Acknowledgments
We thank diverse researchers and technicians from RBG Kew and
the MSB that have supported and revised the development of these
protocols, particularly Nicola Mills and Kirstine Manger. We also
thank Hongyin Chen (Chinese Academy of Agricultural Sciences)
and Daniela Impe (Leibniz Institute of Plant Genetics and Crop
Plant Research) for their work on pollen desiccation tolerance and
cryopreservation for some species. Royal Botanic Gardens, Kew
receives grant-in-aid from Defra.
References
1. Li DZ, Pritchard HW (2009) The science and
economics of ex situ plant conservation.
Trends Plant Sci 14:614–621
2. ENSCONET (2009) ENSCONET curation
protocols & recommendations. Royal Botanic
Gardens, Kew, UK. http://ensconetmaichgr/
PDF/Curation_protocol_Englishpdf.
Accessed 28 July 2019
3. FAO (2014) Genebank standards for plant
genetic resources for food and agriculture,
Rev. FAO, Rome
4. Royal Botanic Gardens Kew (2014) Millennium Seed Bank’s technical information sheets.
Board of Trustees of the Royal Botanic Gardens, Kew. http://brahmsonline.kew.org/
msbp/Training/Resources. Accessed 28 July
2019
5. Ballesteros D, Pence VC (2018) Fern conservation: spore, gametophyte, and sporophyte ex
situ storage, in vitro culture, and cryopreservation. In: Fernandez H (ed) Current advances in
fern research. Springer, New York, pp 227–249
6. Kumar A, Ferna ´ndez H, Revilla-Bahillo A
(2010) Working with ferns. Issues and applications. Springer, New York
7. Ballesteros D, Estrelles E, Walters C, Ibars AM
(2012) Effects of temperature and desiccation
on ex situ conservation of non-green fern
spores. Am J Bot 99:1–9
8. Ballesteros D, Hill LM, Walters C (2017) Variation of desiccation tolerance and longevity in
fern spores. J Plant Physiol 211:53–62
9. Ballesteros D, Hill LM, Lynch RT, Pritchard
HW, Walters C (2019) Longevity of preserved
germplasm: the temperature dependency of
aging reactions in glassy matrices of dried fern
spores. Plant Cell Physiol 60:376–392
10. Li Y, Shi L (2014) Effect of desiccation level
and storage temperature on green spore viability of Osmunda japonica. Cryobiology
68:446–450
11. Ballesteros D (2010) Conservation of fern
spores. In: Kumar A, Ferna ´ndez H, RevillaBahillo A (eds) Working with ferns. Issues and
applications. Springer, New York, pp 165–172
12. Pence VC (2000) Survival of chlorophyllous
and non-chlorophyllous fern spores through
exposure to liquid nitrogen. Am Fern J
90:119–126
13. Ballesteros D, Estrelles E, Walters C, Ibars AM
(2011) Effect of storage temperature on green
spore longevity for the ferns Equisetum ramosissimum and Osmunda regalis. CryoLetters
32:89–98
14. Mikula A, Tomiczak K, Makowski D,
Niedzielski M, Rybczyn ´ ski JJ (2015) The effect
of moisture content and temperature on spore
aging in Osmunda regalis. Acta Physiol Plant
37:229
15. Ganeshan S, Rajasekharan PE (2000) Current
status of pollen cryopreservation research: relevance
to
tropical
horticulture.
In:
Engelmann F, Takagi H (eds) Cryopreservation of tropical plant germplasm - current
research progress and applications. JIRCAS/
IPGRI Publication, Tsukuba/Rome, pp
360–365
16. Benson EE (2008) Cryopreservation of phytodiversity: a critical appraisal of theory & practice. Crit Rev Plant Sci 27:141–219
17. Towill LE, Walters C (2000) Cryopreservation
of pollen. In: Engelmann F, Takagi H (eds)
Cryopreservation of tropical plant germplasm
- current research progress and applications.
JIRCAS/IPGRI Publication, Tsukuba/Rome,
pp 115–129
18. Wang MR, Chen L, Teixeira da Silva JA, Volk
GM, Wang QC (2018) Cryobiotechnology of
apple (Malus spp.): development, progress and
future prospects. Plant Cell Rep 37:689–709
19. Engelmann F (2011) Use of biotechnologies
for the conservation of plant biodiversity. In
Vitro Cell Dev Biol Plant 47:5–16
Fern Spores and Pollen Cryopreservation
635
We thank diverse researchers and technicians from RBG Kew and
the MSB that have supported and revised the development of these
protocols, particularly Nicola Mills and Kirstine Manger. We also
thank Hongyin Chen (Chinese Academy of Agricultural Sciences)
and Daniela Impe (Leibniz Institute of Plant Genetics and Crop
Plant Research) for their work on pollen desiccation tolerance and
cryopreservation for some species. Royal Botanic Gardens, Kew
receives grant-in-aid from Defra.
References
1. Li DZ, Pritchard HW (2009) The science and
economics of ex situ plant conservation.
Trends Plant Sci 14:614–621
2. ENSCONET (2009) ENSCONET curation
protocols & recommendations. Royal Botanic
Gardens, Kew, UK. http://ensconetmaichgr/
PDF/Curation_protocol_Englishpdf.
Accessed 28 July 2019
3. FAO (2014) Genebank standards for plant
genetic resources for food and agriculture,
Rev. FAO, Rome
4. Royal Botanic Gardens Kew (2014) Millennium Seed Bank’s technical information sheets.
Board of Trustees of the Royal Botanic Gardens, Kew. http://brahmsonline.kew.org/
msbp/Training/Resources. Accessed 28 July
2019
5. Ballesteros D, Pence VC (2018) Fern conservation: spore, gametophyte, and sporophyte ex
situ storage, in vitro culture, and cryopreservation. In: Fernandez H (ed) Current advances in
fern research. Springer, New York, pp 227–249
6. Kumar A, Ferna ´ndez H, Revilla-Bahillo A
(2010) Working with ferns. Issues and applications. Springer, New York
7. Ballesteros D, Estrelles E, Walters C, Ibars AM
(2012) Effects of temperature and desiccation
on ex situ conservation of non-green fern
spores. Am J Bot 99:1–9
8. Ballesteros D, Hill LM, Walters C (2017) Variation of desiccation tolerance and longevity in
fern spores. J Plant Physiol 211:53–62
9. Ballesteros D, Hill LM, Lynch RT, Pritchard
HW, Walters C (2019) Longevity of preserved
germplasm: the temperature dependency of
aging reactions in glassy matrices of dried fern
spores. Plant Cell Physiol 60:376–392
10. Li Y, Shi L (2014) Effect of desiccation level
and storage temperature on green spore viability of Osmunda japonica. Cryobiology
68:446–450
11. Ballesteros D (2010) Conservation of fern
spores. In: Kumar A, Ferna ´ndez H, RevillaBahillo A (eds) Working with ferns. Issues and
applications. Springer, New York, pp 165–172
12. Pence VC (2000) Survival of chlorophyllous
and non-chlorophyllous fern spores through
exposure to liquid nitrogen. Am Fern J
90:119–126
13. Ballesteros D, Estrelles E, Walters C, Ibars AM
(2011) Effect of storage temperature on green
spore longevity for the ferns Equisetum ramosissimum and Osmunda regalis. CryoLetters
32:89–98
14. Mikula A, Tomiczak K, Makowski D,
Niedzielski M, Rybczyn ´ ski JJ (2015) The effect
of moisture content and temperature on spore
aging in Osmunda regalis. Acta Physiol Plant
37:229
15. Ganeshan S, Rajasekharan PE (2000) Current
status of pollen cryopreservation research: relevance
to
tropical
horticulture.
In:
Engelmann F, Takagi H (eds) Cryopreservation of tropical plant germplasm - current
research progress and applications. JIRCAS/
IPGRI Publication, Tsukuba/Rome, pp
360–365
16. Benson EE (2008) Cryopreservation of phytodiversity: a critical appraisal of theory & practice. Crit Rev Plant Sci 27:141–219
17. Towill LE, Walters C (2000) Cryopreservation
of pollen. In: Engelmann F, Takagi H (eds)
Cryopreservation of tropical plant germplasm
- current research progress and applications.
JIRCAS/IPGRI Publication, Tsukuba/Rome,
pp 115–129
18. Wang MR, Chen L, Teixeira da Silva JA, Volk
GM, Wang QC (2018) Cryobiotechnology of
apple (Malus spp.): development, progress and
future prospects. Plant Cell Rep 37:689–709
19. Engelmann F (2011) Use of biotechnologies
for the conservation of plant biodiversity. In
Vitro Cell Dev Biol Plant 47:5–16
Fern Spores and Pollen Cryopreservation
635