156 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) ISSN (Print) 2313-4410, ISSN (Online) 2313-4402 © Global Society of Scientific Research and Researchers http://asrjetsjournal.org/ Effects of Microwave Radiations on the Morphological and Biochemical Aspects of Some Economically Important Herbs Khan Aisha Saleema*, Butt Sabab, Akhtar Sameera Arzooc a,bDepartment of Biological Sciences, Forman Christian College, Lahore, Pakistan cKinnaird College for Women, Lahore aEmail: aishasaleemkhan@fccollege.edu.pk, bEmail: sababutt@fccollege.edu.pk cEmail: neelofer_arzoo@hotmail.com Abstract The purpose of present work was to observe the effects of microwave radiation on some economically important herbs by treating them with microwaved water. Plants studied were Brassica campestris, Lycopersicon esculentum, Pedilanthus tithymaloides, Portulaca grandiflora, Solanum melongena and Zinnia elegans. The results showed that microwave treated water and warm water both inhibited the growth of treated plants as there was significant decrease in the root and shoot length, diameter, fresh and dry weight, reduction in chlorophyll content, leaf area and enzyme activity of peroxidases which was also altered in all the treated plants. It was reported that the microwave radiations were harmful to treated plants and therefore the use of microwave appliances for heating purposes can affect the biochemical nature of food and ultimately can have an affect on human health. Keywords: Brinjal; Microwave; Morphology; Mustard; Peroxidase; Purslanes. 1. Introduction Harmful effects of radiations have been reported since long on plants [1,2,3,4,5]. They are known to cause chromosomal aberrations, damage to DNA, inhibition in seed germination and can also damage exposed tissues [6, 2]. Microwaves radiations also have an effect on the enzymatic activities. They are know to affect the enzyme function, cause cell dysfunction and death [7,8]. Peroxidases are involved in many important metabolic processes in plants and are considered to be more regulated under stress conditions, contribute to the defense of plants against pathogens [9]. and to wound healing [10]. They are localized mainly in the cell wall and in vacuoles. Plant roots exude high concentration of peroxidases in soil, particularly in response to water and chemical stress [11]. ------------------------------------------------------------------------ * Corresponding author. http://asrjetsjournal.org/ American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 157 Plants peroxidases have been implicated in a number of diverse phenomena observed in plants i.e., lignification, suberization, cell elongation, growth and regulation of cell wall biosynthesis and plasticity. They have been shown to be useful as markers of the reaction of plants to external environmental stresses. 2. Materials and Methods 2.1. Experimental setup /area of work Three sets of each plant were selected for this work. They were grown in pots in the botanical garden of Kinnaird College, Lahore. One set was treated with tap water and was taken as control, second set was treated with microwave water (1 minute time), and then placed at room temperature for few minutes. It was then applied to one set of all plants. Third set of plants was treated with tap water but heated for few minutes and then cooled at room temperature before applying. 2.2. Water Treatments Each set was given 300 ml of water of each type. All plants were grown outdoors under natural day light in equal sized earthen pots. Plants selected were of almost same age. They were treated with microwave and warmed water for one month. 2.3. External and Internal Morphology For external and internal morphology, length and diameter of root and shoot were calculated. Minitab V (13) Mean, standard error of the mean, one way ANOVA were calculated. 2.4. Estimation of Enzymatic Activities (Peroxidases) In a cold pastle and mortar, weighed and frozen plant material was crushed with phosphate buffer (0.1M; pH 7) (Table 4.1.12) in the ratio of 1:4 (w/v) i.e., 1 gm of plant material; 4 ml of phosphate buffer. The samples were centrifuged at 10,000 rpm for 10 minutes. The supernatant was used for estimation of peroxidases. Two sets of test tubes were labeled (one for experimental and one for control). In all the test tubes (2 sets) 2.5 ml of phosphate buffer (pH 7.0) 0.2 ml of enzyme extract was added. In experimental set 0.2 ml of 1% Guaiacol solution was added and mixed. Both the sets were left at room temperature for 15-20 minutes. Then 0.1 ml of 0.3% H2O2 was added in all the test tubes and stirred. For blank, 0.2 ml of glass distilled water, 2.5 ml of phosphate buffer (pH 7.0) and 0.1 ml of 0.3% H2O2 was mixed. Optical density of all the test tubes was taken against this test tube for blank. The absorbance was taken at 750 nm on a Beckmann 200D spectrophotometer. Formula used for peroxidases were as follows: Units mg-1 = O.D of experimental – O.D. of control O.D of control x mg of fresh plant material 3. Results 3.1. External Morphology American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 158 In the present study, the microwaved water treated plants showed a marked decrease in the growth of the roots, shoots and leaves. The length and diameter of the root and shoot of the microwaved treated plants was highly affected (Table 1-7, fig. 1-6). The number of flower buds was also affected as plants treated with the microwaved water had lesser number of flower buds as compared to the control plants (Table 1-7). Table 1: Effects of Microwave Radiations on root parameters of treated plants (Readings are the mean of five replicates) Treated Plants length of root (cm) Treatments Control Warm water Microwaved water Brassica campestris 4.25±0.033 4.45±0.666 3.89±0.667 Lycopersicon esculentum 6.24±0.122 3.98±0.089 2.88±0.080 Pedylanthus tithymaloides 7.21±0.334 6.55±0.045 4.86±0.031 Portulaca grandiflora 6.88±0.566 4.98±0.001 3.4±0.044 Zinnia Elegans 4.5±0.045 5.09±0.0021 2.29±0.012 Solanum melongena 5.12±0.069 4.64±0.023 4.60±0.012 Table 2: Effects of Microwave Radiations on shoot of Brassica campestris (Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot (cm) No. of buds Appearance of flowering Control 13.3±0.045 0.04±0.067 3 15th day Warm water 9.05±0.091 1.046±0.012 2 10th day Microwaved water 9.06±0.002 1.043±0.089 2 12th day Table 3: Effects of Microwave Radiations on shoot of Lycopersicon esculentum (Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot (cm) No. of buds Appearance of flowering Control 11.4±0.98 1.038±0.67 0 0 Warm water 11.4±0.67 1.033±0.12 0 0 Microwaved water 6.3±0.055 1.034±0.04 0 0 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 159 Table 4: Effects of Microwave Radiations on shoot of Pedylanthus tithymaloides (Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot (cm) No. of buds Appearance of flowering Control 38.4±0.455 1.0885±0.554 0 0 Warm water 35.5±0.002 1.081±0.988 0 0 Microwaved water 33.5±0.129 0.065±0.001 0 0 Table 5: Effects of Microwave Radiations on shoot of Portulaca grandiflora (Readings are the mean of five replicates)(Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot No. of buds Appearance of flowering Control 19.05±0.778 1.83±0.788 9 10th day Warm water 20.3±0.233 1.04±0.234 15 15th day Microwaved water 15.6±0.023 1.032±0.124 17 20th day Table 6: Effects of Microwave Radiations on shoot of Solanum melongena (Readings are the mean of five replicates) (Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot (cm) No. of buds Appearance of flowering Control 17.7±0.078 2.95±0.680 0 0 Warm water 13.2±0.124 2.03±0.347 0 0 Microwaved water 11.4±0.045 1.83±0.246 0 0 Table 7: Effects of Microwave Radiations on shoot of Zinnia elegans (Readings are the mean of five replicates) Treatments Length of shoot (cm) Diameter of shoot (cm) No. of buds Appearance of flowering Control 25.5±0.687 2.645±0.689 6 14th day Warm water 20.3±0.123 2.034±0.002 2 16th day Microwaved water 21.5±0.045 2.032±0.001 2 21st day American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 160 Figure 1: Effects of different treatments on root length of B. Campestris Figure 2: Effects of different treatments on root of L. esculentum Figure 3: roots of different treatments of Zinnia elegans in comparison American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 161 Figure 4: Effects of different treatments on l. Esculentum Figure 5: Effects of different treatments on p. Grandiflora Figure 6: Effects of different treatments on z. Elegans American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 162 3.2. Internal Morphology In all the treated plants, significant increase in peroxidase activities was observed in all microwaved treated plants as compared with control. Peroxidase activity was more pronounced in roots of all plants (Table 8-13). Table 8: Effects of microwaves on the peroxidase activity in b. Campestris Part (mg-1) Treatments Control Warm water Microwaved water Root (mg-1) 2.58±0.003 2.05±332.3 3.03±564.31 1st internode (mg-1) 2.18±0.033 2.06±497.9 2.98±25.76 1st leaf (mg-1) 2.09±0.009 1.98±68.07 12.45±89.34 s Table 9: Effects of microwaves on the peroxidase activity in l. Esculentum Part Treatment (ml) Control Warm water Microwaved water Root (mg-1) 4.67±26.09 4.12±37.21 5.67±47.07 1st internode (mg-1) 3.12±55 3.34±106.54 3.87±188.78 1st leaf (mg-1) 1.16±0.95 1.34±1.80 1.87±42.15 Table 10: Effects of microwaves on the peroxidase activity in p. Tithymaloides Part Treatment (ml) Control Warm water Microwaved water Root (mg-1) 1.67±34.15 1.12±31.98 2.67±27.19 1st internode (mg-1) 1.12±23.09 1.34±12.98 1.87±89.12 1st leaf (mg-1) 0.16±4.12 0.44±2.19 0.93±56.08 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 163 Table 11: Effects of microwaves on the peroxidase activity in p. Grandiflora Part Treatment (ml) Control Warm water Microwaved water Root (mg-1) 5.12± 12.08 4.92±12.89 6.12±49.12 1st internode (mg-1) 2.12±6.78 2.46±36.512 2.93±40.78 1st leaf (mg-1) 0.18±8.17 0.81±34.80 0.93±83.12 Table 12: Effects of microwaves on the peroxidase activity in s. Melongena Part Treatment (ml) Control Warm water Microwaved water Root (mg-1) 3.75± 12.70 4.32±10.67 4.92± 12.89 1st internode (mg-1) 3.59±4.78 4.46±8.35 4.93±12.98 1st leaf (mg-1) 2.67±9.34 2.81±67.43 1.85±15.78 Table 13: Effects of microwaves on the peroxidase activity in z. Elegans Part Treatment (ml) Control Warm water Microwaved water Root (mg-1) 5.12± 2.89 6.81±45.11 7.12± 33.71 1st internode (mg-1) 4.12±9.23 4.10±5.89 5.17±63.12 1st leaf (mg-1) 1.75±5.12 1.89±61.2 2.85±10.54 4. Discussion The treatment of plants with microwaved water resulted in significant inhibition of plant growth as compared with untreated control plants (Table; 1-7, Fig. 1-6). In all the treated plants, many inter and intraspecific differences were observed in response to microwave radiations with respect to growth of root, shoot and leaves. Similar results are reported by many workers [12, 8] with plants under stress. Among the treated plants L. American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 164 esculentum, B. campestris and Z. elegans showed remarkable differences as compared with P. grandiflora. Further, flowering in all the microwaved water treated plants was also delayed although bud initiations was observed but yet the flowering was delayed (Table 1-7). Plants under stress caused delay in flowering as flower formation is related to environmental conditions [13,14]. Flower development involves a complex interaction of molecular, biochemical, and structural changes. However, little information is available on the physiology of early flower development, on the molecular aspects of fruit development in general, and on how flower development is coordinated with hormonal action. Plants under stress caused delay in flowering as flower formation as it is related to different environmental conditions [13]. Stress caused inhibition in flowering can be attributed to initiation of disruption in biological processes [8]. In all the treated plants, increase in enzyme activity was observed (Table 8-13), further indicating the role of radiations in inducing stress in different parts of plants. The role of peroxidase as a stress enzyme in plants has been widely accepted [15]. Maximum enzyme activity was detected in roots, then in shoot and in leaves. This might be due to different level of stress of tolerance different plants [16]. Roots of plants were more responsive to stress because peroxidase activity was maximum in the roots of treated plants, showing the sensitivity of roots to microwaved radiations. 5. Conclusions The harmful effects of microwave radiation was reported in all the treated plants.The results were correlated to the facts that radiations can also cause damage to human tissues. Different plants were treated in order to observe the effects of microwave radiations. Changes caused by radiation were reported to be inhibitory for all the treated plants showing that harmful radiations can also interact with the enzymes causing a disruption of biological processes in plants. 6. Recommendations This research work reports that effects of microwaved radiations cause inhibitory effects on the growth of all treated plants so it is suggested that microwaved radiations are not good for human health and use of microwave in everyday life should be minimized as microwaved radiations change the nature of food and cause abrupt biological changes. References [1]. A. Vian, E. Davies, M.Gendraud, and P. Bonnet (2016).' Plant Responses to High Frequency Electromagnetic Fields'.BioMed Research International. http://dx.doi.org/10.1155/2016/1830262. [2]. JL. Ryan (2012). "Ionizing Radiation: The Good, the Bad, and the Ugly," J. Invest. Dermatol.132, p.985. [3]. Z. Forgacs, G. Kubinyi, and G. Sinay (2005). ''Effects of 1800 MHz GSM-like exposure on the gonada function and hematological parameters of male mice''. Magy Onkol. 49(2), p. 149–151. [4]. D. Sripakdee, KL. Sukontason, S. Piangjai, R. Ngern-klun, and K. Sukontason (2005). ''Effect of http://dx.doi.org/10.1155/2016/1830262 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2019) Volume 51, No 1, pp 156-165 165 microwave irradiation on the blow fly Chrysomya megacephala (F.) (Diptera: Calliphoridae)''. Southeast Asian Journal of Tropical Medicine and Public Heslth. 36(4), p. 893–895. [5]. LW. Barnthouse. (1995). "Effects of Ionizing Radiation on Terrestrial Plants and Animals: a Workshop Report," Oak Ridge National Laboratory, ORNL/TM-13141. [6].BW, Shirley, S. Hanley, and HM. Goodman (1992). "Effects of Ionizing Radiation on a Plant Genome: Analysis of Two Arabidopsis transparent testa Mutations," Plant Cell. 4, p. 333. [7].T. Ozaki, K. Tabuse, T. Tsuji, Y. Nakamura, K Kakudo, and I. Mori (2003). ''Microwave cell death: enzyme histo-chemical evaluation for metastatic carcinoma of the liver.'' Pathology International. 53(12), p/ 837–845. doi: 10.1046/j.1440-1827.2003.01571.x. [8]. P.Sharma , and RS. Dubey. (2005).'' Lead toxicity in plants''. Brazilian Journal of Plant Physiology.17, p.105-121. [9]. M. Hrubcova, M. Cvikrov, L. Meravy, J. Eder, and P. Binarova. (1992). ''Phenolic accumulation and peroxidase activity in in vitro selected alfalfa callus cultures resistant to filtrate of Fusarium spp''. Biologia Plantarum. 34, p. 203-211. [10].CG. Smith, MW. Rodgers, A Zimmerlin, D Ferdinando, and GP Bolwell. (1994). ''Tissue and subcellular immunolocalisation of enzymes of lignin synthesis in differentiating and wounded hypocotyl tissue of French bean (Phaseolus vulgaris L.)''. Planta.192, p. 155-164. [11]. T. Hirata, Y. Ashida, H. Mori, D. Yoshinaga, and L.J. Goad. (2000). ''A 37kDa Peroxidase secreted from liverworts in response to chemical stress''. Phytochemistry. 55, p. 197-202. [12]. I.R. Mor, S.J. Gokani, and S.V. Chanda. (2002). ''Effect of mercury toxicity on hypocotyls elongation and cell wall loosening in Phaseolus seedlings''. Journal of Plant Nutrition. p, 843-86. [13]. F. Tooke, M. Ordidge T. Chiurugw, and N. Battey. (2005). ''Mechanisms and function of flower and inflorescence reversion'' . Journal of Experimental Botany. 56, p. 2587-2599. [14]. N.Y. Chaudhry, and A.S. Khan. (2006). ''Improvement of pistillate flowers yield with GA3 in heavy metals treated plants''. Plant Growth Regulation. 50, p. 211-217. [15]. K. Subhashini, and GM Reddy. (1990). ''Effect of salt stress on enzyme activities in callus cultures of tolerant and susceptible rice cultivars''. Indian Journal of Experimental Biology. 28, p. 277-279. [16]. A.B. Morek, and M. Wierzbicka (2004). ''Localization of lead in root tip of Dianthus carthusianorum''. Acta Biologica Cracoviensia. 46, p. 45-56. http://large.stanford.edu/courses/2015/ph241/miller1/docs/tm13141.pdf [1]. A. Vian, E. Davies, M.Gendraud, and P. Bonnet (2016).' Plant Responses to High Frequency Electromagnetic Fields'.BioMed Research International. http://dx.doi.org/10.1155/2016/1830262. [2]. JL. Ryan (2012). "Ionizing Radiation: The Good, the Bad, and the Ugly," J. Invest. Dermatol.132, p.985.