Abstract
Cryopreservation of pollen grains is an efficient technique to overcome asynchronous flowering and to support actions for genetic improvement and conservation of important alleles. It can be used both by germplasm curators and plant breeders. In the case of Bromeliaceae, a family with wide diversity but also high vulnerability, the form of conservation can be crucial to prevent the increasing problem of genetic erosion. This chapter describes a method of cryopreservation of pollen grains of different Bromeliaceae species, including pineapple, after dehydration with silica and subsequent immersion in liquid nitrogen. The efficiency of this protocol has been demonstrated by the high pollen viability percentage and production of seeds after in vivo pollination with cryopreserved grains. The protocol can be used for cryopreserving pollen of many species of bromeliads and is easy to perform.
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References
Souza EH, Souza FVD, Costa MAPC et al (2012) Genetic variation of the Ananas genus with ornamental potential. Genet Resour Crop Evol 59:1357–1376. https://doi.org/10.1007/s10722-011-9763-9
Souza EH, Souza FVD, Rossi ML et al (2015) Viability, storage and ultrastructure analysis of Aechmea bicolor (Bromeliaceae) pollen grains, an endemic species to the Atlantic forest. Euphytica 204:13–28. https://doi.org/10.1007/s10681-014-1273-3
Silva RL, Souza EH, Vieira LJ et al (2017) Cryopreservation of pollen of wild pineapple accessions. Sci Hortic 219:326–334. https://doi.org/10.1016/j.scienta.2017.03.022
Grout BWW, Roberts AV (1995) Storage of free pollen, pollen embryos and the zygotic embryos of seed by cryopreservation and freeze drying. In: Grout BWW (ed) Genetic preservation of plant cells in vitro. Springer-Verlag, Berlin, pp 63–74. https://doi.org/10.1007/978-3-642-78661-7_5
Tai PYP, Miller JD (2002) In vivo viability assay of sugarcane pollen stored at ultra low temperature following preservation treatments. J Am Soc Sugarcane Technol 22:135–136
Vendrame WA, Carvalho VS, Dias JMM, Maguire I (2008) Pollination of dendrobium hybrids using cryopreserved pollen. Hortscience 43:264–267
Parton E, Deroose R, De Proft MP (1998) Cryostorage of Aechmea fasciata pollen. CryoLetters 19:355–360
Parton E, Vervaeke R, Delen BR et al (2002) Viability and storage of bromeliad pollen. Euphytica 125:155–161. https://doi.org/10.1023/A:1015884019619
Towill LE (1985) Low temperature and freeze-vacuum-drying preservation of pollen. In: Kartha KK (ed) Cryopreservation of plant cells and organs. CRC Press, Boca Raton, pp 171–198
Akihama T, Omura M, Kosaki I (1979) Long-term of fruit tree pollen and its application in breeding. Trop Agr Res 13:238–241
Soares TL, Silva SO, Costa MAPC et al (2008) In vitro germination and viability of pollen grains of banana diploids. Crop Breed Appl Biot 8:111–118. https://doi.org/10.12702/1984-7033.v08n02a03
Yates IE, Sparks D (1989) Hydration and temperature influence in vitro germination of pecan pollen. J Am Soc Hortic Sci 114:599–605
Taylor LP, Hepler PK (1997) Pollen germination and tube growth. Annu Rev Plant Physiol Plant Mol Biol 48:461–491. https://doi.org/10.1146/annurev.arplant.48.1.461
Ganeshan S, Rajasekharan PE, Shashikumar S, Decruze W (2008) Cryopreservation of pollen. In: Reed BM (ed) Plant cryopreservation: a practical guide. Springer, New York, pp 281–332. https://doi.org/10.1007/978-0-387-72276-4_17
Benson EE (2008) Cryopreservation theory. In: Reed BM (ed) Plant cryopreservation. A practical guide. Springer, New York, pp 15–32. https://doi.org/10.1007/978-0-387-72276-4_2
Scorza R, Sgerman WB (1995) Peaches. In: Janik J, Morre JN (eds) Fruit breeding. John & Sons, New York, pp 325–440
Brewbaker JL, Kwack BH (1963) The essential role of calcium ion in pollen germination and pollen tube growth. Am J Bot 50:859–865
Souza EH (2013) Reprodução e hibridação interespecífica e intergenérica em bromeliáceas com potencial ornamental. Tesis (Doctoral) University of São Paulo, Piracicaba, São Paulo. p. 256
Souza EH, Souza FVD, Rossi, ML, Packer RM, Cruz-Barros MAV, Martinelli AP (2017) Pollen morphology and viability in Bromeliaceae. Ann Braz Acad Sci 89:3067–3082. http://dx.doi.org/10.1590/0001-3765201720170450
Acknowledgments
The authors acknowledge Fundação de Amparo a Pesquisa do Estado da Bahia (FAPESB), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)/EMBRAPA program, and Embrapa Mandioca e Fruticultura for financial support.
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Souza, F.V.D., de Souza, E.H., da Silva, R.L. (2018). Cryopreservation of Pollen Grains of Pineapple and Other Bromeliads. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 1815. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8594-4_19
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DOI: https://doi.org/10.1007/978-1-4939-8594-4_19
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