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Plant Molecular Biology

, Volume 98, Issue 3, pp 261–274 | Cite as

Molecular role of cytokinin in bud activation and outgrowth in apple branching based on transcriptomic analysis

  • Guofang Li
  • Ming Tan
  • Fang Cheng
  • Xiaojie Liu
  • Siyan Qi
  • Hongfei Chen
  • Dong Zhang
  • Caiping Zhao
  • Mingyu Han
  • Juanjuan Ma
Article

Abstract

Key message

Axillary bud activation and outgrowth were dependent on local cytokinin, and that bud activation preceded the activation of cell cycle and cell growth genes in apple branching.

Abstract

Cytokinin is often applied to apple trees to produce more shoot branches in apple seedlings. The molecular response of apple to the application of cytokinin, and the relationship between bud activation and cell cycle in apple branching, however, are poorly understood. In this study, RNA sequencing was used to characterize differential expression genes in axillary buds of 1-year grafted “Fuji” apple at 4 and 96 h after cytokinin application. And comparative gene expression analyses were performed in buds of decapitated shoots and buds of the treatment of biosynthetic inhibitor of cytokinin (Lovastatin) on decapitated shoots. Results indicated that decapitation and cytokinin increased ZR content in buds and internodes at 4–8 h, and induced bud elongation at 96 h after treatment, relative to buds in shoots receiving the Lovastatin treatment. RNA-seq analysis indicated that differential expression genes in auxin and cytokinin signal transduction were significantly enriched at 4 h, and DNA replication was enriched at 96 h. Cytokinin-responsive type-A response regulator, auxin polar transport, and axillary meristem-related genes were up-regulated at 4 h in the cytokinin and decapitation treatments, while qRT-PCR analysis showed that cell cycle and cell growth genes were up-regulated after 8 h. Collectively, the data indicated that bud activation and outgrowth might be dependent on local cytokinin synthesis in axillary buds or stems, and that bud activation preceded the activation of cell cycle genes during the outgrowth of ABs in apple shoots.

Keywords

Cytokinin Axillary bud Bud activation Cell cycle Apple 

Notes

Acknowledgements

National Apple Industry Technology System of Agriculture Ministry of China (CARS-28), Yangling Subsidiary Center Project of National Apple Improvement Center (C000088), Chinese postdoctoral project (2015M582713), Innovation project of science and technology plan projects of Shaanxi province (2016TZC-N-11-6). CARS-28 and 2015M582713 were involved in sample collection and RNA-seq data. C000088 and 2016TZC-N-11-6 were involved qPCR, hormone and sugar measurement, and manuscript revision.

Author contributions

GFL, MT, DZ, CPZ, MYH and JJM participated in the experimental design and data analysis. GFL, MT, FC, XJL, SYQ, and HFC performed material sampling, field measurements and the measurement of laboratory data. GFL and MT participated in the analysis of RNA-sEq. GFL, MT, MYH and JJM participated in the paper writing and revision. Sadly, Prof. Mingyu Han (MYH) has been deceased in August 2018.

Compliance with ethical standards

Competing interests

All authors agree with the submission and declare that they have no competing interests.

Supplementary material

11103_2018_781_MOESM1_ESM.doc (4.2 mb)
Supplementary material 1 (DOC 4295 KB)

References

  1. Arite T, Iwata H, Ohshima K, Maekawa M, Nakajima M, Kojima M, Sakakibara H, Kyozuka J (2007) DWARF10, an RMS1/MAX4/DAD1 ortholog, controls lateral bud outgrowth in rice. Plant J 51:1019–1029CrossRefGoogle Scholar
  2. Balla J, Kalousek P, Reinohl V, Friml J, Prochazka S (2011) Competitive canalization of PIN-dependent auxin flow from axillary buds controls pea bud outgrowth. Plant J 65:571–577CrossRefGoogle Scholar
  3. Barbier F, Peron T, Lecerf M, Perez-Garcia MD, Barriere Q, Rolcik J, Boutet-Mercey S, Citerne S, Lemoine R, Porcheron B, Roman H, Leduc N, Le Gourrierec J, Bertheloot J, Sakr S (2015) Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida. J Exp Bot 66:2569–2582CrossRefGoogle Scholar
  4. Benjamini Y, Yekutieli D (2001) The control of the false discovery rate in multiple testing under dependency. Ann Stat 29:1165–1188CrossRefGoogle Scholar
  5. Bennett T, Sieberer T, Willett B, Booker J, Luschnig C, Leyser O (2006) The Arabidopsis MAX pathway controls shoot branching by regulating auxin transport. Curr Biol 16:553–563CrossRefGoogle Scholar
  6. Beveridge CA, Murfet IC, Kerhoas L, Sotta B, Miginiac E, Rameau C (1997) The shoot controls zeatin riboside export from pea roots. Evidence from the branching mutant rms4. Plant J 11:339–345CrossRefGoogle Scholar
  7. Booker J, Chatfield S, Leyser O (2003) Auxin acts in xylem-associated or medullary cells to mediate apical dominance. Plant Cell 15:495–507CrossRefGoogle Scholar
  8. Braun N, de Saint Germain A, Pillot JP, Boutet-Mercey S, Dalmais M, Antoniadi I, Li X, Maia-Grondard A, Le Signor C, Bouteiller N, Luo D, Bendahmane A, Turnbull C, Rameau C (2012) The pea TCP transcription factor PsBRC1 acts downstream of strigolactones to control shoot branching. Plant Physiol 158:225–238CrossRefGoogle Scholar
  9. Brown BT, Foster C, Phillips JN, Rattigan BM (1979) The indirect role of 2,4-D in the maintenance of apical dominance in decapitated sunflower seedlings (Helianthus annuus L.). Planta 146:475–480CrossRefGoogle Scholar
  10. Chatfield SP, Stirnberg P, Forde BG, Leyser O (2000) The hormonal regulation of axillary bud growth in Arabidopsis. Plant J 24:159–169CrossRefGoogle Scholar
  11. Cline MG (1996) Exogenous auxin effects on lateral bud outgrowth in decapitated shoots. Ann Bot-London 78:255–266CrossRefGoogle Scholar
  12. Cook NC, Bellstedt DU, Jacobs G (2001) Endogenous cytokinin distribution patterns at budburst in Granny Smith and Braeburn apple shoots in relation to bud growth. Sci Hortic 87:53–63CrossRefGoogle Scholar
  13. Cubas P, Lauter N, Doebley J, Coen E (1999) The TCP domain: a motif found in proteins regulating plant growth and development. Plant J 18:215–222CrossRefGoogle Scholar
  14. de Jong M, George G, Ongaro V, Williamson L, Willetts B, Ljung K, McCulloch H, Leyser O (2014) Auxin and strigolactone signaling are required for modulation of arabidopsis shoot branching by nitrogen supply. Plant Physiol 166:384–395CrossRefGoogle Scholar
  15. Dello Ioio R, Linhares FS, Scacchi E, Casamitjana-Martinez E, Heidstra R, Costantino P, Sabatini S (2007) Cytokinins determine Arabidopsis root-meristem size by controlling cell differentiation. Curr Biol 17:678–682CrossRefGoogle Scholar
  16. Devitt ML, Stafstrom JP (1995) Cell cycle regulation during growth-dormancy cycles in pea axillary buds. Plant Mol Biol 29:255–265CrossRefGoogle Scholar
  17. Dun EA, Ferguson BJ, Beveridge CA (2006) Apical dominance and shoot branching. Divergent opinions or divergent mechanisms? Plant Physiol 142:812–819CrossRefGoogle Scholar
  18. Faiss M, Zalubilova J, Strnad M, Schmulling T (1997) Conditional transgenic expression of the ipt gene indicates a function for cytokinins in paracrine signaling in whole tobacco plants. Plant J 12:401–415CrossRefGoogle Scholar
  19. Giulini A, Wang J, Jackson D (2004) Control of phyllotaxy by the cytokinin-inducible response regulator homologue ABPHYL1. Nature 430:1031–1034CrossRefGoogle Scholar
  20. Gonzalez-Grandio E, Pajoro A, Franco-Zorrilla JM, Tarancon C, Immink RGH, Cubas P (2017) Abscisic acid signaling is controlled by a BRANCHED1/HD-ZIP I cascade in Arabidopsis axillary buds. Proc Natl Acad Sci USA 114:E245–E254CrossRefGoogle Scholar
  21. Hartig K, Beck E (2005) Assessment of lovastatin application as tool in probing cytokinin-mediated cell cycle regulation. Physiol Plant 125:260–267CrossRefGoogle Scholar
  22. Jiang L, Liu X, Xiong G, Liu H, Chen F, Wang L, Meng X, Liu G, Yu H, Yuan Y, Yi W, Zhao L, Ma H, He Y, Wu Z, Melcher K, Qian Q, Xu HE, Wang Y, Li J (2013) DWARF 53 acts as a repressor of strigolactone signalling in rice. Nature 504:401–405CrossRefGoogle Scholar
  23. Kalousek P, Buchtova D, Balla J, Reinoehl V, Prochazka S (2010) Cytokinins and polar transport of auxin in axillary pea buds. Magazine Acta Universitatis Agriculturae et Silviculturae Mendeleianae Brunensis 58:79–88CrossRefGoogle Scholar
  24. Kiba T, Aoki K, Sakakibara H, Mizuno T (2004) Arabidopsis response regulator, ARR22, ectopic expression of which results in phenotypes similar to the wol cytokinin-receptor mutant. Plant Cell Physiol 45:1063–1077CrossRefGoogle Scholar
  25. Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL (2013) TopHat2: accurate alignment of transcriptomesin the presence of insertions, deletions and gene fusions. Genome Biol 14:R36CrossRefGoogle Scholar
  26. Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12:357–360CrossRefGoogle Scholar
  27. King R, Vanstaden J (1988) Differential responses of buds along the shoot of pisum-sativum to isopentenyladenine and zeatin application. Plant Physiol Biochem 26:253–259Google Scholar
  28. Lazar G, Goodman HM (2006) MAX1, a regulator of the flavonoid pathway, controls vegetative axillary bud outgrowth in Arabidopsis. Proc Natl Acad Sci USA 103:472–476CrossRefGoogle Scholar
  29. Leibfried A, To JPC, Busch W, Stehling S, Kehle A, Demar M, Kieber JJ, Lohmann JU (2005) WUSCHEL controls meristem function by direct regulation of cytokinin-inducible response regulators. Nature 438:1172–1175CrossRefGoogle Scholar
  30. Leyser O (2003) Regulation of shoot branching by auxin. Trends Plant Sci 8:541–545CrossRefGoogle Scholar
  31. Li X, Mo XR, Shou HX, Wu P (2006) Cytokinin-mediated cell cycling arrest of pericycle founder cells in lateral root initiation of Arabidopsis. Plant Cell Physiol 47:1112–1123CrossRefGoogle Scholar
  32. Li G, Ma J, Tan M, Mao J, An N, Sha G, Zhang D, Zhao C, Han M (2016) Transcriptome analysis reveals the effects of sugar metabolism and auxin and cytokinin signaling pathways on root growth and development of grafted apple. BMC Genomics 17:150CrossRefGoogle Scholar
  33. Lin H, Wang RX, Qian Q, Yan MX, Meng XB, Fu ZM, Yan CY, Jiang B, Su Z, Li JY, Wang YH (2009) DWARF27, an iron-containing protein required for the biosynthesis of strigolactones, regulates rice tiller bud outgrowth. Plant Cell 21:1512–1525CrossRefGoogle Scholar
  34. Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15:550CrossRefGoogle Scholar
  35. Mason MG, Li J, Mathews DE, Kieber JJ, Schaller GE (2004) Type-B response regulators display overlapping expression patterns in Arabidopsis. Plant Physiol 135:927–937CrossRefGoogle Scholar
  36. Mason MG, Ross JJ, Babst BA, Wienclaw BN, Beveridge CA (2014) Sugar demand, not auxin, is the initial regulator of apical dominance. Proc Natl Acad Sci USA 111:6092–6097CrossRefGoogle Scholar
  37. Matsumoto-Kitano M, Kusumoto T, Tarkowski P, Kinoshita-Tsujimura K, Vaclavikova K, Miyawaki K, Kakimoto T (2008) Cytokinins are central regulators of cambial activity. Proc Natl Acad Sci USA 105:20027–20031CrossRefGoogle Scholar
  38. McCarthy DJ, Chen YS, Smyth GK (2012) Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Res 40:4288–4297CrossRefGoogle Scholar
  39. Medford JI, Horgan R, El-Sawi Z, Klee HJ (1989) Alterations of endogenous cytokinins in transgenic plants using a chimeric isopentenyl transferase gene. Plant Cell 1:403–413CrossRefGoogle Scholar
  40. Minakuchi K, Kameoka H, Yasuno N, Umehara M, Luo L, Kobayashi K, Hanada A, Ueno K, Asami T, Yamaguchi S, Kyozuka J (2010) FINE CULM1 (FC1) works downstream of strigolactones to inhibit the outgrowth of axillary buds in rice. Plant Cell Physiol 51:1127–1135CrossRefGoogle Scholar
  41. Miyawaki K, Matsumoto-Kitano M, Kakimoto T (2004) Expression of cytokinin biosynthetic isopentenyltransferase genes in Arabidopsis: tissue specificity and regulation by auxin, cytokinin, and nitrate. Plant J 37:128–138CrossRefGoogle Scholar
  42. Mortazavi A, Williams BA, Mccue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-SEq. Nat Methods 5:621–628CrossRefGoogle Scholar
  43. Mueller D, Leyser O (2011) Auxin, cytokinin and the control of shoot branching. Ann Bot-London 107:1203–1212CrossRefGoogle Scholar
  44. Muller D, Waldie T, Miyawaki K, To JPC, Melnyk CW, Kieber JJ, Kakimoto T, Leyser O (2015) Cytokinin is required for escape but not release from auxin mediated apical dominance. Plant J 82:874–886CrossRefGoogle Scholar
  45. Nguyen TQ, Emery RJN (2017) Is ABA the earliest upstream inhibitor of apical dominance? J Exp Bot 68:881–884CrossRefGoogle Scholar
  46. Ni J, Gao CC, Chen MS, Pan BZ, Ye KQ, Xu ZF (2015) Gibberellin promotes shoot branching in the perennial woody plant Jatropha curcas. Plant Cell Physiol 56:1655–1666CrossRefGoogle Scholar
  47. Nordstrom A, Tarkowski P, Tarkowska D, Norbaek R, Astot C, Dolezal K, Sandberg G (2004) Auxin regulation of cytokinin biosynthesis in Arabidopsis thaliana: a factor of potential importance for auxin-cytokinin-regulated development. Proc Natl Acad Sci USA 101:8039–8044CrossRefGoogle Scholar
  48. Pertea M, Kim D, Pertea GM, Leek JT, Salzberg SL (2016) Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Nat Protoc 11:1650–1667CrossRefGoogle Scholar
  49. Robinson MD, McCarthy DJ, Smyth GK (2010) edgeR: a bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26:139–140CrossRefGoogle Scholar
  50. Rosa M, Hilal M, Gonzalez JA, Prado FE (2009) Low-temperature effect on enzyme activities involved in sucrose-starch partitioning in salt-stressed and salt-acclimated cotyledons of quinoa (Chenopodium quinoa Willd.) seedlings. Plant Physiol Biochem 47:300–307CrossRefGoogle Scholar
  51. Shimizu-Sato S, Tanaka M, Mori H (2009) Auxin-cytokinin interactions in the control of shoot branching. Plant Mol Biol 69:429–435CrossRefGoogle Scholar
  52. Stirnberg P, Furner IJ, Leyser HMO (2007) MAX2 participates in an SCF complex which acts locally at the node to suppress shoot branching. Plant J 50:80–94CrossRefGoogle Scholar
  53. Strzalka W, Ziemienowicz A (2011) Proliferating cell nuclear antigen (PCNA): a key factor in DNA replication and cell cycle regulation. Ann Bot 107:1127–1140CrossRefGoogle Scholar
  54. Takei K, Ueda N, Aoki K, Kuromori T, Hirayama T, Shinozaki K, Yamaya T, Sakakibara H (2004) AtIPT3 is a key determinant of nitrate-dependent cytokinin biosynthesis in Arabidopsis. Plant Cell Physiol 45:1053–1062CrossRefGoogle Scholar
  55. Tanaka M, Takei K, Kojima M, Sakakibara H, Mori H (2006) Auxin controls local cytokinin biosynthesis in the nodal stem in apical dominance. Plant J 45:1028–1036CrossRefGoogle Scholar
  56. Tatusov RL, Galperin MY, Natale DA, Koonin EV (2000) The COG database: a tool for genome-scale analysis of protein functions and evolution. Nucleic Acids Res 28:33–36CrossRefGoogle Scholar
  57. Wang Y, Sun S, Zhu W, Jia K, Yang H, Wang X (2013) Strigolactone/MAX2-induced degradation of brassinosteroid transcriptional effector BES1 regulates shoot branching. Dev Cell 27:681–688CrossRefGoogle Scholar
  58. Xie X, Wang G, Yang L, Cheng T, Gao J, Wu Y, Xia Q (2015) Cloning and characterization of a novel Nicotiana tabacum ABC transporter involved in shoot branching. Physiol Plant 153:299–306CrossRefGoogle Scholar
  59. Xing LB, Zhang D, Li YM, Shen YW, Zhao CP, Ma JJ, An N, Han MY (2015) Transcription profiles reveal sugar and hormone signaling pathways mediating flower induction in apple (Malus domestica Borkh.). Plant Cell Physiol 56:2052–2068CrossRefGoogle Scholar
  60. Yao C, Finlayson SA (2015) Abscisic acid is a general negative regulator of Arabidopsis axillary bud growth. Plant Physiol 169:611–626CrossRefGoogle Scholar
  61. Yoneyama K, Kisugi T, Xie XN, Arakawa R, Ezawa T, Nomura T, Yoneyama K (2015) Shoot-derived signals other than auxin are involved in systemic regulation of strigolactone production in roots. Planta 241:687–698CrossRefGoogle Scholar
  62. Yoshida T, Mogami J, Yamaguchi-Shinozaki K (2014) ABA-dependent and ABA-independent signaling in response to osmotic stress in plants. Curr Opin Plant Biol 21:133–139CrossRefGoogle Scholar
  63. Zhao Z, Andersen SU, Ljung K, Dolezal K, Miotk A, Schultheiss SJ, Lohmann JU (2010) Hormonal control of the shoot stem-cell niche. Nature 465:1089–1092CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Guofang Li
    • 1
  • Ming Tan
    • 1
  • Fang Cheng
    • 1
  • Xiaojie Liu
    • 1
  • Siyan Qi
    • 1
  • Hongfei Chen
    • 1
  • Dong Zhang
    • 1
  • Caiping Zhao
    • 1
  • Mingyu Han
    • 1
  • Juanjuan Ma
    • 1
  1. 1.Department of Horticulture CollegeNorthwest Agriculture & Forestry UniversityYanglingChina

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