SYNERGISM OF ACTION BETWEEN INDOLOACETIC ACID (IAA) AND HIGH DILUTED SOLUTIONS OF CaCO3 IN 5°, 15°, 30° CH ON THE GROWTH IN LENGTHENING OF COLEOPTILE OF OATS
BORNORONI CORRADO, MD
Director of "Istituto Superiore di Medicina Olistica e di Ecologia"
Università degli Studi di Urbino
61029 Urbino (PS) - Italy
Abstract
This research work was conducted to assess the effect of high diluited solutions of CaCO3 on the growth in lengthening of coleoptile of oats stimulate by the IAA 100 mM (indoloacetic acid 10-4).
The specimens pre-trated 10 hrs before with a homoeopathic solutions wich obtained statistically valid increase, when compared with the percentage growth of the specimens with just IAA, where those of 5° CH.
KEY WORDS: CaCO3 5°-15°-30° CH with IAA. Homoeopathic synergism.
Introduction
The biological regulation which allow every unicellular organism, like every pluricellular, to coordinate the various metabolic activities of the cell (nutrition, growth, differentiation, reproduction) is strictly subordinate to a system articulated in 3 phases: stimulus-reception-response.
The nature of the stimulus can be physical (thermoinduction or photo-induction of morphogenetic process) or chemical. The nature of the reception is essentially less variable since bind fast to the protein structure and to the informational character, wich create specific conditionings:
a - recognition of the part of small molecules,
b - modification of its own configuration,
c - adaptation of its own activity on the basis of the structural modifications undergone.1
Characteristics of the hormones of plants
One fundamental characteristic of plant hormones is the almost total absence of specific production sites, both at the tissue and at the inter-cellular levels. Only the phototropic response of the coleoptile cereals can be ascribed to an endogenous site (place) of the auxina localized in the upper part (apex) and responsible for the growth of coleoptile in oats or wheat. The characteristics of plant hormones can be synthesized thus:
1. The action of each hormone is multiple in that it is manifested both in growth phenomena by moltiplication or by extension and in phenomena of differentiation.
2. The evaluation of the biological hormonal activity in made evident through its specific action of particular tissues and on different stages of development.
3. Every ormonal response is variable from a qualitative and a quantitative point of view, in relation to 2 parameters:
3.1. the levels of concentration of the hormone;
3.2. the variations in the tissue on which the hormone acts.
Consequently, many of the physiological process of the plants are under the control not of one single hormone, but of a poly-hormonal system.
From this one can deduce the difficulties in studying plant hormones, also because the reception-sites (places) of the hormones, the sites of hormonal biosynthesis and the degradation paths are still unknown.2
The physiology of auxine
As previously said, each functional manifestation cannot be attributed to the esclusive action of one hormone, since, using gascromatography (GLC) and high pressure liquid cromatography (HPLC), in conjunction with mass spectrometry (MS), in various vegetable tissues are found different classes of hormones capable of influencing the activities of the tissues themselves.
The auxines, of which the indoloacetic acid (IAA) is considered the most important from a physiological point of view, appear to be involved in many physiological phenomena and in limited functionally specific actions, among which:
a) cellular extension;
b) cellular division;
c) some processes of differentiation, including its role in root growth.
The optimum concentration in which the most incisive action of IAA on the tissues can be seen, oscillates betwen the values of 10-4 and 10-6 M, but its effect on the lengthening of roots grown "in vitro" has also been noted with doses of 10-13 M.3
The synthesis of IAA has been demonstrated in various tissues, particolarly young tissues with meristematic activity, such as the "apici caulinari", the gemmae, the germinating seeds, the flowers and the fruit in the initial phase of growth.4
Experimental
Research protocol
CaCO3, in doses of 1/1010, 1/1030, 1/1060 was chosen, since it could influenze neo-differentiation, acting principally as a cellular stimulant through a better reception of the calcium ion from the cultivation ground.
From the relevant literature, Kirkby et al. (1984), note that the mouvement of calcium in growth tissues takes place in the direction of the new cells in formation, and it is presumed that this migration is induced and stimolate by IAA.5
The obiective of this research project is drawn from these premises: to investigate and evaluate the effect of different dynamized dilutions of CaCO3, and to check whether or not an effect exists, even with such infinitesimally small doses, that exceed significantly the number of Avogadro, on the growth in length of the coleoptile of oats induced by IAA (10-4 M).
Hypothesis to verify
The research project was established, on the basis of the specific nature of hormonal action, to evaluate the possible synergism of action of different dynamized diluitions of CaCO3 on the growth of segments of coleoptile of oats, induced by concentration of IAA of 10-4 M.
Materials and methods
Small oat plants, grown in the dark until the coleoptile reaches a lenght of 2-3 cm, were used in this experiment. 10 segments of standard lenght (about 3-5 mm) where isolated from the sub-apex area, and placed in the solutions to be examined in the dark.6
1. In this way, the different growth responses in the segments of oats were investigated considering this treatment with IAA 100 mM (10-4 M), pretreated with CaCO3 dynamized in 5°CH, 15°CH, 30°CH , and with the respective dilutions of CaCO3, first heated to a temperature of 60° and then colded. The lengthening induced by the various solutions under examination, compared with the control specimen kept in plain water and dynamized, was taken as the measure of auxinic activity (IAA) both taken alone and with the various dilutions of CaCO3.
2. Further, the growth responses of the oats treated with IAA 100 mM (10-4 M), and of those treated only with the addition of CaCO3 dynamized to 5°CH, 15°CH and 30°CH, were compared.
EXPERIMENTAL PROTOCOL OF POINT 1
Graph n° 1
|
Control |
Dynamized control |
|
Treated with IAA |
Treated with IAA dynamized |
Graph n° 2
|
Pretreat. with CaCO3 5° CH dyn.(1) |
Pretreat. with CaCO3 5° CH.(2) |
|
Pretreat. with CaCO3 15° CH dyn. |
Pretreat. with CaCO3 15° CH(3) |
Graph n° 3
|
Pretreat. with CaCO3 30° CH dyn. |
Pretreat. with CaCO3 30° CH (4) |
(1) 10 hours before in phosphate tampon 10 mM at pH 6,5 with the addition of a watery solution of CaCO3 dynamized at 5°CH; after this time the solution of IAA 10-4 M was added.
(2) 10 hours before in phosphate tampon 10 mM at pH 6,5 with the addition of a watery solution of CaCO3 5°CH, previously heated to a temperature of 60° and then left to cool; after this time the solution of IAA 10-4 M was added.
(3) 10 hours before in phosphate tampon 10 mM at pH 6,5 with the addition of a watery solution of CaCO3 15°CH, previously heated to a temperature of 60° and then left to cool; after this time the solution of IAA 10-4 M was added.
(4) 10 hours before in phosphate tampon 10 mM at pH 6,5 with the addition of a watery solution of CaCO3 30°CH, previously heated to a temperature of 60° and then left to cool; after this time the solution of IAA 10-4 M was added.
EXPERIMENTAL PROTOCOL OF POINT 2
Graph. n° 4
|
Pretreat. with CaCO3 5° CH |
Pretreat. with CaCO3 15° CH |
|
Pretreat. with CaCO3 30° CH |
Control |
Graph. n° 5
|
Control |
Treated with IAA 10-4 M |
Discussion
Graph. n° 1
The percentage increase in length was negligible in the control specimens and the dynamized control specimens which contained only solutions of tampon phospate 10 mM at pH 6,5.
Whereas in the specimens simple and dynamized, treated with IAA 10-4 M, a growth of 15-24 % was noted as of the second hour of treatment, reaching 35-45 % by the 9th - 10th hour of treatment.
In the specimen treated with dynamized IAA, an increase of 3-5 % more then in the specimen treated with simple IAA was observed. This is not to be considered statistically significant, however the hypothesis of an improved action whith dynamized IAA is not to be exeluded.
Graph. n° 2
The specimens pretreated 10 hours previously with CaCO3 5°CH, to which was then added IAA 10-4 M, achieved, from the 6th hour onwards, an increase of 8-9% more than the specimen treated with simple IAA. Compared to the specimens treated with dynamized IAA, the above mentioned specimens achieved an increase of 5% from the 6th hour, increasing to 10% more at the 7th hour, and then stabilizing at 4% at the last hour.
The specimens treated 10 hours previously with CaCO3 in 15°CH, to wich IAA 10-4 M was then added, achieved an increase of from the 8th hour to the 10th hour.
This increase would not be statistically significant unless compared with the data of the specimens pre-treated 10 hrs. previously with CaCO3 in 5°CH and 15°CH that had been heated to 60°, and to which IAA 10-4 was added. Infact, the increase noted in the first specimens as compared to that noted in these later specimens was of 9 % and therefore of some, albeit limited, significance.
Graph. n° 3
The specimens pretreated 10 hrs. previously with CaCO3 in 30°CH, to which IAA 10-4 was added, did not achieve a significant increase, ( 3-5% compared to the specimens with IAA), but the increase became significant, from 3-7%, when compared to the specimens pre-treated with CaCO3 in 30°CH but heated to 60°.
Graph. n° 4
The specimens pretreated 10 hrs previouslv with CaCO3 in 5°CH, 15°CH and 30°CH respectively, without the addition of IAA 10-4 M, achieved an insignificant growth compared to those treated exclusivelv with IAA.
Conclusions
From these data it is possible to draw various conclusions, and also new hypothesis for study and research; the conclusions of this research can be summarized as follow:
a) The specimens pretreated 10 hrs previously with a homoeopathic dynamization of CaCO3, which obtained a statistically valid increase when compared with the percentage growth of the specimens with just IAA, were those of 5°CH.
b) The research confirms the hypothesis that the homoeopathic dynamization are neutralized by heating to 60°.7
This can be seen in Graph. n° 2, which shows that the increase obtained in the case of active dynamizations is 9 % more than in the non activated samples.
c) The dynamizations of CaCO3 in 5°CH, 15°CH and 30°CH have not effect on the growth in length of coleoptile of oats isolated from the sub-apex area. Then therefore do not have an activity similar to that of the auxine.
d) The action of dynamizations of CaCO3, above all in 5°CH, is strictly synergic and seems to strengthen the phito-hormonal action of IAA 10-4 M.
Hypothetical considerations
The hypothesis has been formulated that the effect of the auxina on growth is largely the consequence of an acidification of the extra-cellular space. The acidification effect induced by the auxina seems to depend on the stimulation of secret protons through the cellular membrane.
The electrogenic extrusions of protons is linked to a stimulus of the velocity of assumption of cations and in particular of K+.
The correlation between the stimulus of growth in lenght and the activation of the electrogenic system of exchange of H+/K+ suggest that the acidification of the cellular walls plays an important role in the increase of the plastic-extendability induced by the hormones.2
The hypothesis can thus formulated that the role of CaCO3 can be that of:
a) increasing the extracellular Ca++ favouring the activation auxine of the electrogenic system of exchange of H+/K+.5
b) allowing for a better reception of calcium from the culture ground, provoking a consequent increase in the functionality of the ion at cellular level. For Kirby et al. (1984), there exists a precise relatioship between endogenous IAA and the calcium ion.5
This is validated by observation that compounds, such as TIBA, capable of obstructing the auxinic transport, also decrease that of Ca++, causing the symptoms typical of calcium deficiency.8
References
1) Torpe T.A. Organogenesis in vitro: structural, physiological and biochemical aspects. Int. Rev. Cytol. Suppl. II A: 71-111. 1980.
2) Rayle-Cleland. Current Topics in Developmental Biology, 11,1977.
3)Goldsmith M.H.M. The transport of auxin. Ann. Rev. Pl. Physiol. 347-357, 1968.
4) Ileide O.M. Auxin level and regeneration of Begonia leaves. Planta, 81: 153-159, 1968.
5) Kirkby et al. Calcium ions mouvements in the growthing tissues. 1984.
6) Gautheret R.J. Culture des tissues vegetaux: techniques et realisations. Masson et Cie. Paris 1959.
7) Boiron J. Recherche fondamentale en Homeopathie. Experimentation de laboratoire sur l'animal et le vegetal. Encycl. Med. Chir. Paris 1970.
8) Marschner H.; H.M.Haussling; C.A.Jors; G.Lehmbecker; C.H.Hecht-Buchholz. Ion and water uptake in relation to root development in norway spruce (Picea abies). Plant physiol. Vol. 133, pp 486-491. 1988.
Adress for correspondence
Prof. Dr. Bornoroni Corrado
Istituto Superiore di Medicina Olistica e di Ecologia
Università degli Studi di Urbino
Via Saffi, 2
61029 URBINO (PS) Italy
e-mail ismoe@bib.uniurb.it