Register      Login
Australian Journal of Botany Australian Journal of Botany Society
Southern hemisphere botanical ecosystems
RESEARCH ARTICLE

Optimising storage and in vitro germination of Eucalyptus pollen

Tasmien N. Horsley A B C , Steven D. Johnson A and Terrence K. Stanger B
+ Author Affiliations
- Author Affiliations

A School of Biological and Conservation Sciences, University of KwaZulu-Natal, Scottsville, South Africa.

B Shaw Research Centre, Sappi Forests, Howick, South Africa.

C Corresponding author. Email: tasmien.horsley@sappi.com

Australian Journal of Botany 55(1) 83-89 https://doi.org/10.1071/BT05194
Submitted: 12 November 2005  Accepted: 31 July 2006   Published: 18 January 2007

Abstract

The best sucrose solution for maximum in vitro germination of Eucalyptus pollen was investigated in order to evaluate pollen germination rate as an indicator of pollen viability. In vitro germination of both freshly collected and 1-year-old pollen (stored at 4°C) of Eucalyptus grandis, E. smithii, E. nitens, E. dunnii and E. macarthurii was carried out in 0, 10, 20, 30, 40 and 50% (w/v) sucrose solutions, either with (0.15 mg L–1) or without boric acid. Similar trends were obtained for both fresh and 1-year-old pollen, with all species responding most favourably to 30% (w/v) sucrose and 0.15 mg L–1 boric acid. When an optimal in vitro germination medium had been established, the viabilities (%germination) of E. smithii, E. nitens and E. grandis pollen, stored at room (25°C), fridge (4°C), freezer (–10°C) and liquid nitrogen (–196°C) temperatures, were compared. For all tested species, germination declined as storage temperature increased, and by 8 months, the highest survival was obtained with cryostored pollen.


Acknowledgements

I thank Diana Madondo for her help with pollen collection and processing and Nicky Jones for the use of her tissue-culture laboratory for pollen storage and viability testing. Thanks go to Cathy Ford and Lorna Fisher for their assistance throughout the study.


References


Barnabas B , Kovacs G (1997) Storage of pollen. In ‘Pollen biotechnology for crop production and improvement’. (Eds KR Shivanna, VK Sawhney) (Cambridge University Press)

Benson EE (1994) Cryopreservation. In ‘Plant cell culture: a practical approach’. 2nd edn. (Eds RA Dixon, RA Gonzales) pp. 147–167. (IRL Press: Oxford, UK)

Boden RW (1958) Handling and storage of pollen in Eucalyptus breeding. Australian Forestry 22, 73–81. open url image1

Bonnet-Masimbert M, Baldet P, Paques L, Phillipe G (1998) From flowering to artificial pollination in larch for breeding and seed orchard production. Forestry Chronicles 74, 195–202. open url image1

Bowes SA (1990) Long-term storage of Narcissus anthers and pollen in liquid nitrogen. Euphytica 48, 275–278.
Crossref | GoogleScholarGoogle Scholar | open url image1

Brewbaker JL , Kwack BH (1964) The calcium ion and substances influencing pollen growth. In ‘Pollen physiology and fertilisation’. (Ed. HF Linskens) pp. 143–151. (North Holland: Amsterdam)

Buitink J, Walters C, Hoekstra F, Crane J (1998) Storage behaviour of Typha latifolia pollen at low water contents: interpretation on the basis of water activity and glass concepts. Physiologia Plantarum 103, 145–153.
Crossref | GoogleScholarGoogle Scholar | open url image1

Craddock JH, Reed SM, Schlarbaum SE, Sauve RJ (2000) Storage of flowering dogwood (Cornus florida L.) pollen. HortScience 35, 108–109. open url image1

Dafni A (1992) ‘Pollination ecology: a practical approach.’ (Oxford University Press: New York)

De Nettancourt D (1977) ‘Incompatibility in angiosperms.’ (Springer-Verlag: Berlin)

Dordas C, Brown PH (2000) Permeability of boric acid across lipid bilayers and factors affecting it. Journal of Membrane Biology 175, 95–105.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Dumont-BeBoux N, von Aderkas P (1997) In vitro pollen tube growth in Douglas-fir. Canadian Journal of Forestry Research 27, 674–678.
Crossref | GoogleScholarGoogle Scholar | open url image1

Eldridge KG , Davidson J , Harwood C , Van Wyk G (1994) ‘Eucalypt domestication and breeding.’ (Oxford University Press Inc.: New York)

Engelmann F (1997) Importance of desiccation for the cryopreservation of recalcitrant seed and vegetatively propagated species. Plant Genetic Resources Newsletter 112, 9–18. open url image1

Feijo JA, Malho R, Obermeyer G (1995) Ion dynamics and its possible role during in vitro pollen germination and tube growth. Protoplasma 187, 155–167.
Crossref | GoogleScholarGoogle Scholar | open url image1

Finkle JF , Zavala ME , Ulrich JM (1985) Cryoprotective compounds in viable freezing of plant tissues. In ‘Cryopreservation of plant cells and organs’. (Ed. KK Kartha) pp. 75–113. (CRC Press: Boca Raton, FL)

Frampton J (1997) Controlled-pollination of Fraser fir. Limbs & Needles 24, 10–15. open url image1

Ganeshan S (1986) Cryogenic preservation of papaya pollen. Scientia Horticulturae 28, 65–70.
Crossref | GoogleScholarGoogle Scholar | open url image1

Griffin AR, Ching KK, Johnson KW, Hand FC, Burgess IP (1982) Processing Eucalyptus pollen for use in controlled pollination. Silvae Genetica 31, 198–203. open url image1

Grout BWW (1995) Introduction to the in vitro preservation of plant cells, tissues and organs. In ‘Genetic preservation of plant cells in vitro’. (Ed. BWW Grout) pp. 1–20. (Springer-Verlag: Berlin)

Hanna WW , Towill LW (1995) Long term pollen storage. In ‘Plant breeding reviews’. (Ed. J Janic) pp. 197–207. (John Wiley and Sons: Chichester, UK)

Harbard JL, Griffin AR, Espejo J (1999) Mass controlled pollination of Eucalyptus globulus: a practical reality. Canadian Journal of Forest Research 29, 1457–1463.
Crossref | GoogleScholarGoogle Scholar | open url image1

Heslop-Harrison J, Heslop-Harrison Y (1985) Germination of stress-tolerant Eucalyptus pollen. Journal of Cell Science 73, 135–137.
PubMed |
open url image1

Janssen AWB, Hermsen JGT (1976) Estimating pollen fertility in Solanum species and haploids. Euphytica 25, 577–586.
Crossref | GoogleScholarGoogle Scholar | open url image1

Kearns CA , Inouye DW (1993) ‘Techniques for pollination biologists.’ (University Press of Colorado: Niwot Ridge, CO)

Lanteri S, Belletti P, Lotito S (1993) Storage of pollen of Norway spruce and different pine species. Silvae Genetica 42, 2–3. open url image1

Leduc N, Monnier M, Douglas GC (1990) Germination of trinucleated pollen: formulation of a new medium for Capsella bursapastoris. Sexual Plant Reproduction 3, 228–235.
Crossref | GoogleScholarGoogle Scholar | open url image1

Loguercio LL (2002) Pollen treatment in high osmotic potential: a simple tool for in vitro preservation and manipulation of viability in gametophytic populations. Brazilian Journal of Plant Physiology 14, 65–70.
Crossref | GoogleScholarGoogle Scholar | open url image1

Loomis WD, Durst RW (1991) Boron and cell walls. Current Topics in Plant Biochemistry and Physiology 10, 149–178. open url image1

Matthews FR, Bramlett DL (1986) Pollen quantity and viability affect seed yields from controlled pollinations of loblolly pine. Southern Journal of Applied Forestry 10, 78–80. open url image1

Obermeyer G, Blatt MR (1995) Electrical properties of intact pollen grains of Lilium longiflorum: characteristics of the non-germination grain. Journal of Experimental Botany 46, 803–813.
Crossref |
open url image1

Pammenter NW , Berjak P , Walters C (1999) The effect of drying rate on recalcitrant seeds: ‘lethal water contents’, causes of damage, and quantification of recalcitrance. In ‘Sixth international workshop on seeds’. (Eds M Black, KJ Bradford, JM Vazquez-Ramos) (CAB International: Wallingford, UK)

Potts BM, Marsden-Smedley JB (1989) In vitro germination of Eucalyptus pollen: response to variation in boric acid and sucrose. Australian Journal of Botany 37, 429–441.
Crossref | GoogleScholarGoogle Scholar | open url image1

Shivanna KR , Rangaswamy NS (1992) ‘Pollen biology: a laboratory manual.’ (Springer-Verlag: Berlin)

Stangoulis JCR, Reid RJ, Brown PH, Graham RD (2001) Kinetic analysis of boron transport in Chara. Planta 213, 142–146.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Vasil IK (1964) Effect of boron on pollen germination and pollen tube growth. In ‘Pollen physiology and fertilisation’. (Ed. HF Linskens) pp. 107–119. (North Holland: Amsterdam)

Wang Q, Lu L, Wu X, Li Y, Lin J (2003) Boron influences pollen germination and pollen tube growth in Picea meyeri. Tree Physiology 23, 345–351.
PubMed |
open url image1

Watt MP, Mycock DJ, Blakeway FC, Berjak P (2000) Applications of in vitro methods to Eucalyptus germplasm conservation. Southern African Forestry Journal 187, 3–10. open url image1