Abstract
The combination of in vitro culture and simulated gravity under clinostat 2D conditions was first applied to strawberry (Fragaria × ananassa Duch.) leaf to evaluate in vitro morphogenesis, antioxidant enzyme activity and accumulation of endogenous hormones. The subsequent growth and runner formation of clinostat 2D generated-plantlets in the greenhouse as compared to those with plantlets in the in vitro culture condition (control) were also examined. The results showed that the leaf explants (0.5 cm × 0.5 cm) cultured under clinostat 2D gave 1-week shoot regeneration and the growth was higher than that under control condition after 3, 4 and 6 weeks of culture. In addition, in shoots generated from explants under clinostat 2D condition antioxidant enzyme activities affected results such as increasing ascorbate peroxidase (APX), catalase (CAT), 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and reducing phenolic content compared with control conditions after 6 weeks of culture. The clinostat 2D derived-shoots showed higher levels of GA3, ABA, and KIN; meanwhile, the content of IAA, ZEA, SA and MEL showed opposite results compared to the control condition. In this study, the ratios between endogenous hormone groups were also different between the two conditions. The ratios of GA3/IAA and GA3/CYT (ZEA, KIN and 2iP) of clinostat 2D derived-shoots were 12 and 1.5 times higher, respectively, compared to those in the control condition. In contrast, the ratios of IAA/CYT and IAA/SA showed the opposite results (two times lower) after 6 weeks of culture. The growth of shoots and plantlets derived from both conditions showed no abnormalities. In addition, the 2D clinostat generated-plantlets resulted in acclimatization and runner formation that was better than in the control. The results of the study offer a promising new direction for combining in vitro culture and clinostat 2D conditions for plant breeding.
Key message
In vitro culture and clinostat 2D conditions were applied to study leaf morphogenesis and 3 clinostat 2D-derived subsequent growth strawberry plantlets 4 Morphogenesis of in vitro strawberry leaf cultured under clinostat 2D were compared to 5 those under in vitro culture 6 Subsequent growth and runner formation of clinostat 2D-derived plantlets grown in the 7 greenhouse.
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Abbreviations
- 2iP:
-
N6-isopentenyladenine
- ABA :
-
Abscisic acid
- APX :
-
Ascorbate peroxidase
- CAT:
-
Catalase
- CYT:
-
Cytokinines
- DPPH:
-
2,2-Diphenyl-1-picrylhydrazyl
- GA3 :
-
Gibberellin A3
- HPLC:
-
High-performance liquid chromatography
- IAA:
-
Indole-3-acetic acid
- KIN:
-
Kinetin
- MEL:
-
N-acetyl-5-methoxytryptamine/melatonin
- MT:
-
6 -(3-hydroxybenzylamino) purine/ meta-topolin
- SA :
-
Salicylic acid
- ZEA :
-
Trans-zeatin.
References
Boucheron-Dubuisson E, Manzano AI, Le Disquet I, Matía I, Sáez-Vasquez J, Van Loon JJ, Medina FJ (2016) Functional alterations of root meristematic cells of Arabidopsis thaliana induced by a simulated microgravity environment. J Plant Physiol 207:30–41. https://doi.org/10.1016/j.jplph.2016.09.011
Cai W, Chen H, Jin J, Xu P, Bi T, Xie Q, Hu J (2019) Plant adaptation to microgravity environment and growth of plant cells in altered gravity conditions. In: Duan E, Long M (eds) Life science in space: experiments on board the SJ-10 recoverable satellite. Springer, Mainland China, pp 131–166
Cavallaro V, Pellegrino A, Muleo R, Forgione I (2022) Light and plant growth regulators on in vitro proliferation. Plants 11(7):844. https://doi.org/10.3390/plants11070844
Danova K, Motyka V, Todorova M, Trendafilova A, Krumova S, Dobrev P, Evstatieva L (2018) Effect of cytokinin and auxin treatments on morphogenesis, terpenoid biosynthesis, photosystem structural organization, and endogenous isoprenoid cytokinin profile in Artemisia alba Turra in vitro. J Plant Growth Regul 37(2):403–418. https://doi.org/10.1007/s00344-017-9738-y
Desmarchelier C, Bermudez MJN, Coussio J, Ciccia G, Boveris A (1997) Antioxidant and prooxidant activities in aqueous extract of Argentine plants. Int J Pharmacogn 35(2):116–120. https://doi.org/10.1076/phbi.35.2.116.13282
Dinani ET, Shukla MR, Turi CE, Sullivan JA, Saxena PK (2018) Thidiazuron: modulator of morphogenesis in vitro. In: Ahmad N, Faisal M (eds) Thidiazuron: from urea derivative to plant growth regulator. Springer, Singapore, pp 1–36
Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11(1):1–42. https://doi.org/10.2307/3001478
Ezura H, Harberd NP (1995) Endogenous gibberellin levels influence in vitro shoot regeneration in Arabidopsis thaliana (L.) Heynh. Planta 197(2):301–305. https://doi.org/10.1007/BF00202651
Finkelstein R (2013) Abscisic acid synthesis and response. Arabidopsis Book 11:1–36. https://doi.org/10.1199/tab.0166
Frolov A, Didio A, Ihling C, Chantzeva V, Grishina T, Hoehenwarter W, Medvedev S (2018) The effect of simulated microgravity on the Brassica napus seedling proteome. Funct Plant Biol 45(4):440–452. https://doi.org/10.1071/FP16378
Goth L (1991) A simple method for determination of serum catalase activity and revision of reference range. Clin Chim Acta 196(2–3):143–151. https://doi.org/10.1016/0009-8981(91)90067-m
Halimeh H (2022) Antioxidant metabolism and oxidative damage in Anthemis gilanica cell line under fast clinorotation. Plant Cell Tiss Org Cult 150(1):710–719. https://doi.org/10.1007/s11240-022-02324-2
Hoson T, Soga K, Mori R, Saiki M, Wakabayashi K, Kamisaka S, Shimazu T (1999) Morphogenesis of rice and Arabidopsis seedlings in space. J Plant Res 112(4):477–486. https://doi.org/10.1007/pl00013903
Huang P, Ju HW, Min JH, Zhang X, Chung JS, Cheong HS, Kim CS (2012) Molecular and physiological characterization of the Arabidopsis thaliana oxidation-related zinc finger 2, a plasma membrane protein involved in ABA and salt stress response through the ABI2-mediated signaling pathway. Plant Cell Physiol 53(1):193–203. https://doi.org/10.1093/pcp/pcr162
Jiménez VM, Guevara E, Herrera J, Bangerth F (2001) Endogenous hormone levels in habituated nucellar Citrus callus during the initial stages of regeneration. Plant Cell Rep 20(1):92–100. https://doi.org/10.1007/s002990000280
Khai HD, Bien LT, Vinh NQ, Dung DM, Nghiep ND, Mai NTN, Tung HT, Luan VQ, Cuong DM, Nhut DT (2021) Alterations in endogenous hormone levels and energy metabolism promoted the induction, differentiation and maturation of Begonia somatic embryos under clinorotation. Plant Sci 312:111045. https://doi.org/10.1016/j.plantsci.2021.111045
Lionheart G, Vandenbrink JP, Hoeksema JD, Kiss JZ (2018) The impact of simulated microgravity on the growth of different genotypes of the model legume plant Medicago truncatula. Microg Sci Technol 30:491–502. https://doi.org/10.1007/s12217-018-9619-4
Litwińczuk W, Wadas-Boroń M (2009) Development of highbush blueberry (Vaccinium corymbosum Hort. Non L.) in vitro shoot cultures under the influence of melatonin. Acta Sci Pol Hort Cultus 8(3):3–12
Manzano AI, Matía I, González-Camacho F, Carnero-Díaz E, van Loon JJ, Dijkstra C, Medina FJ (2009) Germination of Arabidopsis seed in space and in simulated microgravity: alterations in root cell growth and proliferation. Microg Sci Technol 21(4):293–297. https://doi.org/10.1007/s12217-008-9099-z
Mercier H, Souza BM, Hamasaki RM, Kraus JE, Sotta B (2003) Endogenous auxin and cytokinin contents associated with shoot formation in leaves of pineapple cultured in vitro. Braz J Plant Physiol 15(2):107–112. https://doi.org/10.1590/S1677-04202003000200006
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with Tobacco tissue cultures. Plant Physiol 1(3):473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Murata Y, Mori IC, Munemasa S (2015) Diverse stomatal signaling and the signal integration mechanism. Annu Rev Plant Biol 66:369–392. https://doi.org/10.1146/annurev-arplant-043014-114707
Nakajima S, Ogawa Y, Suzuki T, Kondo N (2019) Enhanced antioxidant activity in mung bean seedlings grown under slow clinorotation. Microg Sci Technol 31:395–401. https://doi.org/10.1007/s12217-019-9699-9
Nakajima S, Nagata M, Ikehata A (2021) Mechanism for enhancing the growth of mung bean seedlings under simulated microgravity. NPJ Microg 7(1):26. https://doi.org/10.1038/s41526-021-00156-6
Nakamura M, Nishimura T, Morita MT (2019) Gravity sensing and signal conversion in plant gravitropism. J Exp Bot 70(14):3495–3506. https://doi.org/10.1093/jxb/erz158
Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22(5):867–880. https://doi.org/10.1093/oxfordjournals.pcp.a076232
Nhut DT, Khai HD, Tuan NX, Bien LT, Tung HT (2022) In vitro growth and development of plants under stimulated microgravity condition. In: Nhut DT, Tung HT, Yeung EC (eds) Plant tissue culture: new techniques and application in horticultural species of tropical region. Springer, Singapore, pp 343–381
Ramage CM, Williams RR (2002) Mineral nutrition and plant morphogenesis. In Vitro Cell Dev Biol—Plant 38(2):116–124. https://doi.org/10.1079/IVP2001269
Rikiishi K, Matsuura T, Ikeda Y, Maekawa M (2015) Light inhibition of shoot regeneration is regulated by endogenous abscisic acid level in calli derived from immature barley embryos. PLoS ONE 10(12):e0145242. https://doi.org/10.1371/journal.pone.0145242
Soga K, Kurita A, Yano S, Ichikawa T, Kamada M, Takaoki M (2014) Growth and morphogenesis of azuki bean seedlings in space during SSAF2013 program. Biol Sci Space 28:6–11. https://doi.org/10.2187/bss.28.6
Soleimani M, Ghanati F, Hajebrahimi Z (2019a) The role of phenolic compounds in growth improvement of cultured tobacco cells after exposure to 2-D clinorotation. Plant Physiol 9(4):2921–2929. https://doi.org/10.30495/IJPP.2019.668858
Soleimani M, Ghanati F, Hajebrahimi Z, Hajnorouzi A, Abdolmaleki P, Zarinkamar F (2019b) Energy saving and improvement of metabolism of cultured tobacco cells upon exposure to 2-D clinorotation. J Plant Physiol 234–235:36–43. https://doi.org/10.1016/j.jplph.2019.01.002
Sutter EG, Ahmadi H, Labavitch JM (1997) Direct regeneration of Strawberry (Fragaria × ananassa Duch.) From leaf disks. Acta Hortic 447:243–246. https://doi.org/10.17660/ActaHortic.1997.447.52
Takahashi H, Kamada M, Yamazaki Y, Fujii N, Higashitani A, Aizawa S, Yoshizaki I, Kamigaichi S, Mukai C, Shimazu T, Fukui K (2000) Morphogenesis in cucumber seedlings is negatively controlled by gravity. Planta 210:515–518. https://doi.org/10.1007/s004250050039
Tang Y, Wang L, Ma C, Liu J, Liu B, Li H (2011) The use of HPLC in determination of endogenous hormones in anthers of bitter melon. J Life Sci 5:139–142
Tung HT, Thuong TT, Cuong DM, Luan VQ, Hien VT, Hieu T, Nam NB, Phuong HTN, Vinh BVT, Khai HD, Nhut DT (2021) Silver nanoparticles improved explant disinfection, in vitro growth, runner formation and limited ethylene accumulation during micropropagation of strawberry (Fragaria × ananassa). Plant Cell Tiss Org Cult 145(2):393–403. https://doi.org/10.1007/s11240-021-02015-4
Wang H, Zheng HQ, Sha W, Zeng R, Xia QC (2006) A proteomic approach to analysing responses of Arabidopsis thaliana callus cells to clinostat rotation. J Exp Bot 57(4):827–835. https://doi.org/10.1093/jxb/erj066
Wolfe K, Wu X, Liu RH (2003) Antioxidant activity of apple peels. J Agric Food Chem 51(3):609–614. https://doi.org/10.1021/jf020782a
Xie J, Zheng H (2020) Arabidopsis flowering induced by photoperiod under 3-D clinostat rotational simulated microgravity. Acta Astronaut 166:567–572. https://doi.org/10.1016/j.actaastro.2018.11.014
Xu F, Fu C, Yang Y (2020) Water affects morphogenesis of growing aquatic plant leaves. Phys Rev Lett 124(3):038003. https://doi.org/10.1103/PhysRevLett.124.038003
Zhang Z, Sun D, Zhang Y, Chen F (2020) Chloroplast morphogenesis in Chromochloris zofingiensis in the dark. Algal Res 45:101742. https://doi.org/10.1016/j.algal.2019.101742
Acknowledgements
Le The Bien was funded by the PhD Scholarship Programme of Vingroup Innovation Foundation (VINIF), code [VINIF.2022.TS013]. The authors would like to thank Prof. Edward CT Yeung (Calgary University) for critical reading of the manuscript.
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This study was supported by Vingroup Innovation Foundation (VINIF) (Grant No. VINIF.2022.TS013).
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LTB and HTT acquired data and wrote the manuscript. NTNM, THP, DMC, HDK, VQL, NBN, TTHT, BVTV participated in performing the experiments, interpretation of data and revision for intellectual content. DTN and HTT conceptualized and designed the study. All authors discussed the results and contributed to the final manuscript.
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Communicated by Maurizio Lambardi.
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Bien, L.T., Tung, H.T., Mai, N.T.N. et al. Morphogenesis of in vitro strawberry leaf cultured under clinostat 2D condition. Plant Cell Tiss Organ Cult 153, 499–510 (2023). https://doi.org/10.1007/s11240-023-02484-9
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DOI: https://doi.org/10.1007/s11240-023-02484-9