Effects of Microwave Doses on Seed Exposure

personal outlets of Micro cockle Doses on Seed Expo legitimateAbstractA massive increase in electromagnetic pollution since the introduction of telecommunication instruments especially zap oven from which the mobile communication. Current research study take on to assess the physiological effects of inseminate photograph to variant doses of microwave. zap-induced electrolyte fountain, sprouting, chlorophyll and growth were monitored and evaluated following beginning characterisation to microwave from a magnetron of 2.45 GHz, uttermost output personnel of 800 W and wavelength of 12 cm operated at 220 VAC. seeds of genus Hordeum vulgare were exposed to eight different photo periods of microwave from 0 to 600 seconds, experiments were performed in vitro. Percentage of germinated seeds, copulation sprouting coefficient, germination rate, germination index, clear and dry weights, shoot stalk ratio were assessed. Germination parameters were dose-dependents, the parting of germinated seeds were increased later on short image periods to microwave recording 100 % germination. Further the germination rate, sexual relation germination coefficient were as well as increased after short exposure periods to microwave. Longer exposure periods reduced the contri stillion of germination, germination rates and several(a) germination indices. Morphological and growth traits showed a similar trend and were importantly hardend after longer exposure periods to microwave. Chlorophyll contents were world-shakingly decreased with interpolate magnitude exposure periods of microwave. Microwave-induced electrolyte wetting (%) was epochally increased (r=0.92*, pKey words Microwave, electromagnetic, barley, tissue layer ion leakage, cell death, germination, chlorophyll, SPAD, growth, Hordeum vulgare L. basisThe Development of life was regularised by two ubiquitous forces the gravity and electromagnetism, the two forces expected to engender immanent role in th e functional activities of biologic systems and organisms (Balmori, 2009). Previously, microwave radiofrequencies included a few radio and televisions vector located in remote area or high places. A massive increase in electromagnetic pollution since the introduction of telecommunication instruments in the 1990s (Galeev, 2000 Firstenberg, 2001 Ragha et al., 2011) (Ragha et al., 2011). These electromagnetic fields can have a deleterious and damaging effects depending on the exposure doses, power level, frequencies, pulsed or continuous wave and the dielectric properties of exposed tissue, the interaction of such electromagnetic fields on various life processes has been rivet on different microwave frequency range forms an important part (Banik et al., 2003). Microwave are a part of electromagnetic putzs spectrum comprising frequencies ranging from 300 MHz to 300 GHz, further, it act through absorption on molecular(a) level manifesting as vibrational energy or heat and a biologic al effects (Chipley, 1980 Dardanonl et al., 1985, 1994 Pakhomov et al., 1998)(Chipley, 1980 Dardanoni et al., 1985 1994 Pakhomov et al., 2001) including various genetic transmits. Relevant research suggests that microwaves may have long-term health effects (Lin, 2004). Identification, evaluation and judging of the bio-effects of microwaves have been complex and contr everyplacesial, because of the absence of a clear mechanism of the impact and interaction of microwave radiofrequencies and biological systems, there has been a persistent view in biophysical and engineering sciences, that microwave fields are incapable of inducing bio-effects other than by heating (Banik et al., 2003). In recent durations, non-thermal bio-effects of microwaves on tissue responses were being adjudge (Dardalhon et al., 1979a,b Adey, 1981 Banik et al., 2003). heterogeneous research data have offered convincing evidence of non-thermal microwave effects and have similarly indicated various consistenci es in these effects dependency of frequency within specific frequency windows of resonance-type dependency on modulation and polarization dependency on intensity within specific intensity windows, including super-low power parsimony comparable with intensities from base stations/masts (Adey, 1981 Belyaev, 2005 Hyland, 2000 Lai, 2005, (De Salles, 1999 Scialabba and Tamburello, 2002)). Some studies have demonstrated different microwave effects depending on wavelength in the range of mm, cm or m (Kemerov et al., 1999 Nikolaevich et al., 2001). Duration of jibe may be as important as power density (Abu-Elsaoud, 2015), the effect of electromagnetic radiotherapys could be depending on the radiation exposure dose manu eventureing a long-term cumulative influence (Adey, 1997 Galeev, 2000 Lai, 2005 Abu-Elsaoud, 2015). Modulated and pulsed radiofrequencies seem to be more impressive in producing effects (Belyaev, 2005 Lai, 2005). Low frequency modulations employ greater biological activi ty (Balmori, 2009).Microwave irradiation could affect ready growth, growth and seed germination (Hamada, 2007 Aladjadjiyan, 2010 Salama et al., 2011 (Scialabba and Tamburello, 2002 Monteiro et al., 2008 Ragha et al., 2011 Radzeviius et al., 2013 (Abu-Elsaoud, 2015). Low intensity microwave were reported non to affect the plant growth and development but the increased irradiation doses of microwave has decreased and slowed seed germination (Oprica, 2008). The direct effects of microwave on germination of food grains were studied by Ponomarev et al. (1996) where, a wavelength = 1 cm and irradiation exposure dose of up to 40 minutes were applied to barley, oats, and wheat seeds confidential information to improved germination rate with optimum effect after 20 minutes of microwave exposure (Ponomarev et al., 1996). A study of irradiating vegetable seeds with high power microwave radiations reported a stimulation influence of various germination and growth rate parameters by microwa ve (Radzeviius et al., 2013). The effect of microwave irradiation with a different power on various seed germination consequences of four different ornamental curb species has been studied by Aladjadjiyan (2002). The electroconductivity of leaf extract were monitored and increase in various germination consequences were observed (Aladjadjiyan, 2002). A comparative effect of microwave radiations on germination and growth of six different Egyptian ge nonypes were assessed utilise different exposure times, his data supported a dose dependent possible stimulation effect of microwave on growth and germination (Abu-Elsaoud, 2015). The response of barley seedlings to microwave radiations of 2.45 GHz after exposure to 0, 10, and 20 seconds of microwave radiations on four different genotypes (Creescu et al., 2013). Changes in peroxidase and catalase enzyme activities in Brassica napus were found to be dependent on microwave exposure time, seed condition and plant age (Oprica, 2008). The fr equencies of the cell plasm membrane vibrations of bio-objects lie in the mm-wave range, that range is thought to be essential to any living organism. Microwave irradiations induce resonant phenomena within biological system and have a stimulatory effect on biological organisms (Aladjadjiyan, 2002 Yanenko et al., 2004). Most microwave irradiation studies focused on possible biological effects from phone masts and microwave radiofrequencies on animal and human health (Santini et al., 2003 Hutter et al., 2006 Balmori, 2009). The biochemical mechanism by which microwave radiations affect biological systems of living organisms is not fully comprehended and the mechanism could vary according to the amplitude, frequency and the irradiation duty cycle (Monteiro et al., 2008 Aladjadjiyan, 2010). The present study was conducted to study the effect of seed irradiation with different doses of microwave radiations on the membrane electrolyte leakage, germination and growth of Egyptian barley Ho rdeum vulgare L seedlings.Materials and methodsPlant materialsSeeds of selected Barely Hordeum vulgare L. genotype Giza-129 were acquired from Agricultural Research Station at Ismailia, Agricultural Research Centre (ARC), Giza, Egypt in the months of November-December, 2016. The cereal lot of seeds was cleaned removing unwanted matter and stultificationd seeds.Radiofrequency irradiation treatmentMicrowave radiofrequency irradiation were carried out employ a magnetron with frequency of 2.45 GHz, wavelength of 12 cm, a maximum output power of 800 W, maximum intensity were estimated to be 51.5 kW.m3 by dividing the output power to the working volume m-3. Experimental details were presented in diagram (1). Seeds were scratch line soaked in distilled piss for 1 hour recommended by Aladjadjian and Svetleva (1997) to enhance the absorption of microwave energy. Seeds of selected barley genotype Hordeum vulgare cv. Giza-129 were divided into eight classs, each variant containing 30 seed s of (three replicas of ten seeds). The first group represent the untreated sustain and remaining seven variants were irradiated with different exposure periods to microwave (1, 5, 10, 30, 60, 300 and 600 seconds).Various germination traits were estimated and monitored during the experiment at different time-points 3, 5, 7, 9 and 12 days after sowing (DAS). Based on the obtained results, the percentage of germinated seeds Nk, germination rate Sk (seed.h-1), maximum fare of germinated seeds, relative germination coefficient (Wk) were calculated with the utilize germination formulas by Ciupak et al. (2007) presented in Table (1).Biomass and biomass allocationShoot and root biomass were determined for Triticum aestivum plants irradiated with 2.45 GHz radiofrequency and the untreated control. Biomass allocation within plants was calculated in g per g (S/R ratio, g.g-1) of total seedling biomass to avoid size of it effects, and calculated as a mean of three replicas. Data of Biomass allocation and shoot-to-root ratios were assessed statistically in plants irradiated with microwave radiofrequency versus the control ones to evaluate the change in biomass allocation pattern.Statistical analysesAnalysis of variance test (ANOVA) followed by Duncans multiple range comparisons were employed to analyse the results of barley after seed irradiation with microwave radiations. Further, correlational statistics and impartial linear regression analyses were in like manner performed using SPSS statistical software ver. 22 and Microsoft Excel share 2016 at a confidence level of 95%.ResultsSeed germinationThe influence of microwave radiations on various germination dynamics were assessed intensively on the first twelve days after seeds sowings (DAS) in Hordeum vulgare L. plant. Barley seeds were subjected to different exposure doses of microwave radiation from magnetron with 2450 MHz and 800 Watts. Germination indices monitored and assessed are number of germinated seeds (n k), percentage of germinated seeds (%), germination rate (Sk seed.h-1), germination index (GI), and the relative germination coefficient (Wk) at different time points 3, 5, 7, 9, and 12 DAS (days after seeds sowing). The percentages of germinated seeds were presented in Figure (1A-E) for different time points. A world-shattering change in the percentage of germinated seeds were observed after seed irradiation with microwave assessed by one-way abridgment of variance (ANOVA) followed by Duncans multiple range comparisons. Significant variations were observed versus the untreated control plant group. letter on figures 1 (A) to (E) represent the results of Duncans multiple range comparisons, where, different letter mean significant remainder (Figure 1). The maximum germination percentage observed were 100% recorded at MW dose of 5 seconds-5 DAS, 1, 5 seconds dose 7,9, 12 DAS. MW radiations observed to have a positive effect on germination at low doses of 1, and 5 seconds (Figure 1) these were assessed statistically by ANOVA and Duncans multiple comparisons.The general trend of MW radiations on seeds germination percentage was hearty negative and significant relationship (Figure 3A-E) revealed by both regression and Spearmans correlation i.e. increasing levels of MW radiations caused decrease in germination parameters especially high doses of MW.Other germination indices e.g. germination rate (Sk seed.h-1) were also recorded at different MW doses and time points (3, 5, 7, 9, 12). Germination rate in the untreated control 0.19 seed.h-1 5 and 7 days after seed sowing while in seeds treated with 1 and 5 seconds of MW the germination rate increased from 0.19 to 0.21 seed.h-1 revealing that not only the germination percentage increased but also the germination rate and speed (Figure 2A-D). Further, early germination was recorded after 1 and 5. Figures 3 (F-I) represent linear regression trend-line for the effect of MW radiation on germination rate, which had a stro ng inverse significant effect.Relative germination coefficient (Wk) were calculated and normalized to the control germination. Data of relative germination coefficient were presented in Figures (2E-H) at different time points (3, 5, 7,9) respectively. The relative germination coefficient increased after MW irradiation of 1 and 5 seconds (Figure 2E-H), while, Wk was decreased after irradiation with higher doses of MW radiations. Analysis of variance was carried out to assess the different surrounded by treatments control and were followed by Duncans multiple range comparisons. Linear regression trend-lines presented in figures (3K-N) represent the linear relationship between MW radiation doses and relative germination coefficient (Wk) after 3, 5, 7, 9 days after seed sowing. Inverse strong significant relationship between increasing doses of MW radiations and Wk. The germination index (GI) followed the same trend with increasing levels of microwave radiations (Figures 1, 3O).photosy nthetic pigments and GrowthShoot and root biomasses were estimated in Hordeum vulgare L. plants after irradiation to MW radiations. Shoot biomass ranged from 0.03 to 0.42 g/plant-FW where the maximum shoot fresh weight recorded after MW irradiation of 1 second dose and minimum in 600 seconds. MW radiations severely decreased the shoot biomass in barley (Figure, 4A). Root biomass, on the other hand, ranged from 0.03 to 36 g/plant-FW. The highest root fresh weight was recorded at 300 s MW irradiation dose. part minimum root fresh weight were recorded after 600 seconds MW dose (Figure 4B). Whole plant fresh weight ranged from 0.06 to 0.67 g/plant-FW. Shoot, root, and whole plant biomass showed a negative trend with increasing levels of MW radiations revealed by simple linear regression analysis and Spearmans correlation (Figure 7A,B,C). Microwave irradiation induced a significant decrease in shoot, root, and plant biomass in barley plants (Figure 7A,B,C).The demeanour or nutrient al location was assessed in monetary value of shoot and root biomass as shoot root ration (g.g-1) after seed irradiation with MW. Biomass behaviour was allocated toward barely shoot system after irradiation with 1 seconds of MW radiations. While, higher doses of MW induced nutrients to be allocated toward root system (figure 5B, 3O).Leaf chlorophyll contents increased significantly after 1 and 5 seconds of MW irradiations compared to the control (Figure 5A), however, MW doses from 300 and 600 seconds decreased significantly from the untreated control. Plant height was monitored after various MW irradiations doses and showed a significant decrease in response to MW (Figure 5) revealed by Duncans multiple range comparisons versus untreated control plants. Root volumes did not changes significantly with MW radiations except for the 600 seconds dose which showed a significant decrease versus control (Figure 5D)Membrane Ion leakage (%)Electrolyte leakage is a stress-induced injury that com monly used as a measure of plant response and permissiveness to stress (Bajji et al., 2002 Lee and Zhu, 2010). MW irradiation with dose 1 and 60 seconds did not induce a change in electrolyte leakage however, MW doses 5, 10, 30, 300 and 600 seconds significantly increased the electrolyte leakage compared to the untreated control (Figure 6). A strong negative significant relationship between increasing doses of MW radiations and electrolyte ion leakage (R2= 0.84 Pearson Correlation= -0.61 p-value DiscussionMicrowave irradiation with different exposure doses induced changes in various parameters of barley (H. vulgare genotype Giza-129). Germination parameters were dose-dependent and were stimulated by several exposure doses of microwave radiations. The percentage of germinated seeds, germination rate, relative germination coefficient and germination index at different time points were increased by short exposure to microwave radiations, however, height exposure doses of microwave-ind uced a significant decrease in germination consequences. Further, various growth parameters were increased by one or more low doses of microwave radiations and were significantly decreased by higher exposure doses. These results were found to be in agreement with (Abu-Elsaoud, 2015 Aladjadjiyan, 2002 Creescu et al., 2013 Ragha et al., 2011). Seed germination is completed with the protrusion of the radicle through the seed coat (Bewley Black, 1994). The subsequent seedling growth involves the establishment of the root and shoot systems. The hypocotyl growth is caused principally by cell expansion and/or by elongation. The low power 10.5 GHz irradiation reduces the rate and percentage of germination in daikon seeds and increases germination mean time, thus impairing seed germination. The germination reduction is linearly dependent on the MW power intensity incident on the seed. These findings support the simplified hypothesis that the power density on a plane perpendicular to wave di rection decreases with the inverse square of the distance from the source.Membrane electrolyte leakage accompanies the plant response to stresses were monitored at different microwave exposure doses. Electrolyte leakage is widely used as a measure of stress-induced injury in plants (Bajji et al., 2002 Lee and Zhu, 2010). According to our results microwave radiation with dose 1 seconds and 60 seconds did not induce a change in electrolyte leakage however, MW doses from 5 to 600 seconds significantly increased membrane electrolyte leakage compared to the untreated control. These results in agreement with previous results (Aladjadjiyan, 2002 Demidchik et al., 2014). A possible explanation by (Aladjadjiyan, 2002) suggests a hypothesis about the absorption of the microwave radiation energy by the hydrogen or magnesium atoms electrons in the chlorophyll molecule. The energy absorbed is redistributed and it causes changes in the chlorophyll molecule. By increasing the radiation power used for the treatment of the samples, the number of free ions in the extract decreases and hence its electroconductivity, too (Aladjadjiyan, 2002). Studies using patch-clamp method showed that the microwave exposure reduces trans-membrane protein channels opening in cultured chick myotubes probably because microwaves fire an alteration of intracellular enzymatic processes e.g. protein kinase activation (DInzeo et al., 1988) (DInzeo et al., 1988). In plant cells, the protein of water channels namely aquaporins of vacuolar membranes and plasm membranes are involved in the regulation of water movement dynamics in growth and development of plant cell and in stress responses (Maurel, 1997). In case of radish seedlings, microwaves may reduced water passage across cell membrane blocking aquaporins and do reduction of growth in a turgor-dependent manner (Scialabba and Tamburello, 2002).The increase of growth rate upon irradiation removal shows that during the elongation growth, the cell can partly repair damages occurred at the membrane level. There is a general consensus of opinion about the fact that MW induces a thermal detrimental effect over the biological system. In the present case, we assume that the damage induced by the low- power microwave exposure is non-thermal because a slight temperature increase (up to 25 oC over radish seeds has been demonstrated to induce germination and growth increase (Scialabba Melati, 1995). The reduced germination percentage and the delay seedling growth confirm the importance of a serious cause of concern about the influence of expo- sure to environmental MW fields. It can be stressed the importance of limiting in time the exposure to MW as suggested by the recovering ability of the biological system considered in the present research.Membrane Electrolyte leakage is an essential measure of the plants responses to various stresses. It is mostly associated with the K+ efflux, which is a common response in plant cells (Demidchik et al., 2014). The stress-induced electrolyte leakage is always accompanied by reactive oxygen species (ROS) generation and hence, leads to programmed cell death. Recent results exhibited that reactive oxygen species (ROS H2O2 and hydroxyl radicals) activates annexins, SKOR and GORK genes that catalyses K+ efflux from plant cells (Demidchik et al., 2014). Further, GORK-genes mediated potassium ion (K+) cause programmed cell death low oxidative stress. The intracellular endonucleases and proteases look to be blocked by potassium ions consequently, the efflux of these K+ stimulates these nucleases and proteases hydrolytic enzymes causing programmed cell death (PCD). Potassium ions could play a metabolic switch role under moderate stress conditions decreasing the anabolic reactions rate and stimulating catabolic reactions, leading to the release of energy compulsory for repairing and adaptation needs (Demidchik et al., 2014).The effect of microwaves on plants was the main purpose of the current study. Since it is a known problem, numerous other pieces of research were done on this topic. Having seen and observed other projects, we noticed that the major conflict was between whether microwaves affect plants germination or not. Our hypothesis was that they do affect it and, of course, it is well known that they do but it still made a challenge trying to prove it and it was found that every single trample affected the results. Since it is likely that other people who did similar projects have done some errors through their study, the results were not reliable and could not be considered accurate enough.