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Effects of hormones, carbon sources, and photoperiod on bulblet development in Lilium fargesii Franch during in vitro culture

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Abstract

Lilium fargesii Franch is a slow-growing species endemic to China with small-sized bulbs; however, the factors affecting bulblet enlargement remain unclear. Here, we show that 60 g·L− 1 sucrose and darkness are optimal conditions to increase the weight and expansion of bulblets. Both the weight gain and expansion rates of bulblets were further promoted by exogenous indole-3-butyric acid (IBA), the gibberellin (GA) inhibitor paclobutrazol (PBZ), and the jasmonic acid (JA) inhibitor diethyldithiocarbamic acid (DIECA), and were inhibited by exogenous benzylaminopurine (6-BAP) and methyl jasmonate (MeJA) under a 16-h light/8-h dark photoperiod. Exogenous application of GA3, 6-BAP, and MeJA, but not IBA, increased internal hormone contents; the inhibitors kynurenine (Kyn), PBZ, and DIECA decreased internal hormone levels, whereas the cytokinin inhibitor lovastatin had no effect. The weight gain and expansion rates of bulblets were promoted by high internal IBA and 6-BAP contents and low internal MeJA content. The roots number of bulblets was promoted by decreased IBA and increased GA3, and was inhibited by increased IBA and deceased 6-BAP. Endogenous GA3 or 6-BAP levels influenced the proliferation and total weight of daughter bulblets but not the formation of new leaves. Bulblets enlargement was positively associated with the proliferation and total weight of daughter bulblets and negatively associated with internal MeJA content. The number of new leaves was significantly negatively correlated with the weight gain, expansion, and roots of bulblets. These findings provide valuable insights for the in vitro preservation of L. fargesii to facilitate the protection and utilization of wild germplasm resources.

Key message

Variations in carbon source, photoperiod, hormone content affect bulblet development in L. fargesii Franch.

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Data availability

The data generated and analyzed during the current study are available from the corresponding author upon reasonable request. Data sharing does not apply to this article as no datasets were generated or analyzed during the current study.

Abbreviations

6-BAP:

Benzylaminopurine

DIECA:

Diethyldithiocarbamic acid

ER:

Expansion rate

GA:

Gibberellin

IBA:

Indole-3-butyric acid

Kyn:

Kynurenine

MeJA:

Methyl jasmonate

NAA:

1-Naphthalene acetic acid

NOL:

Number of leaves

NOR:

Number of roots

PBZ:

Paclobutrazol

PODB:

Proliferation rate of daughter bulbs

TWODB:

Total weight of daughter bulblets

WGR:

Weight gain rate

References

  • Askari N (2016) Aspects of bulblet growth of lily in vitro. PhD thesis. Wageningen University

  • Askari N, Visser RGF (2022) The role of scale explants in the growth of regenerating lily bulblets in vitro. Plant Cell Tiss Org 149(3):589–598. https://doi.org/10.1007/s11240-022-02328-y

    Article  CAS  Google Scholar 

  • Askari N, Visser R, Klerk G (2018) Growth of Lily bulblets in Vitro, a review. J Hortic Sci 5:133–143

    CAS  Google Scholar 

  • Chai Z, Fang JL, Huang CL, Huang R, Tan X, Chen BS, Yao W, Zhang MQ (2022) A novel transcription factor, ScAIL1, modulates plant defense responses by targeting DELLA and regulating gibberellin and jasmonic acid signaling in sugarcane. J Exp Bot 73(19):6727–6743. https://doi.org/10.1093/jxb/erac339

    Article  CAS  PubMed  Google Scholar 

  • Chen XJ, He JH, Tan GY, Liang J, Hou YX, Wang M, Wang BM (2019) Development of an enzyme-linked immunosorbent assay and a dipstick assay for the rapid analysis of resveratrol in grape berries Food Chem 291:132–138 https://doi.org/10.1016/j.foodchem.2019.04.023

  • Deswiniyanti NW, Lestari NKD (2020) In vitro propagation of bulbs using NAA and BAP plant growth regulator treatment. Kne Life Sci 32–45. https://doi.org/10.18502/kls.v5i2.6437

  • Du Y, Jia G (2010) The germplasm and utilization evaluation for wild flower resources of Liliaceae in Qinling Mountains of Shaanxi Province. J Fujian Coll Forestry 30(03):284–288

    Google Scholar 

  • Du S, Li D, Du X, Ding Q (2014) Study on tissue culture and rapid propagation technology of Lilium Fargesii. Mod Hortic 08(4):12–13

    Google Scholar 

  • Fu YY, Shu LL, Li HY, Zhang XM, Liu X, Ou ZY, Liang XM, Qi XY, Yang LP (2023) Establishment of highly efficient plant regeneration, callus transformation and analysis of botrytis cinerea-responsive PR promoters in Lilium brownii var. viridulum. Plants-Basel 12(10) https://doi.org/10.3390/plants12101992

  • Gao J, Wang Y, Huang H (2016) Embryogenic callus induction and plant regeneration of Lilium Fargesii. Bull Bot Res 36(1):6

    Google Scholar 

  • Gao SP, Zhu Y, Zhou LY, Fu XF, Lei T, Chen QB, Yu XF, Zhou YH, Li WJ, Hu J, Hu D, Si WH (2018) Sucrose signaling function on the formation and swelling of bulblets of Lilium sargentiae EH Wilson. Plant Cell Tiss Org 135(1):143–153. https://doi.org/10.1007/s11240-018-1451-4

    Article  CAS  Google Scholar 

  • He G, Yang P, Tang Y, Cao Y, Qi X, Xu L, Ming J (2020) Mechanism of exogenous cytokinins inducing bulbil formation in Lilium lancifolium in vitro. Plant Cell Rep 39(7):861–872. https://doi.org/10.1007/s00299-020-02535-x

    Article  CAS  PubMed  Google Scholar 

  • He GR, Yang PP, Cao YW, Tang YC, Wang L, Song M, Wang J, Xu LF, Ming J (2021) Cytokinin Type-B response regulators promote bulbil initiation in Lilium lancifolium. Int J Mol Sci 22(7). https://doi.org/10.3390/ijms22073320

  • Jamwal M, Sharma S, Jain RK, Dalal RPS (2016) Development of propagation protocol and expression studies on Asiatic lily hybrids (Lilium spp). Indian J Hortic 73(2):245–250. https://doi.org/10.5958/0974-0112.2016.00055.4

    Article  Google Scholar 

  • Jásik J, de Klerk GJ (2006) Effect of methyl jasmonate on morphology and dormancy development in lily bulblets regenerated. J Plant Growth Regul 25(1):45–51. https://doi.org/10.1007/s00344-005-0048-4

    Article  CAS  Google Scholar 

  • Jin GC, Qi JF, Zu HY, Liu ST, Gershenzon J, Lou YG, Baldwin IT, Li R (2023) Jasmonate-mediated gibberellin catabolism constrains growth during herbivore attack in rice. Plant Cell. https://doi.org/10.1093/plcell/koad191

    Article  PubMed  PubMed Central  Google Scholar 

  • Khalid A, Aftab F (2020) Effect of exogenous application of IAA and GA3 on growth, protein content, and antioxidant enzymes of Solanum tuberosum L. grown in vitro under salt stress. Vitro Cell Dev Biology - Plant 56(3):377–389. https://doi.org/10.1007/s11627-019-10047-x

    Article  CAS  Google Scholar 

  • Kumar S, Kashyap M, Sharma DR (2005) Regeneration and bulblet growth from lily bulbscale explants as affected by retardants, sucrose and irradiance. Biol Plant 49(4):629–632. https://doi.org/10.1007/s10535-005-0064-z

    Article  Google Scholar 

  • Kumar S, Awasthi V, Kanwar JK (2007) Influence of growth regulators and nitrogenous compounds on bulblet formation and growth in oriental lily. Hortic Sci 34(2):77–83. https://doi.org/10.17221/1853-Hortsci

    Article  CAS  Google Scholar 

  • Lapiz-Culqui YK, Meléndez-Mori JB, Mállap-Detquizán G, Tejada-Alvarado JJ, Vilca-Valqui NC, Huaman-Human E, Oliva M, Goñas M (2022) In vitro bulbification of five lily varieties: an effective method to produce quality seeds and flowers. Int J Agron 2022:8775989. https://doi.org/10.1155/2022/8775989

    Article  CAS  Google Scholar 

  • Leshem B, Lilien-Kipnis H, Steinitz B (1982) The effect of light and of explant orientation on the regeneration and subsequent growth of bulblets on Lilium longiflorum Thunb, bulb-scale sections cultured in vitro. Sci Hort 17(2):129–136. https://doi.org/10.1016/0304-4238(82)90005-X

    Article  Google Scholar 

  • Lestari kd, Deswiniyanti NW, Astarini IA, Arpiwi LM (2019) Callus and shoot induction of leaf culture Lilium longiflorum with NAA and BAP. Nusantara BioscienceDOI. https://doi.org/10.13057/nusbiosci/n110209

    Article  Google Scholar 

  • Li XY, Wang CX, Cheng JY, Zhang J, da Silva JAT, Liu XY, Duan X, Li TL, Sun HM (2014) Transcriptome analysis of carbohydrate metabolism during bulblet formation and development in Lilium davidii var. Unicolor. BMC Plant Biol 14. https://doi.org/10.1186/s12870-014-0358-4

  • Li T, Zhang YM, Wang D, Liu Y, Dirk LMA, Goodman J, Downie AB, Wang JM, Wang GY, Zhao TY (2017) Regulation of seed vigor by manipulation of raffinose family oligosaccharides in Maize and Arabidopsis. Mol Plant 10(12):1540–1555. https://doi.org/10.1016/j.molp.2017.10.014

    Article  CAS  PubMed  Google Scholar 

  • Li JR, Chai M, Zhu XP, Zhang XH, Li H, Wang D, Xing Q, Zhang JZ, Sun MY, Shi L (2019) Cloning and expression analysis of LoCCD8 during IAA-induced bulbils outgrowth in lily (oriental hybrid ‘Sorbonne’). J Plant Physiol 236:39–50. https://doi.org/10.1016/j.jplph.2019.03.002

    Article  CAS  PubMed  Google Scholar 

  • Li KH, Ren HH, Zhao W, Zhao XQ, Gan CM (2023) Factors affecting bulblet multiplication in bulbous plants. Sci Hort 312. https://doi.org/10.1016/j.scienta.2023.111837

  • Lian QL, Xin HB, Li XX, Zhong XH, Yin YL, Yi MF (2013) Isolation, characterization and expression analysis of the genes- and encoding the key enzymes involved in jasmonic acid biosynthesis in Gladiolus hybridus. Sci Hort 154:88–95. https://doi.org/10.1016/j.scienta.2013.02.004

    Article  CAS  Google Scholar 

  • Lv C, Wang Y, Wang L (2020) Study on the influencing factors of bud proliferation and bulblet swelling of Lilium fargesii in vitro. J Southwest Forestry Univ 40(1):38–45

    Google Scholar 

  • Murashige F, Skoog F (1962) A revised medium for rapid growth and bioassys with tobacco tissue cultures. Physiol Plant 15(3):473–497

    Article  CAS  Google Scholar 

  • Prokopiuk B, Cioć M, Maślanka M, Paw B (2018) Effects of light spectra and benzyl adenine on in vitro adventitious bulb and shoot formation of Lilium regale E. H. Wilson. Propag Ornam Plants 18:12–18

    Google Scholar 

  • Qin H, Deng ZA, Zhang CY, Wang YY, Wang J, Liu H, Zhang ZL, Huang RF, Zhang ZJ (2016) Rice GDP-mannose pyrophosphorylase OsVTC1-1 and OsVTC1-3 play different roles in ascorbic acid synthesis. Plant Mol Biol 90(3):317–327. https://doi.org/10.1007/s11103-015-0420-0

    Article  CAS  PubMed  Google Scholar 

  • Ren ZM, Zhang D, Jiao C, Li DQ, Wu Y, Wang XY, Gao C, Lin YF, Ruan YL, Xia YP (2022) Comparative transcriptome and metabolome analyses identified the mode of sucrose degradation as a metabolic marker for early vegetative propagation in bulbs of Lycoris. Plant J 112(1):115–134. https://doi.org/10.1111/tpj.15935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seng SS, Wu J, Sui JJ, Wu CY, Zhong XH, Liu C, Liu C, Gong BH, Zhang FQ, He JN, Yi MF (2016) ADP-glucose pyrophosphorylase gene plays a key role in the quality of corm and yield of cormels in gladiolus. Biochem Bioph Res Co 474(1):206–212. https://doi.org/10.1016/j.bbrc.2016.04.103

    Article  CAS  Google Scholar 

  • Seng SS, Wu J, Liang JH, Zhang FQ, Yang QY, He JN, Yi MF (2017) Silencing GhAGPL1 reduces the quality and quantity of corms and cormels in Gladiolus. J Am Soc Hortic Sci 142(2):119–U116. https://doi.org/10.21273/Jashs03944-16

    Article  CAS  Google Scholar 

  • Sevcíková H, Masková P, Tarkowská D, Masek T, Lipavská H (2017) Carbohydrates and gibberellins relationship in potato tuberization. J Plant Physiol 214:53–63. https://doi.org/10.1016/j.jplph2017.04.003

    Article  PubMed  Google Scholar 

  • Singh Anil AK, Kapri M, Sisodia A, Padhi M, Pal S (2018) Effect of GA3 and benzyladenine (BA) on growth and bulb production in Lily (Lilium longiflorum). Int J Curr Microbiol App Sci 7(6):1236–1240

  • Sohn HB, Lee HY, Seo JS, Jung C, Jeon JH, Kim JH, Lee YW, Lee JS, Cheong JJ, Do Choi Y (2011) Overexpression of jasmonic acid carboxyl methyltransferase increases tuber yield and size in transgenic potato. Plant Biotechnol Rep 5(1):27–34. https://doi.org/10.1007/s11816-010-0153-0

    Article  Google Scholar 

  • Song SL, Yan R, Wang CX, Wang JX, Sun HM (2020) Improvement of a genetic transformation system and preliminary study on the function of LpABCB21 and LpPILS7 based on somatic embryogenesis in Lilium pumilum DC. Fisch. International Journal of Molecular Sciences 21(18) https://doi.org/10.3390/ijms21186784

  • Su J, Jiao TT, Liu X, Zhu LC, Ma BQ, Ma FW, Li MJ (2023) Calcyclin-binding protein-promoted degradation of MdFRUCTOKINASE2 regulates sugar homeostasis in apple. Plant Physiol 191(2):1052–1065. https://doi.org/10.1093/plphys/kiac549

    Article  CAS  PubMed  Google Scholar 

  • Sun M, Shi JF, Zhang QX (2011) Effects of sucrose concentration on the bulblet formation and growth of three lilies in vitro. Adv Mater Res 343–344:1276–1280

    Article  Google Scholar 

  • Sun Q, Zhang B, Yang CL, Wang WL, Xiang L, Wang YP, Chan ZL (2022) Jasmonic acid biosynthetic genes TgLOX4 and TgLOX5 are involved in daughter bulb development in tulip (Tulipa gesneriana). Hortic Res-England 9. https://doi.org/10.1093/hr/uhac006

  • Tang N, Ju XT, Hu YF, Jia RL, Tang DC (2020) Effects of temperature and plant growth regulators on the scale propagation of Lilium davidii var. Unicolor. HortScience 55(6):870–875. https://doi.org/10.21273/Hortsci14916-20

    Article  CAS  Google Scholar 

  • Wu Y, Ren ZM, Gao C, Sun MY, Li SQ, Min RH, Wu J, Li DQ, Wang XY, Wei YP, Xia YP (2021) Change in sucrose cleavage pattern and papid starch accumulation govern Lily shoot-to-bulblet transition. Front Plant Sci 11. https://doi.org/10.3389/fpls.2020.564713

  • Xu JX, Li QZ, Yang LY, Li X, Wang Z, Zhang YC (2020) Changes in carbohydrate metabolism and endogenous hormone regulation during bulblet initiation and development in Lycoris. BMC Plant Biol 20(1). https://doi.org/10.1186/s12870-020-02394-4

  • Yadav R, Yadav N, Pal M, Goutam U (2013) Multiple shoot proliferation, bulblet induction and evaluation of genetic stability in Asiatic hybrid lily (Lilium Sp). Indian J Plant Physiol 18(4):354–359. https://doi.org/10.1007/s40502-014-0060-4

    Article  Google Scholar 

  • Yadav R, Yadav N, Pal M, Goutam U (2014) Multiple shoot proliferation, bulblet induction and evaluation of genetic stability in Asiatic hybrid lily (Lilium Sp). Indian J Plant Physiol 18. https://doi.org/10.1007/s40502-014-0060-4

  • Yang ZJ, Hao H, Zhou HJ, Liu QL (2019) Seed propagation and micropropagation of Lilium Fargesii. Acta Hort. https://doi.org/10.17660/ACTAHORTIC.2019.1237.31

    Article  Google Scholar 

  • Yin WZ, Yang HX, Wang YT, Feng P, Deng Y, Zhang LS, He GH, Wang N (2022) Oryza sativa PECTIN DEFECTIVE TAPETUM1 affects anther development through a pectin-mediated signaling pathway in rice. Plant Physiol 189(3):1570–1586. https://doi.org/10.1093/plphys/kiac172

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zeng Z, Lyu T, Jia X, Chen Y, Lyu YM (2022) Expression patterns of sugar transporter genes in the allocation of assimilates and abiotic stress in Lily. Int J Mol Sci 23(8). https://doi.org/10.3390/ijms23084319

  • Zhang Y, Zhang Q, Xue X (2010) The effects of the photoperiods on the bulblet formation and sugar metabolism change of wild Lilium lancifolium in vitro. Acta Horticulturae Sinica 37(6):957–962

    CAS  Google Scholar 

  • Zhang J, Gai MZ, Li XY, Li TL, Sun HM (2016) Somatic embryogenesis and direct as well as indirect organogenesis in Lilium pumilum DC. Fisch., an endangered ornamental and medicinal plant. Biosci Biotech Bioch 80(10):1898–1906. https://doi.org/10.1080/09168451.2016.1194178

    Article  CAS  Google Scholar 

  • Zhang BC, Gao YH, Zhang LJ, Zhou YH (2021) The plant cell wall: biosynthesis, construction, and functions. J Integr Plant Biol 63(1):251–272. https://doi.org/10.1111/jipb.13055

    Article  PubMed  Google Scholar 

  • Zhao J, Li G, Yi GX, Wang BM, Deng AX, Nan TG, Li ZH, Li QX (2006) Comparison between conventional indirect competitive enzyme-linked immunosorbent assay (icELISA) and simplified icELISA for small molecules. Anal Chim Acta 571(1):79–85. https://doi.org/10.1016/j.aca.2006.04.060

    Article  CAS  PubMed  Google Scholar 

  • Zheng RR, Wu Y, Xia YP (2012) Chlorocholine chloride and paclobutrazol treatments promote carbohydrate accumulation in bulbs of oriental hybrids ‘Sorbonne’. J Zhejiang Univ-Sc B 13(2):136–144. https://doi.org/10.1631/jzus.B1000425

    Article  CAS  Google Scholar 

  • Zhou L, Gao S, Chen F (2016) Action mechanism of sucrose and photoperiod on bulblet enlargement of lilium sargentiae. J Zhejiang Univ (Agric &Life Sci) 42(4):435–441

    Article  Google Scholar 

  • Zhu XP, Chai M, Li Y, Sun MY, Zhang JZ, Sun GF, Jiang CD, Shi L (2016) Global transcriptome profiling analysis of inhibitory effects of paclobutrazol on leaf growth in Lily (Longiflorum-Asiatic Hybrid). Front Plant Sci 7. https://doi.org/10.3389/fpls.2016.00491

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Acknowledgements

We would like to sincerely thank Prof. Cui Guangfen, Flower Research Institute, Yunnan Academy of Agricultural Sciences, for helping cultivate plantlets of Lilium fargesii Franch. We would like to also thank Prof. Wang Baomin, College of Agriculture, China Agricultural University, for providing the platform for internal hormone determination.

Funding

The research and demonstration on artificial domestication, breeding and cultivation technology for the national key protected wild flowers, the Public Welfare Industry (Agriculture) Research project, Ministry of Agriculture, No. 201203071, 2012–2016.

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  1. College of Horticulture, China Agricultural University, Beijing, 100193, China

    Li Zhou, Junna He & Qinglin Liu

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  1. Li Zhou
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  2. Junna He
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  3. Qinglin Liu
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All authors participated in the design, data collection, and statistical analysis. After the general framework was conceived by Prof. Dr. Liu Qinglin and Dr. He Junna, Miss Zhou wrote the manuscript, Liu Qinglin and He Junna revised and edited the manuscript, and all the authors read and finalized the manuscript.

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Zhou, L., He, J. & Liu, Q. Effects of hormones, carbon sources, and photoperiod on bulblet development in Lilium fargesii Franch during in vitro culture. Plant Cell Tiss Organ Cult 157, 30 (2024). https://doi.org/10.1007/s11240-024-02755-z

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