|Year : 2014 | Volume
| Issue : 1 | Page : 19-23
Dose-dependent effect of homoeopathic drug Zinc sulphate on plant growth using Bacopa monnieri as model system
Vivek Kumar Gupta1, Jutika Rani Ray1, Vishal Kumar Singh1, Surya Deo Pathak2, Chaturbhuja Nayak3, Mahendra P Darokar1
1 Molecular Bioprospection Department, Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, Uttar Pradesh, India
2 Homoeopathic Drug Research Institute, Viraj Khand, Lucknow, Uttar Pradesh, India
3 Former Director General, Central Council for Research in Homoeopathy, Ministry of Health and Family Welfare, New Delhi, India
|Date of Submission||20-Jun-2013|
|Date of Acceptance||19-Feb-2014|
|Date of Web Publication||29-Mar-2014|
Mahendra P Darokar
Department of Molecular Bioprospection, Central Institute of Medicinal and Aromatic Plants, Kukrailpicnic Spot Road, Lucknow - 226 015, Uttar Pradesh
Source of Support: Central Council for Research in Homoeopathy, Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi,, Conflict of Interest: None
Clinical trial registration not applicable
Background: Zinc is one of the essential micronutrients in plants required in very low quantity for plant growth and development. In higher concentration, it is known to to reduce the rate of photosynthesis, So homoeopathic preparations tested to see it role on plan growth.
Objective: To analyse the effect of homoeopathic preparation of Zinc sulphate on plants through in-vitro assay using Bacopa monnieri as a model plant system.
Materials and Methods: Six homoeopathic potencies (1X to 6X) of Zinc sulphate were used on a decimal scale along with the control (MS basal agar medium). The samples were evaluated by adding fixed amount (100 μl) in the media as well as by dipping the explants in the test sample overnight. At the completion of the incubation period (14 days) the fresh and dry weight, number and length of the roots, number and length of the shoots and the number of leaves were analysed.
Results: It was observed that Zinc sulphate showed growth inhibition at potencies from 1X to 5X, whereas at potency 6X, it exhibited growth promotion effect, when compared with the control.
Conclusion: Homoeopathic drug (Zinc sulphate) exhibited growth promotion at higher potency (6X) and growth inhibition at lower potencies (1X to 5X) on Bacopa monneiri.
Keywords: Bacopa monnieri, Homoeopathic potencies, In vitro assay, Photosynthesis, Ultra molecular, Zinc sulphate
|How to cite this article:|
Gupta VK, Ray JR, Singh VK, Pathak SD, Nayak C, Darokar MP. Dose-dependent effect of homoeopathic drug Zinc sulphate on plant growth using Bacopa monnieri as model system. Indian J Res Homoeopathy 2014;8:19-23
|How to cite this URL:|
Gupta VK, Ray JR, Singh VK, Pathak SD, Nayak C, Darokar MP. Dose-dependent effect of homoeopathic drug Zinc sulphate on plant growth using Bacopa monnieri as model system. Indian J Res Homoeopathy [serial online] 2014 [cited 2019 Nov 16];8:19-23. Available from: http://www.ijrh.org/text.asp?2014/8/1/19/129673
| Introduction|| |
Micronutrients are essential components for balanced growth and development in plants.  Zinc (Zn) is one of the most important essential micronutrients, which is mainly supplied as Zinc sulphate (ZnSO 4 ). It regulates many physiological and a variety of metabolic processes in plants.  Zn plays a vital role in the cell division, cell expansion, protein synthesis and also in carbohydrate, nucleic acid and lipid metabolism.  Besides this, it also plays an important role in auxin metabolism. ,,, This micronutrient is also an essential component of zinc finger transcription factors involved in plant morphogenesis and organogenesis. , However, at much higher concentrations, it can become toxic, thus leading to physiological and morphological disturbances, and eventually decreasing yield. An optimum (3-30 μM) concentration of Zn is required by the plant for growth and development and excess of Zn reduces photosynthetic rate hindering the plant growth. 
The use of homoeopathic preparations in agriculture has started recently. Homoeopathic preparations are being used efficiently for increasing active principles in medicinal plants, plant detoxification for metals such as aluminum and copper, increasing plant growth rate and productivity,  plant metabolism , and control of diseases. ,, Based on these reports, the present study was undertaken to explore the applicability of the homoeopathic preparations Zinc sulphate in some variables of the plant using Bacopa monnieri  as model system for growth and productivity.
| Material and Methods|| |
Homoeopathic preparations of Zinc sulphate in decimal scale (1X to 6X) as per the principles of homoeopathy, was provided by Central Council for Research in Homoeopathy (CCRH), Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi. The study was carried out at CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow during 2007-2010.
A fast propagating strain of Bacopa monnieri (BM08), which showed rapid in vitro response against different compounds/extracts/drugs as well as to physical stresses was used for evaluating effects of homoeopathic drug Zinc sulphate on plants. The testing was carried out by following the standard tissue culture techniques as described earlier. 
The powdered globules Zinc sulphate samples admixed with lactose of different potencies were dissolved in sterile double distilled water. Evaluation of biological activity was performed by two methods in three replicates for each treatment.
In method A, 100 μl of Zinc sulphate sample was mixed with 900 μl of MS basal agar medium and poured into 1.5 ml of graduated micro-centrifuge tube. The twigs (2.5 cm) of in vitro grown Bacopa plants were inoculated in three replicates for each treatment.
In method B, the twigs were dipped in drug samples in three replicates for each treatment overnight and inoculated in 1 ml MS basal agar medium poured into graduated micro-centrifuge tube without drug sample.
These inoculated tubes were kept in half transparent desiccators, allowing the air passage through sterile cotton plugs fixed in the opening vents. The tubes were placed so that the medium containing portion of tubes where roots would be initiated was inserted into the stands, which were made of thermocoal sheet. This set up helps to prevent light exposure to the root while the aerial part of plant got sufficient light and aeration. These desiccators were incubated at normal ambient temperature of 25-28°C with a cycle of 14 hours of light and 10 hours of darkness. Data was recorded in terms of root initiation (number and length), shoot elongation (number and length) and increase in number of leaves for every 24 hours from second day of inoculation till 14 days. MS basal agar medium without any drug sample served as control. Controls provided by CCRH for each potency was also included in the evaluation.
Initiation of roots
Root initiation was observed every 24 hours till the 14 th day. The observations were recorded as roots number and root length in centimeter (cm).
Shoot length was measured by measuring the distance between base of the stem and shoot apex.
Number of leaves
Increase in the number of leaves was recorded at every 24 hours till the 14 th day.
After completion of incubation period (14 days), the fresh weight of plants was determined by weighing on analytical balance. The initial weight of explants was subtracted from final weight.
After completion of incubation period (14 days), the plants were kept in incubator at 37°C for dry weight determination by weighing on analytical balance.
The data for all variables was analysed for mean values and standard error. Statistical analysis was performed by using one way Analysis Of Variance (ANOVA) and was considered at P < 0.05 and P < 0.01 level of significance.
| Results|| |
The treatment of Zinc sulphate in different potencies (1× to 6×) to the plant using Bacopa monnieri as a model system showed different results in in vitro assay [Figure 1]. When zinc sulphate was applied at 1× to 5× potencies, there was a decrease in fresh and dry weight, number and length of roots, length and number of shoots and number of leaves in both the methods of treatment, that is the drug was mixed in the medium and when explant was dipped in the drug. But at 6× potency, root number, root length, number of leaves, fresh and dry weight of plant increased significantly (P < 0.01) in both the methods [Table 1] and [Table 2].
|Figure 1: Dose-dependent effect of Zinc sulphate on plant growth using Bacopa monnieri (a) 100 μl of drug mixed in the MS medium, (b) Explant dipped in the drug|
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|Table 1: Effect of Zinc sulphate in different potencies on Bacopa monnieri using method A|
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|Table 2: Effect of Zinc sulphate in different potencies Bacopa monnieri using method B|
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| Discussion|| |
Since there is decrease in number as well as length of root and shoot, number of leaves, fresh and dry weight of Bacopa plants at 1× to 5× potencies, it is assumed that these potencies of Zinc sulphate are more than the desired that is required for plant growth and development because Zinc sulphate is known to cause phytotoxicity beyond certain concentration. , However, at potency 6× there is an increase in number and length of root and shoot, number of leaves, fresh and dry weight of plants. This might be due to the concentration of Zinc at 6× potency is equivalent to that of the desired concentration of the element for normal plant growth and development. However, potencies more than 6× are needed to be evaluated for conclusive interpretation. Sulphur is an essential macronutrient and required in large amount.  Considering the very low concentration of Sulphur in 6× potency, it would not have any effect on plant growth and development. Therefore, only Zinc was considered since it is an essential micronutrient in plants required in very low amount (3-30 μM) for plant growth and development, which plays a fundamental role in many physiological and biochemical processes, such as structural and functional integrity of bio-membranes, photosynthesis, auxin biosynthesis and metabolism, response to oxidative stress, apoptosis inhibition and phosphate transport. ,,, Zinc is also an essential component of enzymes and transcription factors involved in plant morphogenesis and organogenesis. , Nevertheless, these observations can be substantiated by the fact that homoeopathic drugs also contain the starting material even at higher potencies  and they increase plant metabolism by increasing production and allocation of carbon skeletons that would be used for maintenance and growth.  It was reported previously by Godoy  and Bonato et al.,  that, in Homoeopathy, the same drug frequently causes different effects, depending on the applied dynamisation. Variable effects of different dynamisations of homoeopathic drug on plant Cerotoma tingomarianus have been reported.  Such behaviour of homoeopathic drugs is still insufficiently explained. But it is believed to be related to existing rhythmic movements in nature, and also to the law of similitude that occurs between the applied homoeopathic solution and the organism that receives it. , In homoeopathic science, it is also believed that when there is a similitude between the homoeopathic solution and the organism, the vital energy responds, opposing the energy of the medication to a text of its own energy.  However, when there is no similitude between the homoeopathic solution and the organism, there can be disorder in the metabolic system of the plant, resulting in several negative growth and developmental aspects of the plant. Therefore, the observations made in the present investigation are of great importance with special reference to the use of homoeopathic formulations in agriculture for higher productivity.
| Acknowledgements|| |
The authors are thankful to Central Council for Research in Homoeopathy, Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi for financial support. The authors are also thankful to National Gene Bank for Medicinal and Aromatic Plants, CIMAP, Lucknow for providing plants of Bacopa monnieri.
| References|| |
|1.||Graham RD, Welch RM, Bouis HE. Addressing micronutrient malnutrition through enhancing the nutritional quality of staple foods: Principles, perspectives and knowledge gaps. Adv Agron 2001;70:77-142. |
|2.||Ramesh SA, Choimes S, Schachtman DP. Over-expression of an Arabidopsis zinc transporter in Hordeum vulgare increases short-term zinc uptake after zinc deprivation and seed zinc content. Plant Mol Biol 2004;54:373-85. |
|3.||Collins JC. Effect of Heavy Metal Pollution on Plants. Vol. 1. London and New Jersey: Applied Science Publishers; 1981.p. 145. |
|4.||Hossain B, Hirata N, Nagatomo Y, Akashi R, Takaki H. Internal zinc accumulation is correlated with increased growth in rice suspension culture. J Plant Growth Regul 1997;16:239-43. |
|5.||Saeki Y, Yasukouchi A, Nagatomo Y, Takaki H. Distinctive expression of a zinc binding protein in rice cell callus growth with high zinc concentration. Soil Sci Plant Nutr 2000;46:209-16. |
|6.||Oguchi K, Tanaka N, Komatsu S, Akao S. Characterization of NADPH-dependent oxireductase induced by auxin in rice. Physiol Plant 2004;121:124-31. |
|7.||Oguchi K, Tanaka N, Komatsu S, Akao S. Methylmalonate- semialdehide dehydrogenase is induced in auxin-stimulated and zinc-stimulated root formation in rice. Plant Cell Rep 2004;22:848-58. |
|8.||Kobayashi A, Sakamoto A, Kubo K, Rybka Z, Kanno Y, Takatsuji H. Seven zinc-finger transcription factors are expressed sequentially during the development of anthers in petunia. Plant J 1998;13:571-6. |
|9.||Yanagisawa S. Dof domain proteins: Plant specific transcription factors associated with diverse phenomena unique to plants. Plant Cell Physiol 2004;45:386-91. |
|10.||Ali G, Srivastava PS, Iqbal M. Influence of cadmium and Zn on growth and photosynthesis of Bacopa monniera cultivated in vitro. Biol Plantarum 2000;43:599-601. |
|11.||Bonato CM, Da Silva EP. Effect of the homoeopathic solution Sulphur on the growth and productivity of radish. Acta Scientiarum. Agro Maringá 2003;25:259-63. |
|12.||Betti L, Brizzi M, Nani D, Peruzzi M. Effect of high dilutions of Arsenicum album on wheat seedlings from seeds poisoned with the same substance. Br J Homeopath 1997;86:86-9. |
|13.||Silva MR, Casali VW, Bonato CM, Santos NT. Interaction among CO 2 assimilation and minutes post-treatment of Sphagneticola trilobata with Apis mellifica 6CH. Cultura Homeopática Archivos da Escola de Homeopatia 2006;16:48. |
|14.||Verma HN, Verma GS, Verma VK, Krishna R, Srivastava KM. Homoeopathic and pharmacopeia solutions as inhibitors of tabacco mosaic virus. Indian Phytopath. 1969; 22:188-93. |
|15.||Kumar RS. Effect for certain homoeopathic medicines on fungal growth and conidial germination. Indian Phytopath 1980;33:620-2. |
|16.||Khanna KK, Chandra S. Control of fruit rot caused by Fusarium roseum with homoeopathic solutions. Indian Phytopath 1983;36:356-7. |
|17.||Khanuja SP, Darokar MP, Mishra S, Gangwar A, Shasany AK, Kumar TR, et al. Development of novel plant system as a biosensor for detecting environmental biohazards and bioactive molecule through distinct responses. J Environ Pathol Toxicol Oncol 2001;20:15-22. |
|18.||Berry L, Wallace W. Zinc phytotoxicity: Physiological responces and diagnostic criteria for tissues and solutions. Soil Sci 1989;147:390-7. |
|19.||Montilla M, Parra A, Torrent J. Zinc phytotoxicity to oil seed rape grown on zinc loaded substrate consisting of Iron oxide coated and calcite sand. Plant Soil 2003;257:227-36. |
|20.||Khan TA, Mazid M. Nutritional significance of sulphur in pulse cropping system. Biol Med 2011;3:114-33. |
|21.||Marschner H. Mineral nutrition of higher plants. 2 nd ed. New York: Academic Press; 1995. p. 33-347. |
|22.||Huang C, Barker SJ, Langridge P, Smith FW, Graham RD. Zinc deficiency up-regulates expression of high affinity phosphate transporter genes in both phosphate sufficient and deficient Barley roots. Plant Physiol 2000;124:415-22. |
|23.||Chimienti F, Seve M, Richard S, Mathieu J, Favier A. Role of cellular zinc in programmed cell death: Temporal relationship between zinc depletion, activation of caspases, and cleavage of Sp family transcription factors. Biochem Pharm 2001;62:51-62. |
|24.||Hoseini R, Mulligan BJ. Application of rice (Oryza sativa L.) suspension culture on studying senescence in-vitro (I). Single strand preferring nuclease activity. Electron J Biotechnol 2002;5:42-54. |
|25.||Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homoeopathic dilutions retain starting materials: A nanoparticulate perspective. Homeopath 2010;99:231-42. |
|26.||Bonato CM, de Proenca GT, Reis B. Homoeopathic drug Arsenicum and sulphur affect the growth and essential oil content in mint (Mentha arvensis). Maringa 2005;31:101-5. |
|27.||Godoy M. As potências em homeopatia: Escala de dinamizações de freqüencial ascendente. Revista de homeopatia 1988;53:101-5. |
|28.||Bonato CM, de Proenca GT, Reis B. Homoeopathic drug Arsenicum and sulphur affect the growth and essential oil content in mint (Mentha arvensis) Maringa 2005;31:101-5. |
|29.||Hamly EC. A arte de curar pela homeopatia: o Organon de Samuel Hahnemann. 1 st ed. São Paulo: Prol; 1979. |
|30.||Vithoulkas G. Homeopatia: ciência e cura. São Paulo: Cultrix; 1980. |
[Table 1], [Table 2]