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Changes in endogenous hormones and H2O2 burst during shoot organogenesis in TDZ-treated Saussurea involucrate explants

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Abstract

Thidiazuron [N-phenyl-N-(1, 2, 3-thidiazol-5-yl) urea, TDZ] treatment significantly improved shoot morphogenesis of Saussurea involucrata Kar. Et Kir (S. involucrata) leaf explants. The biochemical mechanisms underlying TDZ-induced shoot organogenesis were investigated by measuring endogenous plant growth hormones, H2O2, as well as the activities of superoxide dismutase (SOD) and catalase (CAT). The levels of endogenous gibberellic A3 (GA3) and zeatin (ZT) significantly increased in leaf explants subject to a 28-day treatment than the controls. However, extending exposure time to TDZ inhibited GA3 accumulation. At the same time, the SOD activity increased significantly until the 28th of TDZ treatment time and the CAT activity reduced simultaneously, which was closely linked with the significant increase in H2O2 concentrations in the explants. And there was a sharply promotion after the 35 day of culture time if the plant tissue was always in medium contained, which was in company with the cell activity decreased. We propose that a combination of increased GA3, ZT, and H2O2 concentration is the basis for the enhanced shoot morphogenesis in response to TDZ treatment. These results provide a starting point for an improved understanding of the biochemical mechanisms underlying TDZ-induced shoot organogenesis of S. involucrata.

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References

  • Abbasi BH, Khan M, Guo B, Bokhari SA, Khan MA (2011) Efficient regeneration and antioxidative enzyme activities in Brassica rapa var. Turnip. Plant Cell Tissue Organ Cult 105:337–344

    Article  CAS  Google Scholar 

  • Ali M, Abbasi BH (2014) Thidiazuron-Induced changes in biomass parameters, total phenolic content, and antioxidant activity in callus cultures of Artemisia absinthium L. Appl Biochem Biotechnol 172:2363–2376

    Article  CAS  PubMed  Google Scholar 

  • Alscher RG, Donahue JL, Cramer CL (1997) Reactive oxygen species and antioxidants: relationships in green cells. Physiol Plant 100:224–233

    Article  CAS  Google Scholar 

  • Aremu AO, Plačková L, Bairu MW, Novák O, Plíhalová L, Doleźal K, Finnie JF, Van Staden J (2014a) How does exogenously applied cytokinin type affect growth and endogenous cytokinins in micropropagated Merwilla plumbea? Plant Cell Tissue Organ Cult 118:245–256

  • Aremu AO, Plačková L, Bairu MW, Novák O, Szüčová L, Doležal K, Finnie JF, Van Staden J (2014b) Endogenous cytokinin profiles of tissue-cultured and acclimatized ‘Williams’ bananas subjected to different aromatic cytokinin treatments. Plant Sci 214:88–98

  • Arshad M, Silvestre J, Merlina G, Dumat C, Pinelli E, Kallerhoff J (2012) Thidiazuron-induced shoot organogenesis from mature leaf explants of scented Pelargonium capitatum cultivars. Plant Cell Tissue Organ Cult 108(2):315–322

    Article  CAS  Google Scholar 

  • Bakshi S, Roy NK, Sahoo L (2012) Seedling preconditioning in thidiazuron enhances axillary shoot proliferation and recovery of transgenic cowpea plants. Plant Cell Tissue Organ Cult 110(1):77–91

    Article  CAS  Google Scholar 

  • Beaucham C, Fridovich I (1971) Superoxide dismutase—improved assays and an assay applicable to acrylamide gels. Anal Biochem 44(1):276–287

    Article  Google Scholar 

  • Bright J, Desikan R, Tancock JT, Weir IS, Neill SJ (2006) ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis. Plant J 45:113–122

    Article  CAS  PubMed  Google Scholar 

  • Casanova E, Valdes AE, Fernandez B, Moysset L, Trillas MI (2004) Levels and immunolocalization of endogenous cytokinins in thidiazuron-induced shoot organogenesis in carnation. J Plant Physiol 161(1):95–104

    Article  CAS  PubMed  Google Scholar 

  • Chhabra G, Chaudhary D, Varma M, Sainger M, Jaiwal PK (2008) TDZ-induced direct shoot organogenesis and somatic embryogenesis on cotyledonary node explants of lentil (Lens culinaris Medik.). Physiol Mol Biol Plants 14(4):347–353

    Article  CAS  PubMed  Google Scholar 

  • Corredoira E, Ballester A, Vieitez AM (2008) Thidiazuron-induced high-frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees. Plant Cell Tissue Organ Cult 95:197–208

    Article  CAS  Google Scholar 

  • Deroles SC, Seelye JF, John J, Mullan AC (2011) In vitro propagation of Sandersonia aurantiaca Hook using thidiazuron. Plant Cell Tissue Organ Cult 102:115–119

    Article  Google Scholar 

  • Fu LG (1992) China plant red data book-rare and endangered plants, vol 1. Chinese Science Press, Beijing, pp 234–235

    Google Scholar 

  • Guo ZJ, Liu LY, Zhang WY, Zang UG (2006) Resource condirions, management and conservation strategies of snow lotus (Saussurea involucrate) in Tianshan mountains. J Arid Land Resour Environ 2:97–103

    Google Scholar 

  • Guo B, Gao M, Liu CZ (2007) In vitro propagation of an endangered medicinal plant Saussurea involucrata Kar. et Kir. Plant Cell Rep 26:261–265

    Article  CAS  PubMed  Google Scholar 

  • Guo B, Stiles Amanda R, Liu C (2012) Thidiazuron enhances shoot organogenesis from leaf explants of Saussurea involucrata Kar. et Kir. In Vitro Cell Dev Biol Plant 48(6):609–612

    Article  CAS  Google Scholar 

  • Guo B, Stiles Amanda R, Liu C (2013) Changes in endogenous hormones and oxidative burst as the biochemical basis for enhanced shoot organogenesis in cold-treated Saussurea involucrata explants. Acta Physiol Plant 35:283–287

    CAS  Google Scholar 

  • Gupta SD (2011) Role of free radicals and antioxidants in in vitro morphogenesis. In: Reactive oxygen species and antioxidants in higher plants. Science Publishers, Enfield, pp 229–247

  • Huang WL, Lee CH, Chen YR (2012) Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress. Plant Cell Tissue Organ Cult 108:257–263

    Article  CAS  Google Scholar 

  • Jia JM, Wu CF, Liu W, Yu H, Hao Y, Zheng JH, Ji YR (2005) Antiinflammatory and analgesic activities of the tissue culture of Saussurea involucrate. Biol Pharm Bull 28:1612–1614

    Article  CAS  PubMed  Google Scholar 

  • Li GH, Zhao RC (1989) Studies on pharmacological actions of Saussure involucrata Kar. et Kir. Acta Pharm Sin 15:368–369

    Google Scholar 

  • Liu L, Xiao X, Zhang L (1985) Effect of the flavonoids from Saussurea involucrata on DNA synthesis of cancer cells. Lanzhou Univ Nat Sci 21:80–83

    CAS  Google Scholar 

  • Liu CZ, Murch SJ, EL-Demerdash M (2003) Regeneration of the Egyptian medicinal plant Artemisia judaica L. Plant Cell Rep 21:525–530

    CAS  PubMed  Google Scholar 

  • Liu CZ, Murch SJ, Jain JC, Saxena PK (2004a) Goldenseal (Hydrastis canadensis L.): in vitro regeneration for germplasm conservation and elimination of heavy metal contamination. In Vitro Cell Dev Biol Plant 40:75–79

  • Lu SY, Zhuo CL, Wang XH, Guo ZF (2014) Nitrate reductase (NR)-dependent NO production mediates ABA- and H2O2-induced antioxidant enzymes. Plant Physiol Biochem 74:9–15

    Article  CAS  PubMed  Google Scholar 

  • Murch SJ, Saxena PK (2001) Molecular fate of thidiazuron and its effects on auxin transport in hypocotyls tissues of Pelargonium × hortorum Bailey. Plant Growth Regul 35(3):269–275

    Article  CAS  Google Scholar 

  • Murch SJ, Saxena PK (2007) Thidiazuron-induced regeneration of Echinacea purpurea L: Micropropagation in solid and liquid culture systems. Plant Cell Rep 26:13–19

    PubMed  Google Scholar 

  • Murch SJ, Victor JMR, Krishnaraj S, Saxena PK (1999) The role of proline in thidiazuron-induced somatic embryogenesis of peanut. In Vitro Cell Dev Biol 35:102–105

    Article  CAS  Google Scholar 

  • Murthy BNS, Murch SJ, Saxena PK (1995) TDZ-induced somatic embryogenesis in intact seedlings of peanut (Arachis hypogaea): endogenous growth regulator levels and significance of cotyledons. Physiol Plant 94:268–276

    Article  CAS  Google Scholar 

  • Murthy BNS, Murch SJ, Saxena PK (1998) Thidiazuron: a potent regulator of in vitro plant morphogenesis. In Vitro Cell Dev Biol Plant 34:267–275

    Article  CAS  Google Scholar 

  • Scandalios JG (1993) Oxygen stress and superoxide dismutases. Plant Physiol 101(1):7–12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Souza BM, Kraus JE, Endres L, Mercier H (2003) Relationships between endogenous hormonal levels and axillary bud development of Ananas comosus nodal segments. Plant Physiol Biochem 41:733–739

    Article  CAS  Google Scholar 

  • Steponkus PL, Lanphear FO (1967) Refinement of triphenyl tetrazolium chloride method of determining cold injury. Plant Physiol 42:1423–1426

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Suzuki RM, Kerbauy GB, Zaffari GR (2004) Endogenous hormonal levels and growth of dark-incubated shoots of Catasetum fimbriatum. Plant Physiol 161:929–935

    Article  CAS  Google Scholar 

  • Tang W, Newton RJ (2005) Peroxidase and catalase activities are involved in direct adventitious shoot formation induced by thidiazuron in Eastern white pine (Pinus strobus L.) zygotic embryos. Plant Physiol Biochem 43(8):760–769

    Article  CAS  PubMed  Google Scholar 

  • Tatjana C, Vaclav M, Martin R, Jelena S, Aleksandar C, Branka V, Dragan V, Alena T, Petre ID, Borut B, Slavica N (2015) In vitro shoot organogenesis and comparative analysis of endogenous phytohormones in kohlrabi (Brassica oleracea var. gongylodes): effects of genotype, explant type and applied cytokinins. Plant Cell Tissue Organ Cult 121:741–760

    Article  Google Scholar 

  • Tian M, Gu Q, Zhu MY (2003) The involvement of hydrogen peroxide and antioxidant enzymes in the process of shoot organogenesis of strawberry callus. Plant Sci 165(4):701–707

    Article  CAS  Google Scholar 

  • Wang A, Luo G (1990) Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants. Plant Physiol Commun 6:55

    CAS  Google Scholar 

  • Wei SJ, Wu YF, Luo YY, Ke X, Yang L (2014) Advances in studies on the resource of an endangered Chinese medicinal herb of Saussurea involucrate. J MUC (Nat Sci Ed) 23(2):10–15

    Google Scholar 

  • Xu J, Yin H, Li Y, Liu X (2010) Nitric oxide is associated with long-term zinc tolerance in Solanum nigrum. Plant Physiol 154:1319–1334

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang CG, Li W, Mao YF, Zhao DL, Dong W, Guo GQ (2005) Endogenous hormonal levels in scutellaria baicalensis calli induced by thidiazuron. Russ J Plant Physiol 52(3):345–351

    Article  CAS  Google Scholar 

  • Zhang YF, Zhou JH, Wu T, Cao JS (2008) Shoot regeneration and the relationship between organogenic capacity and endogenous hormonal contents in pumpkin. Plant Cell Tissue Organ Cult 93:323–331

    Article  Google Scholar 

  • Zhang W, Wang XM, Fan R, Yin GX, Wang K, Du LP, Xiao LL, Ye XG (2015) Effects of inter-culture, arabinogalactan proteins, and hydrogen peroxide on the plant regeneration of wheat immature embryos. J Integr Agric 14(1):11–19

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by Special Fund for Agro-scientific Research in The Public Interest (Grant No. 201203062), Natural Science Foundation of China (Grant Nos. 31402133, 41303063, 31572665, 30970308), the China Postdoctoral Science Foundation (Grant No. 2016M592830), Special Fund for Scientific Research from Shaanxi Provincial Department of Education (Grant No. 14JK1725), and Project of Natural Science Foundation research project of Shaanxi Province (Grant No. 2016JM3002).

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Correspondence to Yahui Wei.

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Bin Guo and Wei He have contributed equally to this work.

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Guo, B., He, W., Zhao, Y. et al. Changes in endogenous hormones and H2O2 burst during shoot organogenesis in TDZ-treated Saussurea involucrate explants. Plant Cell Tiss Organ Cult 128, 1–8 (2017). https://doi.org/10.1007/s11240-016-1069-3

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