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Survival and ultrastructural features of peach palm (Bactris gasipaes, Kunth) somatic embryos submitted to cryopreservation through vitrification

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Abstract

Bactris gasipaes (Arecaceae), also known as peach palm, was domesticated by Amazonian Indians and is cultivated for its fruit and heart-of-palm, a vegetable grown in the tree’s inner core. Currently, the conservation of this species relies on in situ conditions and field gene banks. Complementary conservation strategies, such as those based on in vitro techniques, are indicated in such cases. To establish an appropriate cryopreservation protocol, this study aimed to evaluate the ultrastructural features of B. gasipaes embryogenic cultures submitted to vitrification and subsequent cryogenic temperatures. Accordingly, somatic embryo clusters were submitted to Plant Vitrification Solution 3 (PVS3). In general, cells submitted to PVS3 had viable cell characteristics associated with apparently many mitochondria, prominent nucleus, and preserved cell walls. Cells not incubated in PVS3 did not survive after the cryogenic process in liquid nitrogen. The best incubation time for the vitrification technique was 240 min, resulting in a survival rate of 37 %. In these cases, several features were indicative of quite active cell metabolism, including intact nuclei and preserved cell walls, an apparently many of mitochondria and lipid bodies, and the presence of many starch granules and condensed chromatin. Moreover, ultrastructure analysis revealed that overall cellular structures had been preserved after cryogenic treatment, thus validating the use of vitrification in conjunction with cryopreservation of peach palm elite genotypes, as well as wild genotypes, which carry a rich pool of genes that must be conserved.

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Abbreviations

CBB:

Coomassie Brilliant Blue

DMSO:

Dimethyl sulfoxide

FM:

Fresh mass

LN:

Liquid nitrogen

MS:

Murashige and Skoog

Picloram:

4-Amino-3,5,6-trichloropicolinic acid

PAS:

Periodic Acid-Schiff

PVS:

Plant vitrification solution

SEC:

Somatic embryo clusters

SNK:

Student–Newman–Keuls

TEM:

Transmission electron microscopy

TIS:

Temporary immersion system

References

  • Al-Bahrany AM, Al-Khayri JM (2012) Optimizing in vitro cryopreservation of date palm (Phoenix dactylifera L.). Biotechnol 11:59–66

    Article  CAS  Google Scholar 

  • Benson EE (2004) Cryo-conserving algal and plant diversity: historical perspectives and future challenge. In: Fuller B, Lane N, Benson EE (eds) Life in the frozen state. CRC, London, pp 299–328

    Chapter  Google Scholar 

  • Benson EE (2008) Cryopreservation theory. In: Reed BM (ed) In plant cryopreservation: a practical guide. Springer, Heidelberg

    Google Scholar 

  • Benson EE, Bremner D (2004) Oxidative stress in the frozen plant: a free radical point of view. In: Fuller BJ, Lane N, Benson EE (eds) Life in the frozen state. CRC, Boca Raton, pp 205–241

    Chapter  Google Scholar 

  • Bovi MLA, Martins CC, Spiering SH (2004) Desidratação de sementes de quatro lotes de pupunheira: efeitos sobre a germinação e o vigor. Hortic Bras 22:109–112

    Article  Google Scholar 

  • Clement CR, Mora-Urpí JE (1987) Pejibaye palm (Bactris gasipaes, Arecaceae): multi-use potential for the lowland humid tropics. Econ Bot 41:302–311

    Article  Google Scholar 

  • Clement CR, Santos RP, Desmouliere SJM, Ferreira EJL, Neto JTF (2009) Ecological adaptation of wild peach palm, its in situ conservation and deforestation-mediated extinction in southern Brazilian Amazonia. PLoS One 4(2):e4564

    Article  PubMed  Google Scholar 

  • Engelmann F (2000) Importance of cryopreservation for the conservation of plant genetic resources. In: Engelmann F, Takagi H (eds) Cryopreservation of tropical plant germplasm: current research progress and application. IPGRI, Rome & JIRCAS, Tsukuba, pp 8–20

  • Engelmann F (2004) Plant cryopreservation: progress and prospects. In Vitro Cell Dev Biol-Plant 40:427–433

    Article  Google Scholar 

  • Fábián A, Jager K, Darkó É, Barnabás B (2008) Cryopreservation of wheat (Triticum aestivum L.) egg cells by vitrification. Acta Physiol Plant 30:737–744

    Article  Google Scholar 

  • Gonzalez-Arnao MT, Panta A, Roca WM, Escobar RH, Engelmann F (2008) Development and large scale application of cryopreservation techniques for shoot and somatic embryo cultures of tropical crops. Plant Cell Tiss Organ Cult 92:1–13

    Article  Google Scholar 

  • Helliot B, Swennen R, Poumay Y, Frison E, Lepoivre P, Panis B (2003) Ultrastructural changes associated with cryopreservation of banana (Musa spp.) highly proliferating meristems. Plant Cell Rep 21:690–698

    PubMed  CAS  Google Scholar 

  • Kaviani B (2011) Conservation of plant genetic resources by cryopreservation. Afr J Crop Sci 5(6):778–800

    Google Scholar 

  • Khawnium T, Te-chato S (2011) Simple vitrification protocol for cryopreservation of oil palm using embryogenic culture. J Agr Technol 7(2):519–529

    Google Scholar 

  • Kohmura H, Sakai A, Chokyu S, Yakuwa T (1992) Cryopreservation of in vitro-cultured multiple bud clusters of asparagus [Asparagus officinalis L. cv. Hiroshimagreen (2np30)] by the techniques of vitrification. Plant Cell Rep 11:433–437

    Article  Google Scholar 

  • Mikuła A, Niedzielski M, Rybczynski JJ (2006) The use of TTC reduction assay for assessment of Gentiana spp. cell suspension viability after cryopreservation. Acta Physiol Plant 28(4):315–324

    Article  Google Scholar 

  • Mora-Urpí J, Weber JC, Clement CR (1997) Peach palm (Bactris gasipaes Kunth). Institute of Plant Genetics and Crop Plant Research and International Plant Genetic Resources Institute, Rome, p 81

    Google Scholar 

  • Morel G, Wetmore RH (1951) Tissue culture of monocotyledons. Am J Bot 38:138–140

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Niino T, Sakai A, Yakuwa H, Nojiri K (1992) Cryopreservation of in vitro-grown shoot tips of apple and pear by vitrification. Plant Cell Tiss Organ Cult 28:261–266

    Article  Google Scholar 

  • Nishizawa S, Sakai A, Amano Y, Matsuzawa T (1993) Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant-regeneration by vitrification. Plant Sci 91:67–73

    Article  CAS  Google Scholar 

  • Poobathy R, Hesam NA, Julkifle AL, Subramaniam S (2012) Vitrification and histological analyses of protocorm-like bodies of Vanda Kaseem’s Delight orchid. Aust J Crop Sci 6(2):219–224

    CAS  Google Scholar 

  • Reynolds ES (1963) The use of lead citrate at light pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–212

    Article  PubMed  CAS  Google Scholar 

  • Sen-Rong H, Ming-Hua Y (2012) A simple and efficient protocol for cryopreservation of embryogenic calli of the medicinal plant Anemarrhena asphodeloides Bunge by vitrification. Plant Cell Tiss Organ Cult 109:287–296

    Article  Google Scholar 

  • Sershen PB, Pammenter NW, Wesley-Smith J (2012a) The effects of various parameters during processing for cryopreservation on the ultrastructure and viability of recalcitrant zygotic embryos of Amaryllis belladonna. Protoplasma 249:155–169

    Article  PubMed  CAS  Google Scholar 

  • Sershen PB, Pammenter NW, Wesley-Smith J (2012b) Rate of dehydration, state of subcellular organization and nature of cryoprotection are critical factors contributing to the variable success of cryopreservation: studies on recalcitrant zygotic embryos of Haemanthus montanus. Protoplasma 249:171–186

    Article  PubMed  CAS  Google Scholar 

  • Schmidt ÉC, Pereira B, Pontes CLM, dos Santos R, Scherner F, Horta PA, Martins RP, Latini A, Maraschin M, Bouzon ZL (2012a) Alterations in architecture and metabolism induced by ultraviolet radiation-B in the carragenophyte Chondracanthus teedei (Rhodophyta, Gigartinales). Protoplasma 249:353–367

    Article  PubMed  CAS  Google Scholar 

  • Schmidt ÉC, Pereira B, Santos R, Gouveia C, Costa GB, Faria GSM, Scherner F, Horta PA, Paula MR, Latini A, Ramlov F, Maraschin M, Bouzon ZL (2012b) Responses of the macroalgae Hypnea musciformis after in vitro exposure to UV-B. Aquat Bot 100:8–17

    Article  CAS  Google Scholar 

  • Sokal RR, Rohlf FJ (1995) Biometry: the principles of statistics in biological research. Freeman, New York, p 887

    Google Scholar 

  • Steinmacher DA, Saldanha CW, Clement CR, Guerra MP (2007) Cryopreservation of peach palm zygotic embryos. CryoLetters 28:13–22

    PubMed  Google Scholar 

  • Steinmacher DA, Guerra MP, Saare-Surminski K, Lieberei R (2011) A temporary immersion system improves in vitro regeneration of peach palm through secondary somatic embryogenesis. Ann Bot. doi:10.1093/aob/mcr033

    PubMed  Google Scholar 

  • Suranthran P, Gantait S, Sinniah UR, Subramaniam S, Alwee SSRS, Roowi SH (2012) Effect of loading and vitrification solutions on survival of cryopreserved oil palm polyembryoids. Plant Growth Regul 66:101–109

    Article  CAS  Google Scholar 

  • Wang JH, Ge JG, Liu F, Huang CN (1998) Ultrastructural changes during cryopreservation of rice (Oryza sativa L.) embryogenic suspension cells by vitrification. CryoLetters 19:49–54

    Google Scholar 

  • Wesley-Smith J, Pammenter NW, Berjak P, Walters C (2001) The effects of two drying rates on the desiccation tolerance of recalcitrant jackfruit (Artocarpus heterophyllus Lamk.) seeds. Ann Bot 88:653–664

    Article  Google Scholar 

  • Wen B, Cai C, Wang R, Song S, Song J (2012) Cytological and physiological changes in recalcitrant Chinese fan palm (Livistona chinensis) embryos during cryopreservation. Protoplasma 249:323–335

    Article  PubMed  CAS  Google Scholar 

  • Yil JY, Sylvestre I, Colin M, Salma M, Lee SY, Kim HH, Park HJ, Engelmann F (2012) Improved cryopreservation using droplet-vitrification and histological changes associated with cryopreservation of madder (Rubia akane Nakai). Kor J Hort Sci Technol 30:79–84

    Google Scholar 

Download references

Acknowledgments

The authors thank the Coordination for the Improvement of Higher Education Personnel (CAPES) for fellowship to ASH and the National Council for Scientific and Technological Development (CNPq) for financial support in grants and fellowships to DAS and MPG.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Graduate Program in Plant Genetic Resources, Plant Developmental Physiology and Genetics Laboratory, Federal University of Santa Catarina, Rod. Admar Gonzaga, Km 3, Florianópolis, SC, 88034-001, Brazil

    Angelo Schuabb Heringer & Miguel Pedro Guerra

  2. Biosomatica Institute, Holambra, SP, 13825-000, Brazil

    Douglas André Steinmacher

  3. Post-Graduate Program in Cell Biology and Development, Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, 88049-900, CP 476, Florianópolis, SC, Brazil

    Éder Carlos Schmidt

  4. Central Laboratory of Electron Microscopy, Federal University of Santa Catarina, 88049-900, CP 476, Florianópolis, SC, Brazil

    Zenilda Laurita Bouzon

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  1. Angelo Schuabb Heringer
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  2. Douglas André Steinmacher
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  3. Éder Carlos Schmidt
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  4. Zenilda Laurita Bouzon
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  5. Miguel Pedro Guerra
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Correspondence to Miguel Pedro Guerra.

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Handling Editor: Friedrich W. Bentrup

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Heringer, A.S., Steinmacher, D.A., Schmidt, É.C. et al. Survival and ultrastructural features of peach palm (Bactris gasipaes, Kunth) somatic embryos submitted to cryopreservation through vitrification. Protoplasma 250, 1185–1193 (2013). https://doi.org/10.1007/s00709-013-0500-4

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  • DOI: https://doi.org/10.1007/s00709-013-0500-4

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