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Pollen Cryopreservation of Coniferous Serbian Spruce (Picea omorika/Panč./Purkyne) and Deciduous Pedunculate Oak (Quercus robur L.) Species

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Pollen Cryopreservation Protocols

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Abstract

One of the alternative strategies to conserve and preserve the genetic variation of many plant species is pollen cryopreservation. The aim of this study was to develop protocols for preserving pollen from coniferous Serbian spruce (Picea omorika/Panč./Purkyne) and deciduous pedunculate oak (Quercus robur L.) species. Although these species have different forms (coniferous vs. deciduous), both of them are monoecious and anemophilous. Samples from different localities over a few successive years were collected and stored at four temperatures (+23 °C, +5 °C, −15 °C, and −20 °C). Pollen viability analysis methods include protocols for collecting fresh pollen, storing it under controlled temperatures and light conditions, and microscopic analysis of pollen (viability %, germination %, germination energy-pollen tube length μm). Experiments to analyze the viability of stored pollen were set up every month during the first year of storage, every 3 months during the second year of storage, and once a year during the third and subsequent years of storage. The pollen vitality was observed by staining test (TTC-2,3,5-triphenyl tetrazolium chloride). Pollen viability was evaluated by pollen germination percentage and pollen tube length on six sucrose medium (0%, 5%, 10%, 15%, 20%, 25%) for fresh pollen and periodically for pollen after cryopreservation.

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Acknowledgments

This paper was financed by the Ministry of Education and Science of the Republic of Serbia (contract number 451-03-68/2020-14/200007). Gratitude: to the technical associate Miloš Bokorov, Faculty of Sciences, Department of Biology and Ecology, University Center for Electronic Microscopy, Novi Sad (UCEM-NS), who took Figs. 2 and 3. The other attached photos were taken by the author of the paper Dr. B. Batos, Institute for Forestry, Belgrade. The authors wish to thank the college Dr. Darka Šešlija Jovanović and English language editor, native speaker, and teacher of English Mrs. Esther Grace Helajzen for proofreading and revising text correction.

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Authors and Affiliations

  1. Department of Genetics, Institute of Forestry, Belgrade, Serbia

    Branislava Batos

  2. Department of Evolutionary Biology, Institute for Biological Research “SinišaStanković”–National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia

    Danijela Miljković

Authors
  1. Branislava Batos
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  2. Danijela Miljković
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Corresponding author

Correspondence to Danijela Miljković .

Editor information

Editors and Affiliations

  1. Division of Plant Genetic Resources, ICAR-Indian Institute of Horticultural Research, Bangalore, Karnataka, India

    P.E. Rajasekharan

  2. Division of Flower and Medicinal Crops, ICAR-Indian Institute of Horticultural Research, Bangalore, Karnataka, India

    M.R. Rohini

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Batos, B., Miljković, D. (2023). Pollen Cryopreservation of Coniferous Serbian Spruce (Picea omorika/Panč./Purkyne) and Deciduous Pedunculate Oak (Quercus robur L.) Species. In: Rajasekharan, P., Rohini, M. (eds) Pollen Cryopreservation Protocols . Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2843-0_53

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  • DOI: https://doi.org/10.1007/978-1-0716-2843-0_53

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2842-3

  • Online ISBN: 978-1-0716-2843-0

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