Introduction: This study deals with the effects of extremely low doses or high dilutions of pharmacological compounds on in vitro pollen germination of kiwifruit (Actinidia deliciosa). Material and Methods: The biological efficacy of As2O3 at the 5th and 45th decimal dilution/succussion level (As 5x and As 45x) was tested on pollen previously stressed with As2O3 150 or 200 μM. The outcome variable was the pollen germination rate, as detected blind after 3 h 30 min by an Axioplan microscope. Results: A directionally consistent recovery of germination percentage was observed in both As 5x and As 45x after stressing with As2O3 150 μM. When pollen was stressed with As2O3 200 μM only as 45x induced a general and significant germination increase. Conclusions: Our results suggest that both treatments might partially remove the inhibitory effect caused by the stressor. Similar effects were observed on the same model using weak extremely low frequency magnetic fields (ELF-MFs) mediated through water. Although preliminary, the findings seem to indicate the in vitro pollen performance as adequate to study the effects of physicochemical subthreshold stimuli (extremely low doses or high dilutions of pharmacological compounds, weak ELF-MFs), mediated through water.

1.
Majewsky V, Arlt S, Shah D, Scherr C, Jäger T, Betti L, Trebbi G, Bonamin L, Klocke P, Baumgartner S: Use of homeopathic preparations in experimental studies with healthy plants. Homeopathy 2009;98:228-243.
2.
Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jäger T, Baumgartner S: Use of homeopathic preparations in phytopathological models and field trials: a critical review. Homeopathy 2009;98:244-266.
3.
Jäger T, Scherr C, Shah D, Majewsky V, Betti L, Trebbi G, Bonamin L, Simões-Wüst AP, Wolf U, Simon M, Heusser P, Baumgartner S: Use of homeopathic preparations in experimental studies with abiotically stressed plants. Homeopathy 2011;100:275-287.
4.
Endler P, Thieves K, Reich C, Matthiessen P, Bonamin L, Scherr C, Baumgartner S: Repetitions of fundamental research models for homeopathically prepared dilutions beyond 10(-23): a bibliometric study. Homeopathy 2010;99:25-36.
5.
Holandino C, Leal FD, de Oliveira Barcellos B, Campos MA, Oliveira R, da Veiga VF, Garcia S, Zacharias CR: Mechanical versus handmade succussions: a physical chemistry comparison; in Bonamin LV (ed): Signal and Images. London, Springer, 2008, pp 37-48.
6.
Shang A, Huwiler-Müntener K, Nartey L, Juni P, Dorig S, Sterne JA, Pewsner D, Egger M: Are the clinical effects of homeopathy placebo effects? Comparative study of placebo-controlled trials of homeopathy and allopathy. Lancet 2005;366:726-732.
7.
Betti L, Trebbi G, Nani D, Majewsky V, Scherr C, Jäger T, Baumgartner S: Models with plants, microorganism and viruses for basic research in homeopathy; in Bonamin LV (ed): Signal and Images. London, Springer, 2008, pp 97-111.
8.
Betti L, Trebbi G, Zurla M, Nani D, Peruzzi M, Brizzi M: A review of three simple plant models and corresponding statistical tools for basic research in homeopathy. ScientificWorldJournal 2010;10:2330-2347.
9.
Betti L, Trebbi G, Fregola F, Zurla M, Mesirca P, Brizzi M, Borghini F: Weak static and extremely low frequency magnetic fields affect in vitro pollen germination. ScientificWorldJournal 2011;11:875-890.
10.
Derksen J: Pollen tubes: a model system for plant growth. Bot Acta 1996;109:341-342.
11.
Taylor L P, Hepler PK: Pollen germination and tube growth. Annu Rev Plant Physiol Plant Mol Biol 1997;48:461-491.
12.
Li H, Lin Y, Heath RM, Zhu MX, Yang Z: Control of pollen tube tip growth by a Rop GTPase-dependent pathway that leads to tip-localized calcium influx. Plant Cell 1999;11:1731-1742.
13.
Feijò JA, Saihas J, Holdaway-Clarke T, Cordeiro MS, Kunkel JG, Helper PK: Cellular oscillations and the regulation of growth: the pollen tube paradigm. Bioessays 2001;23:86-94.
14.
Calzoni GL, Borghini F, Del Giudice E, Betti L, Dal Rio F, Migliori M, Trebbi G, Speranza A: Weak extremely high frequency microwaves affect pollen-tube emergence and growth in kiwifruit: pollen grain irradiation and water-mediated effects. J Altern Complement Med 2003;9:217-233.
15.
Holdaway-Clarke TL, Hepler PK: Control of pollen tube growth: role of ion gradients and fluxes. New Phytol 2003;159:539-563.
16.
Hepler PK: Calcium: a central regulator of plant growth and development. Plant Cell 2005;17:2142-2155.
17.
Brizzi M, Elia V, Trebbi G, Nani D, Peruzzi M, Betti L: The efficacy of ultramolecular aqueous dilutions on a wheat germination model as a function of heat and aging-time. Evid Based Complement Altern Med 2011;2011:696298.
18.
Bellavite P, Ortolani R, Conforti A: Immunology and homeopathy: 3 experimental studies on animal models. Evid Based Complement Altern Med 2006;3:171-186.
19.
Ozturk F, Duman F, Leblebici Z, Temizgul R: Arsenic accumulation and biological responses of water cress (Nasturtium officinale R. Br.) exposed to arsenite. Environ Exp Bot 2010;69:167-174.
20.
Dho S, Camusso W, Mucciarelli M, Fusconi A: Arsenate toxicity on the apices of Pisum saltivum L. seedling roots: effect on mitotic activity, chromatin integrity and microtubules. Environ Exp Bot 2010;69:17-23.
21.
Manomita P, Niladri B, Bulbul B, Archana S: Comparison of mercury, lead and arsenic with respect to genotoxic effects on plant systems and the development of genetic tolerance. Environ Exp Bot 2004;52:199-223.
22.
Li CX, Feng SL, Shao Y, Jiang LN, Lu XY, Hou XL: Effects of arsenic on seed germination and physiological activities of wheat seedlings. J Environ Sci 2007;19:725-732.
23.
Lin A, Zhang X, Zhu YG, Zhao FJ: Arsenate-induced toxicity: effects on antioxidative enzymes and DNA damage in Vicia faba. Environ Toxicol Chem 2008;27:413-419.
24.
Gunes A, Pilbeam DJ, Inal A: Effect of arsenic-phosphorous interaction on arsenic-induced oxidative stress in chickpea plants. Plant Soil 2009;314:211-220.
25.
Yi H, Wu L, Jiang L: Genotoxicity of arsenic evaluated by Allium-root micronucleus assay. Sci Total Environ 2007;383:232-236.
26.
Kristen U: Use of higher plants as screens for toxicity assessment. Toxicol In Vitro 1997;11:181-191.
27.
Baumgartner S, Heusser P, Thurneysen A: Methodological standards and problems in periclinical homeopathic potency research. Forsch Komplementarmed 1998;5:27-32.
28.
Stock-Schroer B, Albrecht H, Betti L, Endler PC, Linde K, Lüdtke R, Musial F, van Wijk R, Witt C, Baumgartner S: Reporting experiments in homeopathic basic research (REHBaR) - a detailed guideline for authors. Homeopathy 2009;98:287-298.
29.
Shah-Rossi D, Heusser P, Baumgartner S: Homeopathic treatment of Arabidopsis thaliana plants infected with Pseudomonas syringae. ScientificWorldJournal 2009;9:320-330.
30.
Yinnon TA, Yinnon CA: Electric dipole aggregates in very dilute polar liquids: theory and experimental evidence. IJMPB 2011;25:3707-3743.
31.
Calzoni GL, Speranza A, Bagni N: In vitro germination of apple pollens. Scientia Horticulturae 1979;10:49-55.
32.
Bomben C, Malossini C, Cioriani G, Testolin R: Long term storage of kiwifruit pollen. Hacta Hortic 1999;12:56-62.
33.
Spiegel MR, Schiller JJ, Srinivasan RA: Probability and Statistics. New York, McGraw-Hill, 2000, pp 408.
34.
Gosh S: Wetland macrophytes as toxic metal accumulators. Int J Environ Sci 2010;1:523-528.
35.
Singh HP, Batish DR, Kohli RK, Arora K: Arsenic-induced root growth inhibition in mung bean (Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation. Plant Growth Regul 2007;53:65-73.
36.
Shri M, Kumar S, Chkrabarty D, Trivedi PK, Mallick S, Misra P, Shukla D, Mishra S, Srivastava S, Tripathi RD, Tuli R: Effect of arsenic on growth, oxidative stress, and antioxidant system in rice seedlings. Ecotoxicol Environ Saf 2009;72:1102-1110.
37.
Xiong ZT, Peng YH: Response of pollen germination and tube growth to cadmium with special reference to low concentration exposure. Ecotoxicol Environ Saf 2001;48:51-55.
38.
Gur N, Topdemir A: Effects of heavy metals (Cd++, Cu++, Pb++, Hg++) on pollen germination and tube growth of quince (Cydonia oblonga M.) and plum (Prunus domestica L.). Fresenius Environ Bull 2005;14:36-39.
39.
Sawidis T: Effect of cadmium on pollen germination and tube growth in Lilium longiflorum and Nicotiana tabacum. Protoplasma 2008;233:95-106.
40.
Päivöke AEA, Simola LK: Arsenate toxicity to Pisum sativum: mineral nutrients, chlorophyll content, and phytase activity. Ecotox Environ Safe 2001;49;111-121.
41.
Smith SE, Christophersen HM, Pope S, Smith FA: Arsenic uptake and toxicity in plants: integrating mycorrhizal influences. Plant Soil 2010;327:1-21.
42.
Betti L, Lazzarato L, Trebbi G, Brizzi M, Calzoni GL, Borghini F, Nani D: Effects of homeopathic arsenic on tobacco plant resistance to tobacco mosaic virus. Theoretical suggestions about system variability, based on a large experimental data set. Homeopathy 2003;92:195-202.
43.
Brizzi M, Nani D, Peruzzi M, Betti L: Statistical analysis of the effect of high dilutions of arsenic in a large data set from a wheat germination model. Br Hom J 2000;89:63-67.
44.
Brizzi M, Lazzarato L, Nani D, Borghini F, Peruzzi M, Betti L: A biostatistical insight into the As(2) O(3) high dilution effects on the rate and variability of wheat seedling growth. Forsch Komplementarmed Klass Naturheilkd 2005;12:277-283.
45.
Marschollek B, Nelle M, Wolf M, Baumgartner S, Heusser P, Wolf U: Effects of exposure to physical factors on homeopathic preparations as determined by ultraviolet light spectroscopy. ScientificWorldJournal 2010;10:49-61.
46.
Anick DJ, Ives JA: The silica hypothesis for homeopathy: physical chemistry. Homeopathy 2007;96:189-195.
47.
Giuliani L, Grimaldi S, Lisi A, D'Emilia E, Bobkova N, Zhadin M: Action of combined magnetic fields on aqueous solution of glutamic acid: the further development of investigations. Biomagn Res Technol 2008;6:1.
48.
Lisi A, Ledda M, De Carlo F, Foletti A, Giuliani L, D'Emilia E, Grimaldi S: Calcium ion cyclotron resonance (ICR) transfers information to living systems: effects on human epithelial cell differentiation. Electromagn Biol Med 2008;27:230-240.
49.
Pang XF, Deng B: Investigation of changes in properties of water under the action of a magnetic field. Sci China Ser G Phys Mech Astron 2008;51:1621-1632.
50.
Chaplin M: Water structure and science. www.lsbu.ac.uk/water (accessed 03.07.2013).
51.
Foletti A, Ledda M, D'Emilia E, Grimaldi S, Lisi A: Experimental finding on the electromagnetic information transfer of specific molecular signals mediated through the aqueous system on two human cellular models. J Altern Complement Med 2012;18:258-261.
52.
Demangeat JL: NMR water proton relaxation in unheated and heated ultrahigh aqueous dilutions of histamine: evidence for an air-dependent supramolecular organization of water. J Mol Liq 2009;144:32-39.
53.
Elia V, Napoli E, Niccoli M: Thermodynamic parameters for the binding process of the OH- ion with the dissipative structures. Calorimetric and conductometric titrations. J Therm Anal Calorim 2010;102:1111-1118.
54.
Miranda AR, Vannucci A, Pontuschka WM: Impedance spectroscopy of water in comparison with high dilutions of lithium chloride. Mater Res Innov 2011;15:302-309.
55.
Betti L, Elia V, Napoli E, Trebbi G, Zurla M, Nani D, Peruzzi M, Brizzi M: Biological effects and physico-chemical properties of extremely diluted aqueous solutions as a function of aging-time. Frontiers in Life Science 2012;5:3-4, 117-126.
56.
Czerlinski G, Ypma T: Domains of water molecules provide mechanisms of potentization in homeopathy.Water 2010;2:1-13.
You do not currently have access to this content.