Abstract
The physicochemical properties and inherent ion content of the gelling agents used for the preparation of semi-solid substrates significantly affect the germination of tomato pollen in vitro. The addition of Ca, K and Mg to semi-solid, agar-based, substrates improved the germination of tomato pollen when the inherent Ca content of the agar was low, but was without effect or even inhibitory when the Ca level was high. However, Κ and/or Mg addition was beneficial irrespective of the agar source. When agarose replaced agar and K, Mg and Ca were added individually or in combination, pollen germination and pollen tube growth were affected differently by each ion but were optimal only in the presence of all three ions, reflecting the absence of these ions in agarose. An involvement of Na was also implicated since reduction of the inherently high Na content of agar by washing improved germination, with or without the addition of Κ, Mg and Ca. Since >3 mM Ca in the semi-solid substrate impairs tomato pollen germination, the gelling agent must be of high purity, which in the case of agar may entail washing, followed by the addition of K, Mg and Ca. The adoption of such a medium would permit the standardization of semi-solid substrates for in vitro tomato pollen germination studies.
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References
Anonymous (2001) Potassium and calcium in wine. Meterlab Applications – MTL008–2001–06B: Radiometer Analytical SA Lyon France 3 pp
Bergamini-Mulcahy G, Mulcahy DL (1988) The effect of supplemented media on the growth in vitro of bi- and trinucleate pollen. Plant Sci 55:213–216
Beruto M, Beruto D, Debergh P (1999a) Influence of agar on in vitro cultures: I. Physicochemical properties of agar and agar gelled media. In Vitro Cell Dev Biol Plant 35:86–93
Beruto M, Curir P, Debergh P (1999b) Influence of agar on in vitro cultures: II. Biological performance of Ranunculus on media solidified with three different␣agar brands. In Vitro Cell Dev Biol Plant 35:94–101
Beruto M, La Rosa C, Portogallo C (2001) Effects of agar impurities on in vitro propagation of Ranunculus asiaticus L. Acta Hortic 560:399–402
Brewbaker JL, Kwack BH (1963) The essential role of calcium in pollen germination and pollen tube growth. Am J Bot 50:859–865
Dafni A, Firmage D (2000) Pollen viability and longevity: practical, ecological and evolutionary implications. Plant Syst Evol 222:113–132
Debergh P (1983) Effects of agar brand and concentration on the tissue culture medium. Physiol Plant 59:270–276
Eaton AD, Clesceri LS, Greenberg AE (1995) Standard methods for the examination of water and wastewater, 19th edn. American Health Association, Washington
Feijo JA, Malhó R, Obermeyer G (1995) Ion dynamics and its possible role during in vitro pollen germination and tube growth. Protoplasma 187:155–167
Fernandez-Muňoz R, Gonzalez-Fernandez JJ, Cuartero J (1994) Methods for testing the fertility of tomato pollen formed at low temperature. J Hortic Sci 69:1083–1088
Gupta SK, Stewart JWB (1975) The extraction and determination of plant available boron in soil. Schweiz Landwirtsch Forsch 14:153–169
Heslop-Harrison J (1987) Pollen germination and pollen-tube growth. Int Rev Cytol 107:1–78
Heslop-Harrison JS (1992) Cytological techniques to assess pollen quality. In: Cresti M, Tiezzi A (eds) Sexual plant reproduction. Springer-Verlag, Berlin, pp 41–48
Hewitt FR, Hough T, O’Neill PO, Sasse JM, Williams EG, Rowan KS (1985) Effect of brassinolide and other growth regulators on the germination and growth of pollen tubes of Prunus avium using a multiple hanging-drop assay. Aust J Plant Physiol 12:201–211
Hoekstra FA (1992) Stress effects on the male gametophyte. In: Cresti M, Tiezzi A (eds) Sexual plant reproduction. Springer-Verlag, Berlin, pp 193–201
Holdaway-Clarke T, Hepler PK (2003) Control of pollen tube growth: role of ion gradients and fluxes. New Phytol 159:539–563
Karapanos IC, Passam HC, Olympios C, Akoumianakis K, Alexopoulos A (2004) Seasonal variation in the viability and production of pollen of greenhouse tomato (Lycopersicon esculentum Mill). Proc 21st Panhel Sci Conf Greek Soc Hort Sci 11A, 77–80
Karapanos IC, Fasseas K, Olympios C, Passam HC (2006) Factors affecting the efficacy of agar-based substrates for the study of tomato pollen germination. J Hortic Sci Biotech 81:631–638
Kordan HA (1981) Impatiens holstii pollen germination on non-nutrient agars. Plant Sci Lett 23:157–160
Kordan HA (1986) Boron in aqueous extracts of non-nutrient agars. Plant Soil 92:309–311
Kordan HA (1988) Inorganic ions present in commercial agars. Biochem Physiol Pflanz 183:355–359
Maisonneuve B, den Nijs APM (1984) In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth. Euphytica 33:833–840
McLeod KA (1975) The control of growth of tomato pollen. Ann Bot 39:591–596
Messerli MA, Smith PJS, Lewis RC, Robinson KR (2004) Chloride fluxes in lily pollen tubes: a critical reevaluation. Plant J 40:799–812
Nepi M, Franchi GG (2000) Cytochemistry of mature angiosperm pollen. Plant Syst Evol 222:45–62
Picton JM, Steer MW (1982) A model for the mechanism of tip extension in pollen tubes. J Theor Biol 98:15–20
Picton JM, Steer MW (1983) Evidence for the role of Ca2+ ions in tip extension in pollen tubes. Protoplasma 115:11–17
Scholten HJ, Pierik RLM (1998) Agar as a gelling agent: chemical and physical analysis. Plant Cell Rep. 17:230–235
Song J, Nada K, Tachibana S (1999) Ameliorative effect of polyamines on the high temperature inhibition of in vitro pollen germination in tomato (Lycopersicon esculentum Mill.). Sci Hortic 80:203–212
Stanley RG, Linskens HF (1974) Pollen: biology, biochemistry and management. Springer, New York
Steer MW, Steer JM (1989) Pollen tube tip growth. New Phytol 111:323–358
Taylor LP, Hepler PK (1997) Pollen germination and tube growth. Ann Rev Plant Physiol 48:461–491
Weisenseel MH, Jaffe LF (1976) The major growth current through lily pollen tubes enters as K+ and leaves as H+. Planta 133:1–7
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We thank Associate Professor K. Fasseas for provision of the optical microscope, Ms. A. Kefi and Mr. D. Papacharalambous for technical assistance.
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Karapanos, I.C., Akoumianakis, K., Olympios, C.M. et al. The influence of gelling agent purity and ion additions on in vitro tomato pollen germinability and pollen tube growth on semi-solid substrates. Plant Cell Tiss Organ Cult 87, 181–190 (2006). https://doi.org/10.1007/s11240-006-9152-9
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DOI: https://doi.org/10.1007/s11240-006-9152-9