Skip to main content
Log in

Programmed cell death induced by cadmium stress and/or boron deprivation in tobacco (Nicotiana tabacum L.) cultivar Bright Yellow 2 cells

  • Cell Biology
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Cadmium (Cd) is one of the most toxic and widespread heavy metal pollutants in soil. As an essential mineral nutrient, boron (B) plays critical roles in physiological processes of plants. In the present study, programmed cell death (PCD) induced by Cd stress and/or B deprivation was assessed and the underlying mechanisms were clarified in suspension-cultured Nicotiana tabacum L. cultivar Bright Yellow 2 (TBY-2) cells. The PCD in TBY-2 cells was analyzed by Hoechst 33258 staining and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and then, expression analysis of PCD-related genes was performed using quantitative real-time polymerase chain reaction (qPCR) assays. The production of reactive oxygen species (ROS) was determined using fluorescence microscopy of 2′,7′-dichlorofluorescein diacetate–labeled cells. The levels of lipid peroxides were quantified by the thiobarbituric acid–reactive substances (TBARS) method. Cadmium stress and/or B deprivation treatments induced PCD that was characterized by a significant increase in the percentage of cells stained with Hoechst 33258 or TUNEL-positive cells, and upregulation or downregulation of the expression of PCD-related genes. Treatments with Cd stress and/or B deprivation increased ROS production and the level of lipid peroxides compared to those of the control group. These data showed that in TBY-2 cells Cd stress and/or B deprivation activated ROS signaling pathways, leading to gene expression that was connected with the PCD process.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Anatawirron P, Subedi KD, Rerkasem B (1997) Screening wheat for boron efficiency. In: Bell RW, Rerkasem B (eds) Boron in soils and plants. Kluwer Academic Publishers, The Netherlands, pp 101–104

    Chapter  Google Scholar 

  • Bi Y, Chen W, Zhang W, Zhou Q, Yun L, Xing D (2009) Production of reactive oxygen species, impairment of photosynthetic function and dynamic changes in mitochondria are early events in cadmium induced cell death in Arabidopsis thaliana. Biol Cell 101:629–643

    Article  PubMed  CAS  Google Scholar 

  • Burbridge E, Diamond M, Dix PJ, McCabe PF (2007) Use of cell morphology to evaluate the effect of a peroxidase gene on the cell death induction thresholds in tobacco. Plant Sci 172:853–860

    Article  CAS  Google Scholar 

  • Chuaqui RF, Bonner RF, Best CJM, Gillespie JW, Flaig MJ, Hewitt SM, Phillips JL, Krizman DB, Tangrea MA, Ahram M, Linehan WM, Knezevic V, Emmert- Buck MR (2002) Post-analysis follow-up and validation of microarray experiments. Nature Gen 32(Suppl):509–514

    Article  CAS  Google Scholar 

  • Das P, Samantaray S, Rout G (1997) Studies on cadmium toxicity in plants: a review. Environ Pollut 98:29–36

    Article  PubMed  CAS  Google Scholar 

  • de Michele R, Vurro E, Rigo C, Costa A, Elviri L, Di Valentin M, Careri M, Zottini M, Sanità di Toppi L, Lo Schiavo F (2009) Nitric oxide is involved in cadmium-induced programmed cell death in Arabidopsis suspension cultures. Plant Physiol 150:217–228

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • de Oliveira LA, Breton MC, Bastolla FM, Camargo Sda S, Margis R, Frazzon J, Pasquali G (2012) Reference genes for the normalization of gene expression in eucalyptus species. Plant Cell Physiol 53:405–422

    Article  PubMed  CAS  Google Scholar 

  • de Pinto MC, Francis D, De Gara L (1999) The redox state of the ascorbate–dehydroascorbate pair as a specific sensor of cell division in tobacco BY-2 cells. Protoplasma 209:90–97

    Article  PubMed  Google Scholar 

  • de Pinto MC, Paradiso A, Leonetti P, De Gara L (2006) Hydrogen peroxide, nitric oxide and cytosolic ascorbate peroxidase at the crossroad between defense and cell death. Plant J 48:784–795

    Article  PubMed  CAS  Google Scholar 

  • di Toppi LS, Gabbrielli R (1999) Response to cadmium in higher plants. Environ Exp Bot 41:105–130

    Article  Google Scholar 

  • Ferradás Y, López M, Rey M, González MV (2014) Programmed cell death in kiwifruit stigmatic arms and its relationship to the effective pollination period and the progamic phase. Ann Bot 114:35–45

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Foyer CH, Noctor G (2005) Oxidant and antioxidant signaling in plants: a re-evaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ 29:1056–1071

    Article  Google Scholar 

  • Garnier L, Simon-Plas F, Thuleau P, Agnel JP, Blein JP, Ranjeva R, Montillet JL (2006) Cadmium affects tobacco cells by a series of three waves of reactive oxygen species that contribute to cytotoxicity. Plant Cell Environ 29:1956–1969

    Article  PubMed  CAS  Google Scholar 

  • Giordani C, Cecchi S, Zanchi C (2005) Phytoremediation of soil polluted by nickel using agricultural crops. Environ Manag 36:675–681

    Article  Google Scholar 

  • Gunawardena AH (2008) Programmed cell death and tissue remodeling in plants. J Exp Bot 59:445–451

    Article  PubMed  CAS  Google Scholar 

  • Heath MC (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–324

    Article  PubMed  CAS  Google Scholar 

  • Iakimova ET, Woltering EJ, Kapchina-Toteva VM, Harren FJM, Cristescu SM (2008) Cadmium toxicity in cultured tomato cells—role of ethylene, proteases and oxidative stress in cell death signaling. Cell Biol Int 32:1521–1529

    Article  PubMed  CAS  Google Scholar 

  • Ikenaka Y, Nakayama SMM, Muzandu K, Choongo K, Teraoka H, Mizuno N, Ishizuka M (2010) Heavy metal contamination of soil and sediment in Zambia. Afr J Environ Sci Technol 4:729–739

    CAS  Google Scholar 

  • Kenneth JL, Thomas DS (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. 25:402–408

  • Kobayashi M, Mutoh T, Matoh T (2004) Boron nutrition of cultured tobacco BY-2 cells. IV. Genes induced under low boron supply. J Exp Bot 55:1441–1443

    Article  PubMed  CAS  Google Scholar 

  • Koshiba T, Kobayashi M, Ishihara A, Matoh T (2010) Boron nutrition of cultured tobacco BY-2 cells. VI. Calcium is involved in early responses to boron deprivation. Plant Cell Physiol 51:323–327

    Article  PubMed  CAS  Google Scholar 

  • Koshiba T, Kobayashi M, Matoh T (2009) Boron nutrition of tobacco BY-2 cells. V. Oxidative damage is the major cause of cell death induced by boron deprivation. Plant Cell Physiol 50:26–36

    Article  PubMed  CAS  Google Scholar 

  • Kutik J, Kuthanova A, Smertenko A, Fischer L, Opatrny Z (2014) Cadmium-induced cell death in BY-2 cell culture starts with vacuolization of cytoplasm and terminates with necrosis. Physiol Plant 151:423–433

    Article  PubMed  CAS  Google Scholar 

  • Lachaud C, Da Silva D, Amelot N, Béziat C, Brière C, Cotelle V, Graziana A, Grat S, Mazars C, Thuleau P (2011) Dihydrosphingosine-induced programmed cell death in tobacco BY-2 cells is independent of H2O2 production. Mol Plant 4:310–318

    Article  PubMed  CAS  Google Scholar 

  • Liu D, Jiang W, Zhang L, Li L (2000) Effects of boron ions on root growth and cell division of broadbean (Vicia faba L.). Israel J Plant Sci 48:47–51

    Article  CAS  Google Scholar 

  • Lockshin RA, Zakeri Z (2004) Apoptosis, autophagy, and more. Int J Biochem Cell Biol 36:2405–2419

    Article  PubMed  CAS  Google Scholar 

  • Ma W, Xu W, Xu H, Chen Y, He Z, Ma M (2010) Nitric oxide modulates cadmium influx during cadmium-induced programmed cell death in tobacco BY-2 cells. Planta 232:325–335

    Article  PubMed  CAS  Google Scholar 

  • Moran JF, Becana M, Iturbe-Ormaetxe I, Frechilla S, Klucas RV, Aparicio-Tejo P (1994) Drought induces oxidative stress in pea plants. Planta 194:346–352

    Article  CAS  Google Scholar 

  • Mou Z, Fan W, Dong X (2003) Inducers of plant systemic acquired resistance regulate NPR1 function through redox changes. Cell 113:935–944

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Myouga F, Hosoda C, Umezawa T, Iizumi H, Kuromori T, Motohashi R, Shinozaki K (2008) A heterocomplex of superoxide dismutases defends chloroplast nucleoids against oxidative iron stress and is essential for chloroplast development in Arabidopsis. Plant Cell 20:3148–3162

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Murata Y, Obi I, Yoshihashi M, Noguchi M, Kakutani T (1994) Reduced permeability to K+ and Na+ ions on K+ channels in the plasma membrane of tobacco cells in suspension after adaptation to 50 mM NaCl. Plant Cell Physiol 35:87–92

    Article  CAS  Google Scholar 

  • Olmos E, Martinez-Solano JR, Piqueras A, Hellin E (2003) Early steps in the oxidative burst induced by cadmium in cultured tobacco cells (BY-2 line). J Exp Bot 54:291–301

  • Pontier D, Gan S, Amasino RM, Roby D, Lam E (1999) Markers for hypersensitive response and senescence show distinct patterns of expression. Plant Mol Bio 39:1243–1255

    Article  CAS  Google Scholar 

  • Reape TJ, Molony EM, McCabe PF (2008) Programmed cell death in plants: distinguishing between different modes. J Exp Bot 59:435–444

    Article  PubMed  CAS  Google Scholar 

  • Rodríguez-Serrano M, Bárány I, Prem D, Coronado MJ, Risueño MC, Testillano PS (2012) NO, ROS, and cell death associated with caspase-like activity increase in stress-induced microspore embryogenesis of barley. J Exp Bot 63:2007–2024

    Article  PubMed  CAS  Google Scholar 

  • Rogers HJ (2006) Programmed cell death in floral organs: how and why do flowers die? Ann Bot 97:309–315

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Rustérucci C, Montillet JL, Agnel JP, Battesti C, Alonso B, Knoll A, Bessoule JJ, Etienne P, Suty L, Blein JP, Triantaphylidès C (1999) Involvement of lipoxygenase-dependent production of fatty acid hydroperoxides in the development of the hypersensitive cell death induced by cryptogein on tobacco leaves. J Bio Chem 274:36446–36455

    Article  Google Scholar 

  • Schützendübel A, Polle A (2002) Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization. J Exp Bot 53:1351–1365

    PubMed  Google Scholar 

  • Schwarzerová K, Zelenková S, Nick P, Opatrný Z (2002) Aluminum-induced rapid changes in the microtubular cytoskeleton of tobacco cell lines. Plant Cell Physiol 43:207–216

    Article  PubMed  Google Scholar 

  • Sewalt VJH, Ni W, Jung HG, Dixon RA (1997) Lignin impact on fiber degradation: increased enzymatic digestibility of genetically engineered tobacco (Nicotiana tabacum) stems reduced in lignin content. J Agr Food Chem 45:1977–1983

    Article  CAS  Google Scholar 

  • Shi HT, Chan ZL (2014) Improvement of plant abiotic stress tolerance through modulation of the polyamine pathway. J Integr Plant Biol 56:114–121

    Article  PubMed  CAS  Google Scholar 

  • Shorrocks VM (1997) The occurrence and correction of boron deficiency. Plant Soil 193:121–148

    Article  CAS  Google Scholar 

  • Siedlecka A, Krupa Z (1996) Interaction between cadmium and iron and its effects on photosynthetic capacity of primary leaves of Phaseolus vulgaris. Plant Physiol Bioch 34:833–841

    CAS  Google Scholar 

  • Sytar O, Kumar A, Latowski D, Kuczynska P, Strzałka K, Prasad MNV (2013) Heavy metal-induced oxidative damage, defense reactions, and detoxification mechanisms in plants. Acta Physiol Plant 35:985–999

    Article  CAS  Google Scholar 

  • Tewari RK, Kumar P, Sharma PN (2009) Morphology and oxidative physiology of boron-deficient mulberry plants. Tree Physiol 30:68–77

    Article  PubMed  CAS  Google Scholar 

  • Vacca RA, de Pinto MC, Valenti D, Passarella S, Marra E, De Gara L (2004) Production of reactive oxygen species, alteration of cytosolic ascorbate peroxidase, and impairment of mitochondrial metabolism are early events in heat shock-induced programmed cell death in tobacco bright-yellow 2 cells. Plant Physiol 134:1100–1112

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • van Doorn WG, Woltering EJ (2005) Many ways to exit? Cell death categories in plants. Trends Plant Sci 10:117–122

    Article  PubMed  CAS  Google Scholar 

  • Wang Q (1998) In integrated amendment and ecological restoration of polluted soil by heavy metals. Proceedings for strategy of soil environmental protection in the new century of China. Oct, Beijing, pp 26–28

    Google Scholar 

  • Wang Q, Li J (1999) Fertilizer proper use and sustainable development of soil environment in China. Adv Environ Sci 7:116–124

    Google Scholar 

  • Wen F, Xing D, Zhang LR (2008) Hydrogen peroxide is involved in high blue light-induced chloroplast avoidance movements in Arabidopsis. J Exp Bot 59:2891–2901

    Article  PubMed  CAS  Google Scholar 

  • Yabuta Y, Maruta T, Yoshimura K, Ishikawa T, Shigeoka S (2004) Two distinct redox signaling pathways for cytosolic APX induction under photooxidative stress. Plant Cell Physiol 45:1586–1594

    Article  PubMed  CAS  Google Scholar 

  • Ye Y, Li Z, Xing D (2012) Sorting out the role of nitric oxide in cadmium-induced Arabidopsis thaliana programmed cell death. Plant Signal Behav 7:1493–1494

    Article  PubMed  PubMed Central  Google Scholar 

  • Ye Y, Li Z, Xing D (2013) Nitric oxide promotes MPK6-mediated caspase-3-like activation in cadmium-induced Arabidopsis thaliana programmed cell death. Plant Cell Environ 36:1–15

    Article  PubMed  CAS  Google Scholar 

  • Yu Q, Hlavacka A, Matoh T, Volkmann D, Menzel D, Goldbach HE, Baluska F (2002) Short-term boron deprivation inhibits endocytosis of cell wall pectins in meristematic cells of maize and wheat root apices. Plant Physiol 130:415–421

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhang L, Wong MH (2007) Environmental mercury contamination in China: sources and impacts. Environ Int 33:108–121

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The Liaoning Key Laboratory for Ecologically Comprehensive Utilization of Boron Resources and Materials, Northeastern University, Liaoning, Shenyang, China, supported this work.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lai Fu Zhong.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Editor: Tom Clemente

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jiang, L.P., Zi, L., Liu, X.F. et al. Programmed cell death induced by cadmium stress and/or boron deprivation in tobacco (Nicotiana tabacum L.) cultivar Bright Yellow 2 cells. In Vitro Cell.Dev.Biol.-Plant 55, 15–25 (2019). https://doi.org/10.1007/s11627-019-09960-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-019-09960-y

Keywords

Navigation