Advertisement

Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 1, pp 53–64 | Cite as

HSP70 improves the viability of cryopreserved Paeonia lactiflora pollen by regulating oxidative stress and apoptosis-like programmed cell death events

  • Ruifen Ren
  • Xueru Jiang
  • Wei Di
  • Zedi Li
  • Bingling Li
  • Jin Xu
  • Yan LiuEmail author
Original Article
  • 74 Downloads

Abstract

Heat shock proteins (HSPs) have many positive stabilizing effects on the proteins of stressed cells. This study investigated the effect of HSP70 on cryopreserved pollen. HSP70 was differentially expressed in Paeonia lactiflora and Magnolia denudate pollen before and after exposure to liquid nitrogen (LN). In this study, cryopreserved pollen samples of P. lactiflora ‘Fen Yu Nu’, ‘Zi Feng Chao Yang’ and ‘Hong Pan Tuo Jing’ were incubated with HSP70 at six concentrations, and the pollen germination, oxidative stress and apoptosis-like programmed cell death indicators were measured after LN storage. HSP70 significantly improved pollen viability after LN storage, and the concentration of HSP70 that was required varied from cultivar to cultivar, ranging from 0.5 to 10 μg/mL. Compared to the activity of the control treatment without HSP70, the activity of superoxide dismutase (SOD) increased, the contents of reactive oxygen species (ROS) and malondialdehyde (MDA) decreased, and the Ca2+ level, apoptosis rate and caspase-3-like activity decreased depending on the appropriate concentration of HSP70 added. Pollen germination was negatively correlated with MDA content, Ca2+ level, and caspase-3-like activity and positively correlated with SOD activity. In conclusion, HSP70 improved the viability of pollen after cryopreservation by improving the activity of antioxidant enzymes, reducing the contents of ROS and MDA, and affecting the Ca2+ signal to inhibit apoptosis-like programmed cell death events. This report is the first time to describe the application of HSP70 for improving cryopreserved pollen viability by regulating oxidative and apoptosis-like programmed cell death events, and provide a novel insight into the mechanisms of HSP70 action in cryopreservation.

Key Message

After the pollen was rewarmed and then incubated with appropriate concentrations of exogenous HSP70, it significantly increasing the post-freezing germination of the pollen. Which by affecting oxidative stress and apoptosis-like events, the ROS level, MDA content, intracellular Ca2+ concentration, apoptosis rate and caspase-3-like enzyme activity were reduced, and SOD activity was increased, thereby significantly increasing the post-freezing germination of the pollen.

Keywords

Cryopreservation Pollen HSP70 Oxidative stress Apoptosis-like programmed cell death events 

Notes

Acknowledgements

This research was supported by the National Natural Science Foundation of China (Nos. 31370693 and 31770741). We thank Prof. Barbara M. Reed for editing the manuscript.

Author contributions

RR designed and conducted the research, analyzed the data and drafted the manuscript. XJ and WD edited the manuscript and offered some help on experiment technical. ZL offered some help on material collection. BL and JX conceived the research and provided technical assistance. YL conceived the project, supervised the analysis and critically revised the manuscript. All authors read and approved the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Antony JJ, Keng CL, Mahmood M, Subramaniam S (2013) Effects of ascorbic acid on PVS2 cryopreservation of Dendrobium Bobby Messina’s PLBs supported with SEM analysis. Appl Biochem Biotech 171:315–329.  https://doi.org/10.1007/s12010-013-0369-x CrossRefGoogle Scholar
  2. Balogi Z, Török Z, Balogh G, Jósvay K, Shigapova N, Vierling E, Vígh L, Horváth I (2005) Heat shock lipid in cyanobacteria during heat/light-acclimation. Arch Biochem Biophys 436(2):346–354CrossRefGoogle Scholar
  3. Beere HM, Wolf BB, Cain K, Mosser DD, Mahboubi A, Kuwana T, Tailor P, Morimoto RI, Cohen GM, Green DR (2000) Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2:469–475.  https://doi.org/10.1038/35019501 CrossRefGoogle Scholar
  4. Benson EE, Bremner DH (2004) Oxidative stress in the frozen plant: a free radical point of view. In: Fuller B, Lane N, Benson EE (eds) Life in the frozen state. CRC Press, Boca Raton, pp 205–241CrossRefGoogle Scholar
  5. Chen GQ (2014) Optimization of Agapanthus embryogenic callus cryopreservation based on the Arabidopsis antioxidant mechanism. Dissertation, Shanghai Jiao Tong University (in Chinese) Google Scholar
  6. Chen GQ, Ren L, Zhang J, Reed MB, Zhang D, Shen XH (2015) Cryopreservation affects ROS-induced oxidative stress and antioxidant response in Arabidopsis seedlings. Cryobiology 70:38–47.  https://doi.org/10.1016/j.cryobiol.2014.11.004 CrossRefGoogle Scholar
  7. De Maio A (1999) Heat shock proteins: facts, thoughts, and dreams. Shock 11:1–12CrossRefGoogle Scholar
  8. Di W (2018) Study on the damage mechanism of Dendrobium nobile Lindl. ‘Hamana Lake Dream’ protocorm-like bodies during vitrification-cryopreservation. Dissertation, Beijing Forestry University (in Chinese) Google Scholar
  9. Di W, Jia MX, Liu Y (2015) Effects of exogenous antioxidants on cryopreservation of protocorm-like bodies of Dendrobium nobile. Plant Physiol J 51:1880–1886 (in Chinese) Google Scholar
  10. Di W, Jia MX, Xu J, Li BL, Liu Y (2017) Exogenous catalase and pyruvate dehydrogenase improve survival and regeneration and affect oxidative stress in cryopreserved Dedrobium nobile protocorm-like bodies. CryoLetters 28:228–238Google Scholar
  11. Engelmann F (2004) Plant cryopreservation: progress and prospects. Vitro Cell Dev Bio-Plant 40:427–433.  https://doi.org/10.1079/IVP2004541 CrossRefGoogle Scholar
  12. Goes PA, Nichi M, Silva RO, Perez EG, Dalmazzo A, Gurgel JR, Rocha C, Simoes R, Peres M, Assumpcao ME, Barnabe RC, Barnabe VH (2011) Influence of cryopreservation on the susceptibility of goat sperm against different reactive oxygen species. Reprod Fertil Dev 23:143.  https://doi.org/10.1071/RDv23n1Ab75 CrossRefGoogle Scholar
  13. Gupta MK, Uhm SJ, Lee HT (2010) Effect of vitrification and beta-mercaptoethanol on reactive oxygen species activity and in vitro development of oocytes vitrified before or after in vitro fertilization. Fertil Steril 93:2602–2607.  https://doi.org/10.1016/j.fertnstert.2010.01.043 CrossRefGoogle Scholar
  14. Guy CL, Haskell D, Li QB (1998) Association of proteins with the stress 70 molecular chaperones at low temperature: evidence for the existence of cold proteins in spinach. Cryobiology 36:301–314.  https://doi.org/10.1006/cryo.1998.2089 CrossRefGoogle Scholar
  15. Hansen G (2000) Evidence for agrobacterium-induced apoptosis in maize cells. Mol Plant-Microbe Interact 13(6):649–657.  https://doi.org/10.1094/MPMI.2000.13.6.649 CrossRefGoogle Scholar
  16. Harding K, Johnston JW, Benson EE (2009) Exploring the physiological basis of cryopreservation success and failure in clonally propagated in vitro crop plant germplasm. Agric Food Sci 18:103–116.  https://doi.org/10.2137/145960609789267524 CrossRefGoogle Scholar
  17. He GS (2014) Distribution change of Ca2+ and stress response mechanism research of Agapanthus praecox embryogenic callus during cryopreservation. Dissertation, Shanghai Jiao Tong University (in Chinese) Google Scholar
  18. Hu XL, Li YH, Yang H, Liu QJ, Li CH (2010) Heat shock protein 70 may improve the ability of antioxidant defense induced by the combination of drought and heat in Maize leaves. Acta Agronomica Sinica 36:636–644 (in Chinses) CrossRefGoogle Scholar
  19. Jeong YJ, Kim MK, Song HJ, Kang EJ, Ock SA, Kumar BM, Balasubramanian S, Rho GJ (2009) Effect of α-tocopherol supplementation during boar semen cryopreservation on sperm characteristics and expression of apoptosis related genes. Cryobiology 58:181–189.  https://doi.org/10.1016/j.cryobiol.2008.12.004 CrossRefGoogle Scholar
  20. Jia MX, Shi Y, Di W, Jiang XR, Xu J, Liu Y (2017) ROS-induced oxidative stress is closely related to pollendeterioration following cryopreservation. Vitro Cell Dev Bio-Plant 53:433–439.  https://doi.org/10.1007/s11627-017-9844-3 CrossRefGoogle Scholar
  21. Jia MX, Jiang XR, Xu J, Di W, Shi Y, Liu Y (2018) CAT and MDH improve the germination and alleviate the oxidative stress of cryopreserved Paeonia and Magnolia pollen. Acta Physiol Plant 40:37–47.  https://doi.org/10.1007/s11738-018-2612-0 CrossRefGoogle Scholar
  22. Kampfenkel K, Van MM, Inzé D (1995) Extraction and determination of ascorbate and dehydroascorbate from plant tissue. Anal Biochem 225:165–167.  https://doi.org/10.1006/abio.1995.1127 CrossRefGoogle Scholar
  23. Khomenko IP, Bakhtina LY, Zelenina OM, Kruglov SV, Manukhina EB, Bayda LA, Malyshev IY (2007) Role of heat shock proteins HSP70 and HSP32 in the protective effect of adaptation of cultured HT22 hippocampal cells to oxidative stress. Bull Exp Biol Med 144:174–177.  https://doi.org/10.1007/s10517-007-0282-9 CrossRefGoogle Scholar
  24. Kulus D, Zalewska M (2014) Cryopreservation as a tool used in long-term storage of ornamental species-a review. Sci Hortic 168:88–107.  https://doi.org/10.1016/j.scienta.2014.01.014 CrossRefGoogle Scholar
  25. Kumar RR, Goswami S, Gadpayle KA, Singh K, Sharma SK, Singh GP, Pathak H, Rai RD (2014) Ascorbic acid at pre-anthesis modulate the thermotolerance level of wheat (Triticum aestivum) pollen under heat stress. J Plant Biochem Biotechnol 23:293–306.  https://doi.org/10.1007/s13562-013-0214-x CrossRefGoogle Scholar
  26. Li BL (2010) Studies on differentially expressed protein of pollen cryopreservation and cryobank construction of Paeonia spp. Dissertation, Beijing Forestry University (in Chinese) Google Scholar
  27. Li HS, Sun Q, Zhao SJ, Zhang WH (2000) Assay of malondialdehyde in plants Experiment principle and technology of plant physiology and biochemistry. Higher Education Press, Beijing, pp 260–261 (in Chinese) Google Scholar
  28. Lin JS, Wang Y, Wang GX (2005) Salt stress-induced programmed cell death via Ca2+-mediated mitochondrial permeability transition in tobacco protoplasts. Plant Growth Regul 45:243–250.  https://doi.org/10.1007/s10725-005-5163-5 CrossRefGoogle Scholar
  29. Luza JG, Polito VS (1985) In vitro germination and storage of English walnut pollen. Sci Hortic 27:303–316.  https://doi.org/10.1016/0304-4238(85)90035-4 CrossRefGoogle Scholar
  30. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410.  https://doi.org/10.1016/S1360-1385(02)02312-9 CrossRefGoogle Scholar
  31. Mittler R (2017) ROS are good. Trends Plant Sci 22(1):11–19.  https://doi.org/10.1016/j.tplants.2016.08.002 CrossRefGoogle Scholar
  32. Mittler R, Vanderauwera S, Gollery M, Van Breuseqem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498.  https://doi.org/10.1016/j.tplants.2004.08.009 CrossRefGoogle Scholar
  33. Nguyen VT (2015) Cloning and functional analysis on heat shock protein 70 (HSP70) gene from Chimonanthus praecox (L.). Southwest University (in Chinese) Google Scholar
  34. Poobathy R, Sinniah UR, Xavier R, Subramaniam S (2013) Catalase and superoxide dismutase activities and the total protein content of protocorm-like bodies of Dendrobium Sonia-28 subjected to vitrification. Appl Biochem Biotech 170:1066–1079.  https://doi.org/10.1007/s12010-013-0241-z CrossRefGoogle Scholar
  35. Prochazkova D, Sairam RK, Srivastava GC, Singh DV (2001) Oxidative stress and antioxidant activity as the basis of senescence in maize leaves. Plant Sci 161:765–771.  https://doi.org/10.1016/S0168-9452(01)00462-9 CrossRefGoogle Scholar
  36. Qi YC, Wang HJ, Zou Y, Liu C, Liu YQ, Wang Y, Zhang W (2011) Over-expression of mitochondrial heat shock protein 70 suppresses programmed cell death in rice. FEBS Lett 585:231–239.  https://doi.org/10.1016/j.febslet.2010.11.051 CrossRefGoogle Scholar
  37. Seo JH, Naing AH, Jeon SM, Kim CK (2018) Anti-freezing-protein type III strongly influences the expression of relevant genes in cryopreserved potato shoot tips. Plant Mol Biol 97(4–5):347–355.  https://doi.org/10.1007/s11103-018-0743-8 CrossRefGoogle Scholar
  38. Shi Y (2015) Study on oxidative stress of the pollen of ornamental plants during cryopreservation. Dissertation, Beijing Forestry University (in Chinese) Google Scholar
  39. Skyba M, Petijová L, Košuth J, Koleva DP, Ganeva TG, Kapchina-Toteva VM, Cellárová E (2012) Oxidative stress and antioxidant response in Hypericum perforatum L. plants subjected to low temperature treatment. J Plant Physiol 169:955–964.  https://doi.org/10.1016/j.jplph.2012.02.017 CrossRefGoogle Scholar
  40. Stankiewicz AR, Lachapelle G, Foo CPZ, Radicioni SM, Mosser DD (2005) Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 280:38729–38739.  https://doi.org/10.1074/jbc.M509497200 CrossRefGoogle Scholar
  41. Thomas SG, Franklin-Tong V (2004) Self-incompatibility triggers programmed cell death in Papaver pollen. Nature 429:305–309.  https://doi.org/10.1038/nature02540 CrossRefGoogle Scholar
  42. Tian AG, Wang F, Zhang W, Liu CM, Zhao XM (2012) Antioxidant mechanism and lipid peroxidation patterns in leaves and petals of Marigold in response to drought stress. Hortic Environ Biotechnol 53:183–192.  https://doi.org/10.1007/s13580-012-0069-4 CrossRefGoogle Scholar
  43. Uchendu EE, Leonard SW, Traber MG, Reed BM (2010) Vitamins C and E improve regrowth and reduce lipid peroxidation of blackberry shoot tips following cryopreservation. Plant Cell Rep 29:25–35.  https://doi.org/10.1007/s00299-009-0795-y CrossRefGoogle Scholar
  44. Varghese T, Divyashree BC, Roy SC, Roy KS (2016) Loss of heat shock protein 70 from apical region of buffalo (Bubalus bubalis) sperm head after freezing and thawing. Theriogenology 85(5):828–834CrossRefGoogle Scholar
  45. Vendrell-Flotats M, Arcarons N, Barau E, López-Béjar M, Mogas T (2017) Effect of heat stress during in vitro maturation on developmental competence of vitrified bovine oocytes. Reprod Domest Anim 52:48–51.  https://doi.org/10.1111/rda.13055 CrossRefGoogle Scholar
  46. Volk GM (2010) Application of functional genomics and proteomics to plant cryopreservation. Curr Genomics 11:24–29.  https://doi.org/10.2174/138920210790217945 CrossRefGoogle Scholar
  47. Wang CR, Wang XR, Tian Y, Xue YG, Xu XH, Sui YX, Yu HX (2008) Oxidative stress and potential biomarkers in tomato seedlings subjected to soil lead contamination. Ecotoxicol Environ Saf 71:685–691.  https://doi.org/10.1016/j.ecoenv.2008.01.002 CrossRefGoogle Scholar
  48. Wen B, Wang R, Cheng H, Song S (2010) Cytological and physiological changes in orthodox maize embryos during cryopreservation. Protoplasma 239:57–67.  https://doi.org/10.1007/s00709-009-0083-2 CrossRefGoogle Scholar
  49. Wen B, Cai C, Wang R, Song S, Song J (2012) Cytological and physiological changes in recalcitrant chinese fanpalm (Livistona chinensis) embryos during cryopreservation. Protoplasma 249:323–335.  https://doi.org/10.1007/s00709-011-0283-4 CrossRefGoogle Scholar
  50. Whitaker C, Beckett RP, Minibayeva FV, Kranner I (2010) Production of reactive oxygen species in excised, desiccated and cryopreserved explants of Trichilia dregeana Sond. S Afr J Bot 76:112–118.  https://doi.org/10.1016/j.sajb.2009.09.008 CrossRefGoogle Scholar
  51. Wu YL (2011) Cryopreservation of Dendrobium wardianum Warner. protocorms by vitrification. Dissertation, Shanghai Jiao Tong University (in Chinese) Google Scholar
  52. Xu J (2014) A study on the mechanism of Magnolia denudata pollen cryopreservation. Dissertation, Beijing Forestry University (in Chinese) Google Scholar
  53. Xu J, Liu Q, Jia MX, Liu Y, Li BL, Shi Y (2014) Generation of reactive oxygen species during cryopreservation may improve Lilium × siberia pollen viability. Vitro Cell Dev Bio-Plant 50:369–375.  https://doi.org/10.1007/s11627-014-9615-3 CrossRefGoogle Scholar
  54. Zhang YL (2007) Pollen cryopreservation of Prunus mume Sieb. et. Zuce. and the conslruction of pollen bank, Dissertation, Beijing Forestry University (in Chinese) Google Scholar
  55. Zhang J (2015) Influence mechanism of ROS induced oxidative stress and apoptosis on cell viability of Agapanthus praecox callus during cryopreservation. Dissertation, Shanghai Jiao Tong University (in Chinese) Google Scholar
  56. Zhang HY, Wang WX, Yin H, Zhao XM, Du YG (2012) Oligochitosan induces programmed cell death in tobacco suspension cells. Carbohydr Polym 87:2270–2278.  https://doi.org/10.1016/j.carbpol.2011.10.059 CrossRefGoogle Scholar
  57. Zhang D, Ren L, Chen GQ, Zhang J, Reed BM, Shen XH (2015) ROS-induced oxidative stress and apoptosis-like event directly affect the cell viability of cryopreserved embryogenic callus in Agapanthus praecox. Plant Cell Rep 34:1499–1513.  https://doi.org/10.1007/s00299-015-1802-0 CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Beijing Laboratory of Urban and Rural Ecological Environment, Beijing Municipal Education Commission, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, National Engineering Research Center for Floriculture, College of Landscape ArchitectureBeijing Forestry UniversityBeijingChina
  2. 2.College of Architecture and Urban PlanningGuangzhou UniversityGuangzhouChina

Personalised recommendations