljung K (2010) Cytokinin regulation of auxin synthesis in Arabidopsis involves a homeostatic feedback loop regulated via auxin and cytokinin signal transduction. Plant Cell 22(9):2956–2969Google Scholar
  19. Kalousek P, Buchtová D, Balla J, Reinöhl V, Procházka S (2014) Cytokinins and polartransport of auxin in axillary pea buds. Acta Univer Agric Silviculturae Mendelianae Brunensis 58(4):79–88Google Scholar
  20. Kauth PJ, Kane ME, Vendrame WA (2011) Chilling relieves corm dormancy in Calopogon tuberosus (Orchidaceae) from geographically distant populations. Environ Exp Bot 70(2–3):283–288Google Scholar
  21. Kim JH, Cho HT, Kende H (2000) α-Expansins in the semiaquatic ferns Marsilea quadrifolia and Regnellidium diphyllum: evolutionary aspects and physiological role in rachis elongation. Planta 212(1):85–92Google Scholar
  22. Kumar G, Gupta K, Pathania S, Swarnkar MK, Rattan UK, Singh G, Singh AK (2017) Chilling affects phytohormone and post-embryonic development pathways during bud break and fruit set in apple (Malus domestica Borkh.). Sci Rep 7:42593Google Scholar
  23. Lang GA, Martin GC, Darnell RL (1987) Endo-, para- and eco-dormancy: physiological terminology and classifcation for dormancy research. Hortscience 22:371–377Google Scholar
  24. Lattoo SK, Dhar AK, Jasrotia A (2001) Epicotyl seed dormancy and phenology of germination in Polygonatum cirrhifolium Royle. Curr Sci 81:1414–1417Google Scholar
  25. Mornya PMP, Cheng F (2013) Seasonal changes in endogenous hormone and sugar contents during bud dormancy in tree peony. J Appl Hortic 15:159–165Google Scholar
  26. Mornya PMP, Cheng FY, Li HY (2011) Chronological changes in plant hormone and sugar contents in cv. ao-shuang autumn flowering tree peony. Horticl Sci 38:104–112Google Scholar
  27. Mueller D, Leyser O (2011) Auxin, cytokinin and the control of shoot branching. Ann Bot 107:1203–1212Google Scholar
  28. Müssig C, Lisso J, Collgarcia D, Altmann T (2006) Molecular analysis of brassinosteroid action. Plant Biol 8:291–296Google Scholar
  29. Olszewski N, Gubler F (2002) Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell 14:S61Google Scholar
  30. Ongaro V, Leyser O (2008) Hormonal control of shoot branching. J Exp Bot 59:67Google Scholar
  31. Piotrowski MS, Schönfelder S, Weiler EW (2001) The Arabidopsis thaliana isogene NIT4 and its orthologs in tobacco encode beta-cyano-L-alanine hydratase/nitrilase. J Biol Chem 276:2616–2621Google Scholar
  32. Priti K, Tawhidur R, Divi UK (2010) Brassinosteroid-mediated stress tolerance in Arabidopsis shows interactions with abscisic acid, ethylene and salicylic acid pathways. BMC Plant Biol 10:151Google Scholar
  33. Rhie YH, Jung HH, Kim KS (2012) Chilling requirement for breaking dormancy and flowering in Paeonia lactiflora ‘Taebaek’. and ‘Mulsurae’. Hortic Environ Biotechnol 53:277–282Google Scholar
  34. Rinne PL, Welling A, Vahala J, Ripel L, Ruonala R, Kangasjã¤Rvi J, Van DSC (2011) Chilling of dormant buds hyperinduces FLOWERING LOCUS T and recruits GA-inducible 1,3-beta-glucanases to reopen signal conduits and release dormancy in Populus. Plant Cell 23:130–146Google Scholar
  35. Rodrigo MJ, Alquezar B, Zacarias L (2006) Cloning and characterization of two 9-cis-epoxycarotenoid dioxygenase genes, differentially regulated during fruit maturation and under stress conditions, from orange (Citrus sinensis L. Osbeck). J Exp Bot 57:633–643Google Scholar
  36. Takayama T, Toyomasu T, Yamane H, Murofushi N, Yajima H (2008) Identification of gibberellins and abscisic acid in bulbs of Lilium elegans thunb and their quantitative changes during cold treatment and the subsequent cultivation. Engei Gakkai Zasshi 62:189–196Google Scholar
  37. Tanimoto E (2005) Regulation of root growth by plant hormones—roles for auxin and gibberellin. Crit Rev Plant Sci 24:249–265Google Scholar
  38. Tylewicz S, Petterle A, Marttila S, Miskolczi P, Azeez A, Singh RK, Eklund DM (2018) Photoperiodic control of seasonal growth is mediated by ABA acting on cell-cell communication. Science. Google Scholar
  39. Van der Schoot C, Rinne PL (2011) Dormancy cycling at the shoot apical meristem: transitioning between self-organization and self-arrest. Plant Sci 180:120–131Google Scholar
  40. Winter N, Kollwig G, Zhang S, Kragler F (2007) MPB2C, a microtubule-associated protein, regulates non-cell-autonomy of the homeodomain protein KNOTTED1. Plant Cell 19:3001–3018Google Scholar
  41. Xu J, Xu Z, Zhu Y, Luo H, Qian J, Ji A, Song J (2014) Identification and evaluation of reference genes for qRT-PCR normalization in Ganoderma lucidum. Curr Microbiol 68:120–126Google Scholar
  42. Xu H, Cao D, Chen Y, Wei D, Wang Y, Stevenson RA, Lin J (2016) Gene expression and proteomic analysis of shoot apical meristem transition from dormancy to activation in Cunninghamia lanceolata (Lamb.). Hook Sci Rep 6:19938Google Scholar
  43. Yamazaki H, Nishijima T, Koshioka M, Miura H (2002) Gibberellins do not act against abscisic acid in the regulation of bulb dormancy of Allium wakegi Araki. Plant Growth Regul 36:223–229Google Scholar
  44. Yañez P, Ohno H, Ohkawa K (2005) Temperature effects on corm dormancy and growth of Zephyra elegans D.Don. Sci Hortic 105:127–138Google Scholar
  45. Yun NY, Rhie YH, Jung HH, Kim KS (2012) Chilling requirement for dormancy release of variegated Solomon’s seal. Hortic Environ Biotechnol 52:553–558Google Scholar
  46. Zhang JP (2015) Heat resistance evaluation and dormancy mechanism of the underground renewal bud of Paeonia lactiflora cultivated in Hangzhou city. Zhejiang UniversityGoogle Scholar
  47. Zhang R, Wang B, Ouyang J, Li JY, Wang YH (2008) Arabidopsis Indole Synthase,a homolog of Tryptophan Synthase Alpha,is an enzyme involved in the trp-independent indole-containing metabolite biosynthesis. Chinese Bull Bot 50:1070–1077Google Scholar
  48. Zhang Z, Zhuo X, Zhao K, Zheng T, Han Y, Yuan C, Zhang Q (2018) Transcriptome profiles reveal the crucial roles of hormone and sugar in the bud dormancy of Prunus mume. Sci Rep 8:5090Google Scholar
  49. Zhao YJ, Li LG (2011) Plant cell wal loosening factors. Plant Physiol J 47:925–935Google Scholar
  50. Zheng C, Halaly T, Acheampong AK, Takebayashi Y, Jikumaru Y, Kamiya Y, Or E (2015) Abscisic acid (ABA) regulates grape bud dormancy, and dormancy release stimuli may act through modification of ABA metabolism. J Exp Bot 66:1527–1542Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.College of Agronomy and BiotechnologyChina Agricultural UniversityBeijingChina

Personalised recommendations