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 Table of Contents 
ORIGINAL ARTICLE
Year : 2013  |  Volume : 36  |  Issue : 1  |  Page : 38-44  

Effect of gamma irradiation on germination, growth, and biochemical parameters of Terminalia arjuna Roxb


1 Department of Applied Botany, Mangalore University, Mangalagangothri, Karnataka, India
2 Center for Application of Radioisotopes and Radiation Technology, Mangalore University, Mangalagangothri, Karnataka, India
3 Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Center, Trombay, Mumbai, Maharashtra, India

Date of Web Publication21-Nov-2013

Correspondence Address:
K R Chandrashekar
Department of Applied Botany, Mangalore University, Mangalagangothri - 574 199, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0972-0464.121826

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  Abstract 

The impact of gamma irradiation on Terminalia arjuna (T. arjuna), one of the potent medicinal plants for cardiac disease is described in this article. The seeds of T. arjuna were irradiated with different doses of gamma radiation ranging from 0 to 200 Gy using the 60 Co source. The effect of gamma radiation on the growth and biochemical constituents were compared with the control plants. Germination speed at 25 Gy was found to be 0.65, which was double compared to the un-irradiated seeds. An increase in germination percentage, vigor index, and relative growth rate, in terms of dry weight was noticed at lower doses of the radiation treatment. The proline content increased with increasing doses. The chlorophyll content was found to have increased to 12.2 mg/g FW at 100 Gy compared to the control level of 8.44 mg/g FW. Increased phenolic content and radical scavenging capacity was observed at 25 and 150 Gy. Hence, lower doses of radiation treatment may be used to increase the germination, growth, and vigor, and also the enhancement of plant metabolites like proline and phenolics in T. arjuna.

Keywords: Gamma irradiation, germination, growth, proline, Terminalia arjuna


How to cite this article:
Akshatha, Chandrashekar K R, Somashekarappa H M, Souframanien J. Effect of gamma irradiation on germination, growth, and biochemical parameters of Terminalia arjuna Roxb. Radiat Prot Environ 2013;36:38-44

How to cite this URL:
Akshatha, Chandrashekar K R, Somashekarappa H M, Souframanien J. Effect of gamma irradiation on germination, growth, and biochemical parameters of Terminalia arjuna Roxb. Radiat Prot Environ [serial online] 2013 [cited 2020 Jul 7];36:38-44. Available from: http://www.rpe.org.in/text.asp?2013/36/1/38/121826


  Introduction Top


Terminalia arjuna Roxb. (T. arjuna) is an evergreen forest tree, belonging to the family Combretaceae. It produces large orthodox seeds as propagules. The tree is found in the tropical and sub-tropical parts of India and plays an important role in the sericulture industry. [1] Its leaves form an ideal food for the Tasar silkworm Antheraea mylitta. [2] Its white-to-pinkish-gray bark has been used in India's native Ayurvedic medicine for over three centuries, primarily as a cardiac tonic and also as a potent antioxidant for ischemic heart diseases. [3] Wild researches on T. arjuna revealed antioxidant, anti-ischemic, antihypertensive, and antihypertrophic effects, which have relevance to its therapeutic potential in cardiovascular diseases, in humans. Several clinical studies have reported its efficacy, mostly in patients with ischemic heart disease, hypertension, and heart failure, besides its potent antibacterial and antimutagenic activities, with ethnomedicinal importance. [4],[5],[6] Propagation of T. arjuna is difficult by conventional methods, due to poor seed germination and seedling viability. Hence, cuttings and air-layering methods were adopted for this plant. [7] Seed treatments with chemicals such as sulfuric acid and growth hormones like gibberellic acids were used for enhancing germination. [8]

The economical and more effective features of gamma rays due to their high penetration power helps in their wider application for the improvement of various plant species compared to other ionizing radiations. [9] Gamma irradiation has a profound influence on plant growth and development by inducing genetical, cytological, biochemical, physiological and morphogenetic changes in cells and tissues depending on the levels of irradiation. [10] The material and energy necessary for initial growth is already available in the seed, but some stimulants are required only to activate those substances already stored in the cotyledons. Low doses of γ-radiation may increase the enzymatic activation and awakening of the young embryo, which results in stimulating the rate of cell division and affects not only germination, but also vegetative growth. [11]

The biological effect of gamma radiation is mainly due to the formation of free radicals by the hydrolysis of water, which may result in the modulation of an antioxidative system, accumulation of phenolic compounds and chlorophyll pigments. [12],[13],[14],[15] Treatment of crop varieties with gamma radiation has been found to alter the germination and accumulation of proline content resulting in the development of stress-tolerant varieties, along with enhancement in the crop yield. [16] Use of biotechnological approaches with low-dose irradiation treatment for enhancing the production of bioactive plant metabolites, such as, phenolic compounds, salicylic acids, coumaric acids, caffeic acids, flavonoids, and anthocyanins has been documented in medicinal plants. [17]

Activation of the antioxidant system under the varied levels of gamma rays was helpful in understanding the radiosensitivity of a species, which could be used as an ecological indicator. [18],[19],[20] Production of secondary metabolites by the irradiation treatment of a callus was performed by a few investigators for the enhanced production of shikonin-like phenolic acid derivatives. [21] Gamma rays, being oxidative triggers enhanced the production of the medicinally valuable psoralen content along with other biochemical changes, which were helpful in the up regulation of biosynthetic pathways. [22] Enhancement of germination and growth along with plant metabolites, using irradiation technology, could be employed for improving the quality of medicinally valuable plants, including their biomass production.

Despite the usefulness of ionizing radiation to increase the germination potential and generating useful mutations in agricultural crops, there are not many references in the literature on the use of nuclear techniques on forest tree species. [23] Hence, the present study is undertaken to scrutinize the effects of gamma rays on the germination of seeds, growth, and biochemical attributes of seedlings.


  Materials and Methods Top


Mature seeds of T. arjuna were collected from the Western Ghats in the month of May 2011. About 400 clean seeds with moisture content of 5.1685 ± 0.98% were subjected to different doses (25, 50, 100, 150, 200 Gy) of gamma radiation using a Co 60 gamma source (Board of Research in Radiation and Isotope Technology, Mumbai). Dose selection was based on our previous study on the Terminalia (T. chebula unpublished data) species, where a steep decrease in germination and growth attributes were observed above the dose of 200 Gy. Each treatment was replicated five times with 10 seeds in each replicate and arranged in a randomized complete block design. All the seeds were kept on a sand bed for germination for 60 days in the green house (Department of Applied Botany, Mangalore University).



The data were analyzed using IBM SPSS20 statistical software (SPSS Inc. Chicago, IL, USA) following a randomized, complete block design. The mean values were compared using Duncan's Multiple Range Test (DMRT) at 0.05% level of probability.


  Results and Discussion Top


Effect of gamma irradiation on germination and growth characteristics

Results on the effect of gamma rays on germination and other morphological traits are presented in [Table 1]. The seeds irradiated with 25, 50, 100, and 200 Gy have shown significant increase in the percentage of germination compared to the controls, with the highest being observed at 100 Gy. There was no significant difference in the shoot length between the control and the irradiated seedlings. Increase in the number of leaves was clearly evident and the maximum increase was 43% at 200 Gy. Dry weights of the plants were found to be significantly higher at 25 and 50 Gy. It was noticed that the shoot length of the plant remained almost the same but there was a slight increase in the root length and number of leaves in irradiated plants. In Hibiscus sabdariffa L., the enhancement in fresh weight of roots as well as leaves was observed, when treatmed with gamma radiation. [30] Furthermore, a significant increase in dry weight of the whole plant irradiated with 25 and 50 Gy was observed.
Table 1: Effect of different doses of gamma irradiation on the germination and growth parameters of Terminalia arjuna Roxb.

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In fact, many researchers have reported the effect of ionizing radiation on germination, growth, morphology, and yield in different plant species. [31],[32] A three-fold increase in germination was observed by Maherchandani [33] in a dormant Avena fatuva seed exposed to 100 Gy. Improvement in the seed germination at lower doses was observed when genetically pure seeds of Tectona grandis were treated with 100, 200, and 300 Gy doses of gamma radiation. [34] There was an increase in the germination percentage of Hyoscyamus muticus with an increase in gamma irradiation up to 100 Gy, but it decreased thereafter. [35] However, the present study is the first report on the effect of gamma irradiation on T. arjuna, where a bio-positive effect was found with a low dose of radiation treatment. Seeds irradiated with 25, 50, 100, and 200 Gy had shown significant increase in the percentage of germination compared to the control, which was in accordance with the earlier reports on Acacia leucophloea, Albizia lebbeck, and Zizyphus mauritiana. [36] There was no significant difference in the shoot length and root length between the control and irradiated seedlings. In two species of Pinus, an increased dose of radiation above 100 Gy reduced the shoot epicotyls and the root primordia, indicating its sensitivity toward the gamma rays. [37] However, T. arjuna was found to be more resistant to ionizing radiation and significant alteration was not observed in this. Increase in the number of leaves in plants of irradiated seeds was also clearly evident from the table and the maximum increase was about 43% at 200 Gy, which was significantly higher than the control. Enhancement in the dry mass production of Soybean plants was observed when subjected to lower dose of 25 Gy dose. [9] A similar increase in the biomass of plants obtained from the irradiated seeds of T.arjuna with doses of 25 and 50 Gy was also noticed in the present study.

The effect of gamma radiation on the speed of germination is presented in [Figure 1]. It was seen that the speed of germination was two-fold higher than the control and was maximum in the seeds irradiated with 100 Gy followed by the plants of seeds irradiated with 25, 50, 200, and 150 Gy, respectively. The vigor index, a product of the total plant length and percentage of germination is presented in [Figure 2]. The highest vigor index (1522.1) was observed for the plants of seeds irradiated with 100 Gy followed by the plants of seeds irradiated with 25, 50, 150, and 200 Gy respectively. Both, increase in the speed of germination and vigor index are orders of magnitude higher compared to the control. A similar type of increase in the speed of germination and vigor index of gamma irradiated seeds was observed at 100 and 200 Gy in tomato and okra seeds. [38],[39] This was probably due to the fact that short-wave photons (i.e., gamma-rays) were more energetic than visible light photons (>400 nm), and hence, had a stronger effect on the surface of plant cells. This caused the ultimate breakdown of the seed coating allowing the germination to accrue. [12],[40] A significant increase in the germination index, which was the speed of emergence of seedlings, in three Amaranth lines was observed after pre-sowing irradiation treatment. [40] In Pterocarpus santalinus a lower dose of 25 and 50 Gy significantly increased the germination speed and vigor compared to the control. [41] Increase in germination speed would be helpful in increasing the growth rate of the plant, which had a direct influence on the growth of the plant and its successful establishment. The results of present study are also in agreement with these reported literature values.
Figure 1: Effect of different doses of gamma irradiation on the speed of germination in Terminalia arjuna Roxb. Bars showing the same letter are not significantly different at P ≤ 0.05. The data shown are mean ± SD of five replicates. Error bars (i) show SD

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Figure 2: Effect of different doses of gamma irradiation on the vigor index of Terminalia arjuna Roxb. Bars showing the same letter are not significantly different at P ≤ 0.05. The data shown are mean ± SD of five replicates. Error bars (i) show SD

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Effect of gamma irradiation on the biochemical attributes

The estimation of important biochemical qualities such as chlorophyll content, total carbohydrates, and DPPH scavenging activity are presented in [Table 2]. Photosynthesis is one of the most studied processes under the effects of gamma irradiation, accompanied mainly by growth experiments. Significantly higher chlorophyll contents have been observed in seedlings treated with radiation compared to the control. A gradual increase in the chlorophyll content has been observed in the present study, which reached the maximum level at 100 Gy and decreased slightly thereafter. However, no significant increase has been observed in both total carbohydrate and DPPH scavenging activity levels.
Table 2: Effect of different doses of gamma irradiation on total chlorophyll, total carbohydrate, and DPPH radical scavenging activity of Terminalia arjuna Roxb.

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An increase in chlorophyll a, b, and total chlorophyll levels was observed in Paulownia tomentosa plants exposed to gamma irradiation.[42] Plants irradiated at 16 Gy showed a significant increase in their chlorophyll content, which correlated with the stimulated growth in red pepper. [12] Radiation treatment (20 Gy) to dry seeds of lupine increased the total chlorophyll content, soluble sugars, and photosynthetic activity. [43]

Proline being a cytosolic osmoticum and a scavenger of hydroxyl radicals, can stabilize the structure and function of macromolecules such as DNA, protein, and also of the membranes. [44] Proline, sharing this property with other compounds collectively referred to as 'compatible solutes,' are accumulated by a wide range of organisms to adjust cellular osmolarity. [45] Increase in proline content can be helpful in maintaining osmoticum under various environmental stresses. In the present study, the proline content increased with an increase in the dosage of radiation [Figure 3]. The highest concentrations of 0.0418 mg/g FW and 0.04017 mg/FW were recorded in seedlings treated with 100 and 200 Gy respectively. In the present study the proline content of T.arjuna has been found to be peak at 100 Gy and all other doses showed a significantly higher quantity compared to the control. Ionizing radiation treatment of Phoenix dactylifera L with X-rays has significantly increased the proline content of seedlings to overcome radiation-induced stress.[46] Increase in proline content with increased doses of radiation in the present study confirms the role of proline as a compatible solute.
Figure 3: Effect of different doses of gamma irradiation on the proline content of Terminalia arjuna Roxb. Bars showing the same letter are not significantly different at P ≤ 0.05. The data shown are mean ± SD of five replicates. Error bars (i) show SD

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The variations of phenolic concentrations in the seedlings are presented in [Figure 4]. The highest concentration of phenolic content (0.3177 mg/mg) was observed in seedlings irradiated with 150 Gy followed by the seedlings irradiated with 25 Gy (0.31003 mg/mg). Gamma irradiation is known to increase the activity of phenylalanine ammonia-lyase which is responsible for the synthesis of polyphenolic acids. [47] Increase in the production of phenolics was observed in Pterocarpus santalinus when subjected to different doses of gamma radiation. [42] The T. arjuna seedlings exposed to increasing radiation doses up to 150 Gy showed a higher quantity of phenolic compounds, which are a major component of the plant secondary metabolites, with medicinal importance.
Figure 4: Effect of different doses of gamma irradiation on phenolic content of Terminalia arjuna Roxb. Bars showing the same letter are not significantly different at P ≤ 0.05. The data shown are mean ± SD of five replicates. Error bars (i) show SD

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  Conclusion Top


Enhanced germination percentage, speed, and relative growth rate in terms of dry weight and vigor were observed in the seedlings germinated from irradiated seeds. The increased synthesis and accumulation of proline and phenolics suggests the development of a protective mechanism in T. arjuna for better tolerance of plants to radiation-induced stress. Based on the results obtained, it may be concluded that lower doses of radiation may facilitate better germination, growth, and development by overcoming all the barriers including dormancy in T.arjuna.


  Acknowledgments Top


Authors are grateful to BRNS (Board of Research in Nuclear Sciences) for financial support and Mangalore University for providing research facilities. Authors also acknowledge BRIT (Board of Research in Radiation & Isotope Technology Mumbai) for providing Irradiation facility.

 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2]


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