Effect of Different Microwave Doses on Vicia faba Germination, Growth and Genetic makeup Problem: Some areas are affected by high frequencies of microwaves; this affects plant growth, germination and genetic makeup, which leads to a loss in crop yield, including one of the major crops of high economic importance, the Vicia faba. Abstract: Plants and their stresses have been of very great importance lately. Since there are many kinds of stresses, we only studied one, but a very important one: microwaves and their effect on plants. Microwaves are going to be the focus of our study because of the drastic effects that they have, not only on humans and different mammals but on plants. In brief, we can say that microwaves are electromagnetic waves with wave length longer than those of terahertz (THz) frequencies, but relatively short for radio waves. There are lots of sites contaminated by microwave radiation from cellular radiation, power plants and high voltage towers, etc….. These affect the plants development, which drew our attention towards this project. Our hypothesis is that different doses of microwaves affect plant growth and development. To prove this hypothesis; we have to expose plants to different microwave doses, plant them in a green house under controlled conditions (soil, climate, humidity …) and observe them daily. Then we have to extract the DNA and RAPD of the DNA and determine the changes that occurred in their genes. Our conclusion from this study is that different microwave doses do affect plant germination. Introduction Plants and environmental stresses have been of most importance as a field of study and research through out the ages. It was found that the different stresses that plants are exposed to are outgoing and reduce the crop yield. Of these stresses, temperature, drought, salinity and microwaves are of major importance. Temperature, for example, is the principal factor determining vegetation zones. Global climate change caused by elevated greenhouse gasses was predicted to increase the proportion of hot, semi-arid land and the prevalence of high temperature and water stress. Oxidative stress from UV, photo-inhibition and pollutants such as troposphere ozone are associated problems. Drought is the main cause of low plant productivity and particularly of seasonally erratic productivity. Water is an increasingly continuous issue as there is not enough renewable fresh water at the right place and time to serve humans needs, regardless to say plants. Irrigation on demand and irrigation with poor quality water together lead to water logging and soil salinity so undermining the sustainability of the irrigated agriculture upon which food security depends. These reasons lead to research focusing on increasing knowledge of plant metabolic processes and their regulation. Developing novel protocols for the genetic manipulation of previously recalcitrant plant species is the most recent trend in research in regards to the solving of such problems. Microwaves are going to be the focus of our study because of the drastic effects that they have not only on humans and different mammals but on plants. In brief we can say that microwaves are electromagnetic waves with wave length longer than those of terahertz (THz) frequencies, but relatively short for radio waves. Microwaves have wavelength approximately in the range of 30 cm (frequency=1GHz) to 1mm (300 GHz). The microwave range includes ultra-high frequency (UHF) (0.303GHz), super high frequency (SHF) (3-30 GHz) and extremely high frequency (EHF) (30-300GHz) signals. Above 300 GHz, the absorption of electromagnetic radiation by earth atmosphere is so great that it is effectively opaque, until the atmosphere becomes transparent again in the so-called infrared and optical window frequency ranges. Microwaves were found to affect plants and plant growth severally. Long exposure to microwaves affects the germination of seeds or plants. However, weak microwaves don’t effect plant growth but as the microwave dose increases, the rate of germination decreases. Faba beans (Vicia faba) were found to be one of the most important sources of nutrition, being used in animal feed and -in some poor and developing countries- as a source of food. This very important crop is increasingly in demand for different uses as stated. It is noteworthy that the ability of using this crop and harvesting it in places were the environment has been contaminated with radiation will, of course, be of great benefit. In order to do so we need to properly identify the effect of microwaves on the genetic makeup of vicia faba and the correlation between that effect on both growth and germination of the beans. Aim: To Study the effect of microwaves in different doses on the germination, growth and genetic makeup of faba beans. Hypothesis: Seed exposure to microwaves will affect the germination, growth and genetic makeup of faba beans. Experiment Design: Giving the plant time for both germination and growth, we will correlate the relation between the exposure of different doses of microwaves and the plant germination, growth and genetic makeup. In studying that effect on plant germination we intend to observe the plant during stage of germination, taking in consideration parameters as time till germination, radical position and direction of growth and the time till shoots prevail. We put into consideration when studying plant growth the following parameters: plant height, time till complete growth, stem morphology, leaf morphology and the yield of the plants. Finally to correlate between that effect and the genetic makeup of the plants we will study the genetic makeup of the plants and the changes that occurred according to the different doses of microwave exposure using RAPD technique. Experiment variables 1. Microwave doses. 2. Planting in green house at the same conditions. 3. Parameters measured according to the breeder's program 4. Plant height, time till complete growth, Plant morphology and the yield of the plants Materials and Equipment:1. Microwave oven was used as a source of microwaves with frequency raging between 2.4-2.5 GHz. 2. Green house to plant the seeds to provide the same soil and irrigation method and climate 3. Triazol DNA extraction kit 4. PCR Machine 5. Electrophoresis unit 6. Gel imaging system 7. RAPD amplification kit Experimental Procedures: 1. Seeds were exposed to different microwave doses being studied namely, 2.4GHz, 2.45 GHz and 2.5GHz for a one minute interval 2. Upon exposure seeds were planted in a green house, in clay soil, being irrigated under controlled conditions with fixed climate and humidity 3. Daily observation of seeds germination for a period of five days 4. Daily observation of plant growth for a period of 14 days 5. Changes in plant growth were recorded 6. Pods were collected from the plants and then seeds were grinded 7. Total DNA extraction was done from the grinded seeds using the Triazol DNA extraction kit 8. RAPD of the extracted DNA using the RAPD Amplification kit. Results: After two weeks results were obtained, where we monitored different plant parameters visually for both germination and growth and using RAPD for the change in genetic makeup. The following results were tabulated. Plant Germination parameters Control Time till germination Direction of growth Time till shoots prevail Plant height after complete growth (cm) Time till complete growth (day) 2.40 2.45 2.50 1 day 2 day 3 day Under the soil Slightl y bent Under the soil Under Under the the soil soil Straig Straight ht Radical position Plant Growth parameters 1 day Microwave Dose (GHz) 3 days lateral 3 days 4 days 5 days Microwave Dose (GHz) Control 2.40 2.45 2.50 18 18 16 14 7 9 12 14 Stem morphology Normal Normal Normal Curved Leaf morphology Normal Normal Normal Smaller leaves Plant yield (pods) 7 6 6 4 1 2 3 Figure 1: Gel image of RAPD of the plants after being subjected to three different doses of microwaves 2.4 (1), 2.45 (2) and 2.5 (3). The arrow here indicates the molecular weight at which the new bands appeared proving the alteration of the genetic makeup of the faba beans upon exposure to different microwave doses. Conclusion: Different microwave doses do affect the germination, growth and genetic makeup of Vicia faba beans. Application: Under microwave stress, changes occur to the yield. Plants don't die but new bands appeared. Using these bands we can introduce new characters to plants that are present in regions that are contaminated with different microwave frequencies. When these bands become isolated, identified and sequenced we will use them to increase the resistance of plants that are cultivated in areas under microwave stress. Discussion To Study the effect of microwaves on plants was the main purpose of my study. Since it is a known problem, many other researches 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 step 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. However, at the end of our study the results showed that low microwave doses do not affect plant germination, meaning that as the microwave dose increases, the rate of germination decreases. References: http://en.wikipedia.org/wiki/Microwave http://en.wikipedia.org/wiki/Beans http://en.wikipedia.org/wiki/RAPD http://www.johnstonsarchive.net/environment/cellphone.html http://en.wikipedia.org/wiki/Global_warming