| Reprinted from: International Journal of High Dilution
Research, v.7, issue 22, p.3034, March 2008
-with permission from Prof. Carlos Moacir Bonato Universidade Estadual
de Maringá (UEM) Department of Biology, State University
of Maringá, Paraná, Brazil
ABSTRACT
The effects of high dilutions of Cymbopogon winterianus (citronella)
on the growth and
germination of Sida rhombifolia are analyzed; 5 homeopathic dilutions
(3cH, 6cH, 12cH, 24cH,
30cH) and a control (water) were used, with 5 repetitions. Variables
analyzed were the primary
growth of the root system, length of the shoot, fresh mass total,
germination percentage and
germination speed index. All dilutions stimulated the primary growth
of the root. Dilutions 3cH,
6cH, 12cH and cH stimulated the growth of the aerial parts and 24cH
inhibited it. Dilutions 6cH,
12cH, 24cH and 30cH stimulated a larger production of fresh mass
than 3cH. Dilution 12cH
resulted in the largest germination percentage while 24cH in the
lowest. Dilution 12cH resulted
in the highest score in the germination speed index, while 3cH and
24cH in the lowest.
Key words: Homeopathic dilutions; Plants; Germination; Growth;
Cymbopogon winterianus; Sida rhombifolia
Introduction
As it is known, the formulation of the homeopathic therapeutic
system was based on experimentation and observation of the effects
of high dilutions of substances on healthy human beings. Experiments
on plants developed in Europe, India, Mexico and Brazil have shown
that high dilutions also elicit effects in plants.(1) Experimental
research on healthy plants show the potential of high dilutions
to influence the system and modify their dynamic patterns of behavior,
manifested in an increase of defensive substances, photosynthesis,
detoxification of aluminum and copper and alterations in the metabolism
of plants.(2,18) These results indicate that homeopathic preparations
may be useful from an agroecological perspective, as they favor
sustainability and autonomy in the rural environment as well as
the production of healthy food and environmental quality. Sida rhombifolia
L. is an invading seed (space)propagated species common in the southern
region of Brazil. It develops in yearly and perennial cultures,
orchards, gardens, pastureland and uncultivated plots; it competes
highly with agriculture due to the development of deep root systems
which may reach 50 cm deep with many secondary roots.(19,20) Cymbopogon winterianus Jowitt, hailing from India and popularly known as citronella,
is widespread in Brazil. It is a perennial herb which forms 1meter
high compact and strong clumps. Citronella has economical importance
in the production of essential oils employed in the aromatic industry
and as an insect repellant; furthermore, it has antimicrobial and
acaricide properties. (21)More than 80 substances are comprised,
from which citronellal (40%), citronellol, geraniol, limonene and
esters have particular importance.(22,24) According to Einhelling,(25)
alellochemicals may interfere in the metabolism of plants in several
aspects, including growth, photosynthesis, enzymatic activity, protein
synthsis, membrane permeability and transport. The present study
aimed to verify the response of seedlings of S. rhombifolia to homeopathic
dilutions of C. winterianus.
Materials and methods
The study was conducted at the Homeopathy and
Vegetation Physiology laboratory of the Department
of Biology of the State University of Maringa.
The initial tincture was prepared from leaves of
adult C. winterianus collected in the morning from
the botanic garden of the University in a proportion
1:10(p/v), 1 g leaves/10 g cereal alcohol 70%. After
15days
maceration, the solution was filtered and
the tincture was preserved in an amber flask.
Homeopathic dilutions were prepared from this
tincture following the Brazilian Homeopathic
Pharmacopoeia (FHBII).(26) The first dilution (1cH)
was prepared adding 200 µl (0.2 mL) of tincture to
19.8 mL (1/100) distilled water with posterior
succussion 100 times by mechanical arm dynamizer
(Model Denise 50). Dilutions prepared were 3cH,
6cH, 12cH, 24cH and 30cH. The flasks were cleaned
and sterilized according to FHBII or cleaned with
alcohol 70%, boiled for 1 hour and dried on stove at
140ºC for 1 hour.
Dormancy of S. rhombifolia seeds was interrupted
through mechanical (lime) and thermal scarification
by immersing the seeds in water at 100ºC for 2
minutes. 20 seeds were distributed in Petri dishes
(total=600 seeds) covered by Whatman#1 filter
paper imbibed with 5mL of different homeopathic
dilutions (3cH, 6cH, 12cH, 24cH and 30cH) and
water (control) and distributed homogeneously in
benches with light at room temperature for 7 days.
Variables essayed were: length of primary root and
length of shoot by a graded ruler; increase of fresh
biomass by analytic scale; percentage of germination
and germination speed index (GSI). Only seeds with
2.0 mmlong
root protrusion were taken into
account. Maguire’s equation (27) was used to
calculate GSI: GSI = G1/N1 + G2/N2 + G3/N3 + ...
Gn/Nn, where G1, G2, G3, ... Gn = number of seeds
germinated on the day of observation and N1, N2, N3,
... Nn = number of days after seed planting.
The experiment design was totally randomized and
included 5 repetitions. Data were analyzed by
ANOVA and discriminated mean compared by ScottKnott
test at 5% probability (SAEG 5.0).
Results
S. rhombifolia responded to all dilutions regarding the length
of primary root (Figure 1).
Figure 1. Effect of homeopathic preparation C. winterianus on the length of primary root (LPR) of S. rhombifolia
seedlings with 5 solutions. (Mean followed by same letters does
not differ statistically in Scott-Knott test at 5% probability).
The increase in the length of primary root, which significantly
differed from the control, shows the healthy state of the seedlings.
Several experiments using homeopathic dilutions in seed germination
and plant growth have also been reported.(16,2832) Following Lisboa
et al. (33) it may be suggested that the information carried by
homeopathic dilutions had an influence on the seedlings of S. rhombifolia
physically expressed as an increase in the length of the primary
root.
The length of shoots increased with dilutions 3cH,
6cH, 12cH and 30cH by comparison to the control.
Results were similar to those in the root system
(Figure 2). The effect of dilution 24cH was not
significantly different from the control’s. These
results are consistent with the notion of oscillation
as a characteristic of high dilutions.(34)
Figure 2 – Effect of homeopathic preparation
C. winterianus on the shoot length (SL) of S. rhombifolia seedlings
treated with 5 solutions. (Mean followed by the same letters does
not significantly differ by Scott-Knott test at 5% probability).
All dilutions but 3cH increased the production of
fresh biomass regarding the control. (Figure 3)
Although the mechanism(s) of action of high
dilutions on plants are still not known, results
suggest that somehow dilutions 6cH, 12cH, 24cH
and 30cH increased the metabolic efficiency of the
seedlings.
Figure 3 – Effect of homeopathic preparation
C. winterianus on the increase of fresh biomass (FB) of S. rhombifolia
seedlings treated with 5 solutions (Mean followed by the same letters
does not differ statistically by Scott-Knott test at 5% probability).
Germination percentage (GER) was affected by dilutions of C. winterianus.
Dilutions 6cH, 12cH and 30cH had a higher effect than 3cH and 24cH
(Figure 4).

Figure 4 – Effect of homeopathic preparation
C. winterianus on the percentage of germination (GER) of S. rhombifolia
seeds treated with 5 solutions. (Mean followed by the same letters
does not differ statistically by Scott-Knott test at 5% probability).

Other studies have shown that citronella in
ponderal doses inhibit the germination of seeds,
Craveiro and Mattos have suggested that the reason
is its monoterpene contents. Our study, conducted
with high dilutions of citronella obtained the
opposite results. It may be suggested that dilutions
6cH, 12cH and 30cH activated some enzymes
involved in endosperm store degradation and
transport to the embryo during germination.(35)
Experimental results indicate that GSI was affected
by dilutions of C. winterianus. Dilutions 6cH, 12cH
and 30cH were associated to a higher GSI, while
dilutions 3cH and 24cH were not different from the
control. (Figure 5) It may be thought that C. winterianus had an effect on the metabolism of the
seeds of S. rhombifolia, intensifying enzyme
reactions, the transport of metabolites, digestion of
proteins, carbohydrates and lipids of the spare
tissue for cellular elongation retaking and
consequently of the emergence of radicles.
Figure 5 – Effect of homeopathic preparation
C. winterianus on germination speed index (GSI) in S. rhombifolia
seeds, with 5 solutions. (Mean followed by the same letters does
not differ statistically by Scott-Knott test at 5% probability).

Conclusions
All 5 dilutions of C. winterianus increased the
primary root of S. rhombifolia seedlings; all
dilutions but 24cH elicited an increase in the aerial
region. Seedlings exhibited a higher level of fresh
biomass production under the influence of all
dilutions but 3cH. Regarding germination
percentage, dilutions 6cH, 12cH and 30cH elicited a
higher score, which was not observed with dilutions
3cH and 24cH. Finally, the germination speed index
was higher with dilutions 6cH, 12cH and 30cH,
which was not the case of dilutions 3cH and 24cH.
These results indicate that homeopathic dilutions of
C. winterianus have an effect on the germination
and growth of S. rhombifolia. Moreover, that these
effects have an oscillatory pattern, which is
consistent with current conceptions of the
particularities of the biological effects of high
dilutions.
No mechanism of action may be still suggested for the biological
effects of high dilutions in general and on plants, in particular.
The results of this study suggest that several metabolic features
of plants might be involved. Further studies are needed to confirm
the experimental data, to assess the possible significance of their
application in agronomy and to postulate mechanism(s) of action
of high dilutions on plants.
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Support: none declared
Conflict of interest: none declared
Received: 13 February 2008; Revised: 19 March 2008; Published:
31 March 2008
correspondence author:
Prof. Carlos Moacir Bonato
Universidade Estadual de Maringá (UEM)
Depto de Biologia - Bloco H-78, Sala 20 Av. Colombo, 5790 87020-900
- Maringá – Pr Tel. 44 3261-4675 UEM 44 3261-3789 LAB/UEM
44 3026-5208 RES 44 9922-0960 CEL |