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Development of Bio-Machine Based on the Plant Response to External Stimuli

Development of Bio-Machine Based on the Plant Response to External Stimuli Hindawi Publishing Corporation Journal of Robotics Volume 2011, Article ID 124314, 7 pages doi:10.1155/2011/124314 Research Article Development of Bio-Machine Based on the Plant Response to External Stimuli 1, 2 1 2 K. Aditya, Ganesha Udupa, and Yongkwun Lee Department of Mechanical Engineering, Amrita Vishwa Vidyapeetham, Amritapuri Campus, Kollam 690525, India Centre for Bionics, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea Correspondence should be addressed to Yongkwun Lee, yklee@kist.re.kr Received 9 August 2011; Revised 31 October 2011; Accepted 11 November 2011 Academic Editor: Ivo Bukovsky Copyright © 2011 K. Aditya et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In the area of biorobotics, intense research work is being done based on plant intelligence. Any living cell continuously receives information from the environment. In this paper, research is conducted on the plant named descoingsii x haworthioides (Pepe) obtaining the action potential signals and its responses to stimulations of different light modes. The plant electrical signal is the reaction of plant’s stimulation owing to various environmental conditions. Action potentials are responsible for signaling between plant cells and communication from the plants can be achieved through modulation of various parameters of the electrical signal in the plant tissue. The modulated signals are used for providing information to the microcontroller’s algorithm for working of the bio-machine. The changes of frequency of action potentials in plant are studied. Electromyography (EMG) electrodes and needle- type conductive electrodes along with electronic modules are used to collect and transform the information from the plant. Inverse fast Fourier transform (IFFT) is used to convert signal in frequency domain into voltage signal for real-time analysis. The changes in frequency of the plant action potentials to different light modes are used for the control of the bio-machine. This work has paved the way for an extensive research towards plant intelligence. 1. Introduction elements. Because of that the resistance of different plants is not only ohmic but also frequency dependent. Much like humans, animals and plants have electrical signals In 1926, Bose used the isolated vascular bundles of a fern which pass through them, but plants do not have nerves (Blechnum nudum) to show that excitation was transmitted like humans and animals. Sanderson [1] was the first to as an electrical disturbance that appeared to be controlled by discover the action potentials (APs) in the stimulation of a similar physiological events as in animal nerves [8]. In 1968, Dionaea leaf. Hence, electrical signals do not belong only to Backster by using polygraph or “lie detector” can measure animal kingdom and humans [1]. Darwin [2] also found electrical resistance, and water would alter the resistance of the response of some carnivorous venus fly trap plants the leaf [9, 10]. Various studies have been carried out on the [2]. Generally in humans and animals when the muscle plant signal and its intelligence. Recently researchers found is voluntarily contracted, action appears. In plants it is that the transfer of volatile organic compounds (VOCs) found that action potentials are the signals caused by the signals among the plants. The signal is released by the emitter depolarization of plasma membrane [3, 4]. Green plants plant, and it is transported, absorbed, and perceived by the are able to show differentelectricalactivity, whichhas been receiver plant [11]. The fastest methods of long-distance known long time ago [5]. Moreover,exhaustivestudiesin this field began only in the last decades of the former communication between the plant tissues and the organs century along with the different contemporary experimental are bio-electrochemical or electrophysiological signals. The methods [6, 7]. Electrical phenomena in plants have com- effectiveness of such long-distance communication is clear, plicated character. Plant tissues are very complicated, highly since plants can respond to external stimuli (e.g., changes structured consisting of both conductive and insulative in temperature or osmotic environment, illumination level, 2 Journal of Robotics wounding, cutting, mechanical stimulation, or water avail- Needle-type electrode for wounding the plant leaf ability) and changes can be detected in the plant soon after the injury [7, 12]. The velocities of the propagation of electrical signals that have values from 0.5 mm to 4000 mm per second are sufficiently high to facilitate rapid long- distance communication, and these account for the rapid response phenomena observed in plants. The speed of propagation and the amplitude of action potential depend on the type of external stimulus [13]. It is also found that weak electrical signals of the chrysanthemum plant were tested by a touching test system of self-made double shields with platinum sensors [14]. Severalstudieshavereported Figure 1: Insertion of electrodes through the stomata (small pores; the effect of different stimuli that induce action potential the leaf is in dark green color) into the inner tissue. Plant electrical gradients in plants, mainly light/dark [15, 16]temperature signal behavior can be changed by this process. variations [17, 18], intense cold [19, 20], water availability [21, 22], mechanical wounding [23], and insects [22]. Also, it has been suggested that electrical signals could induce genetic programming [24, 25]. After the electrical signal is produced, it is transmitted through the plant to a specific organ or tissue, which generates immediate physiological actions in response to the stimulation [26]. Floranium lamp was developed to measure the voltage signal of the plant [27]. The Daisy team is working on the “The PLANTS” project that will enable the plant to control its own environment [28]. Ivanhenriques developed a Jurema action plant with electrodes clamped to the plant branches [29]. 1.1. Types of Signals in the Plant. Two types of signals in the plant have been described: fast signals (action potentials, APs) and slow signals (variation potentials, VPs). A new type of electrical potential signals, called system potentials, Figure 2: descoingsii x haworthioides plant for measuring the action has been postulated recently. The novel “system potentials” potentials. were detected in five different plant species, among them agricultural crops like tobacco (Nicotian atabacum), maize (Zea mays), Barley (Hordeum vulgare), and field bean (Vicia faba)[20]. Action potentials are an electrical waveform that The speed of propagation duration and amplitude of action is transmitted along the cell membrane [30], characterized potentials depend on the location of the working electrodes by a response. Plants respond to the environment change and the reference electrode. The changes in frequency in the according to its amplitude, frequency, and intensity [31, 32]. plant action potentials when it is exposed to different light Thus, action potentials allow cells, tissues, and organs to modes are observed and used to control the bio-machine. transmit electrical signals over short and long distances in plants. 2. Materials and Methods Variation potentials propagate in the plant, as temporal changes in the depolarization and repolarization of the cell 2.1. Plant and Laboratory Conditions. Different kinds of membrane; this kind of signal varies with the intensity of plants such as cactus (Cactaceae), soya beanplant (Glycine stimulation and appears to be associated with changes in max), chrysanthemum (Dendranthema x grandiflorum), and water tension or ion concentrations, creating a transient Pepe (descoingsii x haworthioides) are purchased from the electrochemical unbalance in the xylem [8, 33]. flower market and cultivated in the lab. It is observed that cactus is not responding and not showing any signal changes 1.2. Importance of Wounding Plant Leaf. If plant leaf is to the stimulus. The soybean and chrysanthemum plants are wounded, its action potential signal is stronger than that giving signals which are very weak to capture. The leaves of of unwounded leaf as shown in Figure 1 [20]. The strength these plants are very thin, and it is difficult to injure them of the inducing stimulus (wound signal) can influence using the needle electrodes. The leaves of the descoingsii x the frequency when compared to that of systematic signal haworthioides plant are very thick compared to the other (unwounded signal). plant leaves and sensitive to external stimulation. Hence In this research we are reporting a simple way for descoingsii x haworthioides is selected for further research. measuring the action potentials from the plant which is an It is well known that plants follow diurnal cycle [15]. We approach different from those applied by earlier researchers. assumed that plants are inactive during night time and rest Journal of Robotics 3 (a) (b) Figure 3: (a) Uni-Patch Tyco EMG electrodes (b) needle-type conductive electrodes. like human beings, and hence diurnal cycle is not considered during the experiment. Experiments are conducted during morning, afternoon, and evening, and it was found that there is no much variation in the results since the experiments are conducted in an air-conditioned room with almost similar environment conditions for all the light modes. In Figure 2 descoingsii x haworthioides plant is grown in a flower pot. The laboratory in which the experiments are carried out is a room with windows, and the temperature is kept at 26 C and the humidity at 75%. The light in the room is turned off at 23:00 hrs, creating total darkness. In the morning once again the plant is exposed to normal room conditions. The plant is watered every alternate days. Figure 4: Insertion of EMG electrode and needle-type conductive 2.2. Selection of Electrodes. Electromyography (EMG) elec- electrode (by wounding the leaf). trodes are generally used to detect the electric potentials generated by muscle cells when these cells are electrically or neurologically activated. Two kinds of electrodes are used for detecting the signals coming from the plant. LabVIEW is used for analyzing the signals coming from the First one is the Patch EMG electrode and the other plant. is conductive needle-type electrodes. Uni Patch Tyco EMG electrodes which are sensitive and circular in shape with 3.1.1. Description of the Circuit. Totally four EMG electrodes diameter of 20 mm and are used to collect the output signal and two needle-type conductive electrodes are used for from the plant leaves are shown in Figure 3(a). Another kind the experiment. Three leaves of descoingsii x haworthioides of electrodes used is needle-type conductive electrodes as plant are used in this experiment. Two leaves are used for shown in Figure 3(b), and it is made of copper. The length the input (sinusoidal) signal from the function generator of the electrode is 350 mm and the edge of the electrode is through two needle-type electrodes. The output signal is very sharp. The surface area of the leaf is nearly 2600 mm , collected through two EMG electrodes for detecting the and the area of electrode is nearly 314 mm . The ratio of action potential signals from the plant and stored in data leaf surface area to electrode area is nearly 8 : 1. Figure 4 acquisition system. The other two EMG electrodes are used shows the connection of these electrodes to the plant. EMG as reference electrodes connecting to each leaf. Figure 5 electrode is kept away at a distance of 250 mm from the shows the hierarchy of the system for experiment I which needle-type electrode to observe the action potentials. shows the block diagram of the system components. The plant receives an input signal from the function generator, 3. Experiments to Measure Electrical and the output signal from the plant is collected by using EMG electrodes and the data is stored in data acquisition Signal from the Plant Leaf system. The frequency of the function generator is chosen 3.1. Experiment I. The objective of this experiment is to based on the action potential signal strength coming from verify the behavior of action potentials in the plant. NI the plant leaf. Figure 6 shows the experimental setup. 4 Journal of Robotics Tracing of signal 1.5 from the plant leaves using 0.5 electrodes Data acquisition system (DAS) Pepe plant −0.5 −1 −1.5 −2 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1 Function generator NI LabVIEW Time (s) for input signal Output 1 Output 2 Figure 5: Hierarchy of system for experiment I. Input Figure 7: Output signals from the two leaves of the plant for a given sinusoidal input signal of same frequency. Input and output electrodes Reference electrodes Input signal Output signal Function generator Data acquisition system (DAS) Oscilloscope to Pepe plant Spike signal from measure the change electronic module in the action potentials Figure 6: Apparatus for measuring the action potentials from the plant leaves. Figure 8: Hierarchy of system for experiment II. and output electrode are shown, and the reference electrode 3.1.2. Results and Discussion. The experiments are carried connected to the ground is attached on the backside of the out with different frequencies ranging from 16 Khz to 24 Khz. leaf. The glowing of LED is shown by a rectangular mark. The output from the EMG electrode is connected to the NI- DAS (data acquisition system). The analog output signal is viewed on the computer screen, and the same is converted 3.2.1. Description of the Circuit. Differential circuit is con- to digital data and stored in the DAS. When the sinusoidal structed by using NE555 timer to generate the spike signals. signal frequency is at 22 Khz in the function generator, the Figure 10 shows the circuit designed to get the spike signal output signal from the plant has the same nature as that of as plant input signal so that change in the action potential input signal but with little variation in amplitude as shown signals from the plant leaf can be seen easily. These signals in Figure 7.In Figure 7, output signal from the two leaves are very sharp and assist in detecting small changes in the of the plant is shown for a given input signal of the same amplitude of output signals. The voltage of input signal frequency. This gives information about the output signal is 2.4 mV. The voltage of output signal collected from the from the plant for a given sinusoidal input signal. Other plant leaf is 1.2 mV. The input frequency is 23.4 Khz, and the types of signals such as square and triangle signals from the output frequency is 12.24 Khz. These readings are taken from function generator are also given as input signal to the plant the oscilloscope. leaf by varying the frequencies, but the output obtained for these types of signals are very weak, and there is no much 3.2.2. Procedure of the Experiment II. The input signal is variation in the amplitude. Hence sinusoidal signal is chosen given as spike signal from the electronic module shown in for this experiment. Figure 9, and the changes in action potential signals can be seen in oscilloscope. 3.2. Experiment II. Theobjective of thisexperimentisto The plant is exposed to three different light modes. verify the response of the plant when exposed to different The specification of the lamp used during experiments is light modes. Figure 8 shows the hierarchy of the system for GR2001 GRACE BIOLAMP, multifaceted reflector (MR16 experiment II. HALOGEN LAMP). The brightness of the lamp can be In this experiment only one leaf is used to measure the signal coming from the plant. One needle-type conductive adjusted. The maximum and minimum brightness of the electrode is used for giving input signal. Two EMG electrodes lamp ranges between 560 lumens and 710 lumens, and are used, one for collecting output signal and the other as medium brightness is 625 lumens. The mode of brightness a reference electrode. In Figure 9, only the input electrode of the table lamp is changed every 20 minutes. There is a Voltage (mV) Journal of Robotics 5 Oscilloscope Electronic module for the input signal Input and output electrodes LED lighting Figure 9: Apparatus for observing the change in action potential when plant is exposed to different light modes. 0 50 100 150 200 250 300 Output Input Time (s) 0.01 uF 100–1k Ohm No light Medium bright 2 3 Input TR Q Maximum bright DIS CC 5 6 R1 Figure 11: Frequency responses of the action potentials (APs) from THR CV R2 the plant leaf at different time intervals when exposed to three NE555 0.1 uF modes of light intensities. 0.01 uF Input (square signal) 3.3. Converting Frequency to Voltage. By using the inverse fast Fourier transform (IFFT) in MATLAB (R2010b), the fre- Output (spike signal quency values are converted to voltages as shown in Figure 13 as plant input signal) for the three different light modes. The differences in the Figure 10: Circuit design for the plant input signal as spike signal. amplitudes can be clearly seen at around 150 seconds. 4. Working and Control of Bio-Machine change in frequency in the plant signal every 5 to 10 sec The change in frequencies when plant is exposed to different during each measurement mode. Experiments are carried light conditions can be used as control signal for the out for each light intensity mode for about 300 seconds (no bio-machine as shown in Figure 14. Inverse fast Fourier light to maximum light brightness condition) after waiting transform (IFFT) can be used to show dominant amplitudes for 30–45 minutes to stabilize the lamp intensity. The change in the three different light modes. These values are stored in frequency of the action potential signal is observed from in microcontroller. Depending on the light modes, the bio- oscilloscope every 5 seconds using stop watch. machine will operate. Frequency counters can be used to see the response of plant to different light conditions, and the frequency can be converted into voltage signal by using the 3.2.3. Results and Discussion. The observed maximum fre- IC, LM2907. The direction change in the bio-machine can be quency values of the action potential signals in all the three seen by varying the light modes which results in the change modes of light are stored in the microcontroller. There is a lit- in frequency of the action potential signals from the plant. tle noise in the signal due to environmental vibrations. These effects are neglected since all the experiments are carried out in the similar environmental conditions. Figure 11 shows 5. Conclusion and Future Research frequency responses of the action potential signals from the plant leaf taking into account all the three modes of light In this research a simple method of detecting plant signals intensities. is investigated and the method is verified experimentally. There is an overlap in frequency values when there The change in frequency levels is observed when the plant is a change in light intensity from no light to maximum is exposed to different light conditions. Applying these brightness mode. These overlapped frequencies are removed results, bio-machine is constructed by designing a circuit during those particular time intervals by using the control and interfaced to a microcontroller. Software is written based algorithm developed for moving the bio-machine. The bio- on the developed algorithm to move the bio-machine in a machine will move to the right, left or, straight based on desired way. There is a little inconsistency in the movement the frequency values which are in close agreement with the of the bio-machine due to the random nature of the signals. values recorded in the microcontroller. Flow chart of the bio- However, the present work has paved the way for extensive machine movement is described in Figure 12. research on the plant intelligence in response to external Frequency (kHz) 6 Journal of Robotics Start Plant exposed to different light conditions Initialize and read the frequency value from the frequency counter Converting frequency to voltage Analog to digital conversion (ADC) Maximum bright Medium bright Check the Biomachine turns right Biomachine turns left frequency value Converting frequency to voltage Stop Figure 12: Flow chart for the movement of the bio-machine. −10 Figure 14: Prototype of bio-machine. −20 −30 250 300 150 200 50 100 also be used for the study of plant electrophysiology. Future Time (s) studies will be directed towards a better understanding of the plant action potential signals and its response to the No light other environmental conditions such as weather, presence Medium bright of sunlight, temperature, use of pesticides and test for the Maximum bright better movement of the bio-machine in response to these Figure 13: Real part of inverse fast Fourier transform (IFFT) for environmental conditions. converting the frequency signal to amplitude (mV). Acknowledgments stimuli as it holds out the true potential for innumerable The authors would like to thank the Korean Institute and very interesting applications. Green plants interfaced of Science and Technology (KIST) and Amrita Vishwa with a computer through data acquisition systems can be Vidyapeetham, Amrita School of Engineering, Amritapuri used as biosensors for monitoring the environment and to campus, for providing support to carry out the research and detect the effects of pollutants on the plants. This method can experiments. Amplitude (mV) Journal of Robotics 7 References [20] M. R. Zimmermann, H. Maischak, A. Mithofer ¨ , W. Boland, and H. H. Felle, “System potentials, a novel electrical long- [1] J. Burdon Sanderson, “Note on the electrical phenomena distance apoplastic signal in plants, induced by wounding,” which accompany irritation of the leaf of Dionaea muscipula,” Plant Physiology, vol. 149, no. 3, pp. 1593–1600, 2009. Royal Society of London Proceedings I, vol. 21, pp. 495–496, [21] J. Fromm and H. Fei, “Electrical signaling and gas exchange in maize plants of drying soil,” Plant Science, vol. 132, no. 2, pp. [2] C. 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Development of Bio-Machine Based on the Plant Response to External Stimuli

Aditya, K.; Udupa, Ganesha; Lee, Yongkwun
Journal of Robotics , Volume 2011 – Dec 20, 2011
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Hindawi Publishing Corporation
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Copyright © 2011 K. Aditya et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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1687-9600
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DOI
10.1155/2011/124314
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Abstract

Hindawi Publishing Corporation Journal of Robotics Volume 2011, Article ID 124314, 7 pages doi:10.1155/2011/124314 Research Article Development of Bio-Machine Based on the Plant Response to External Stimuli 1, 2 1 2 K. Aditya, Ganesha Udupa, and Yongkwun Lee Department of Mechanical Engineering, Amrita Vishwa Vidyapeetham, Amritapuri Campus, Kollam 690525, India Centre for Bionics, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea Correspondence should be addressed to Yongkwun Lee, yklee@kist.re.kr Received 9 August 2011; Revised 31 October 2011; Accepted 11 November 2011 Academic Editor: Ivo Bukovsky Copyright © 2011 K. Aditya et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In the area of biorobotics, intense research work is being done based on plant intelligence. Any living cell continuously receives information from the environment. In this paper, research is conducted on the plant named descoingsii x haworthioides (Pepe) obtaining the action potential signals and its responses to stimulations of different light modes. The plant electrical signal is the reaction of plant’s stimulation owing to various environmental conditions. Action potentials are responsible for signaling between plant cells and communication from the plants can be achieved through modulation of various parameters of the electrical signal in the plant tissue. The modulated signals are used for providing information to the microcontroller’s algorithm for working of the bio-machine. The changes of frequency of action potentials in plant are studied. Electromyography (EMG) electrodes and needle- type conductive electrodes along with electronic modules are used to collect and transform the information from the plant. Inverse fast Fourier transform (IFFT) is used to convert signal in frequency domain into voltage signal for real-time analysis. The changes in frequency of the plant action potentials to different light modes are used for the control of the bio-machine. This work has paved the way for an extensive research towards plant intelligence. 1. Introduction elements. Because of that the resistance of different plants is not only ohmic but also frequency dependent. Much like humans, animals and plants have electrical signals In 1926, Bose used the isolated vascular bundles of a fern which pass through them, but plants do not have nerves (Blechnum nudum) to show that excitation was transmitted like humans and animals. Sanderson [1] was the first to as an electrical disturbance that appeared to be controlled by discover the action potentials (APs) in the stimulation of a similar physiological events as in animal nerves [8]. In 1968, Dionaea leaf. Hence, electrical signals do not belong only to Backster by using polygraph or “lie detector” can measure animal kingdom and humans [1]. Darwin [2] also found electrical resistance, and water would alter the resistance of the response of some carnivorous venus fly trap plants the leaf [9, 10]. Various studies have been carried out on the [2]. Generally in humans and animals when the muscle plant signal and its intelligence. Recently researchers found is voluntarily contracted, action appears. In plants it is that the transfer of volatile organic compounds (VOCs) found that action potentials are the signals caused by the signals among the plants. The signal is released by the emitter depolarization of plasma membrane [3, 4]. Green plants plant, and it is transported, absorbed, and perceived by the are able to show differentelectricalactivity, whichhas been receiver plant [11]. The fastest methods of long-distance known long time ago [5]. Moreover,exhaustivestudiesin this field began only in the last decades of the former communication between the plant tissues and the organs century along with the different contemporary experimental are bio-electrochemical or electrophysiological signals. The methods [6, 7]. Electrical phenomena in plants have com- effectiveness of such long-distance communication is clear, plicated character. Plant tissues are very complicated, highly since plants can respond to external stimuli (e.g., changes structured consisting of both conductive and insulative in temperature or osmotic environment, illumination level, 2 Journal of Robotics wounding, cutting, mechanical stimulation, or water avail- Needle-type electrode for wounding the plant leaf ability) and changes can be detected in the plant soon after the injury [7, 12]. The velocities of the propagation of electrical signals that have values from 0.5 mm to 4000 mm per second are sufficiently high to facilitate rapid long- distance communication, and these account for the rapid response phenomena observed in plants. The speed of propagation and the amplitude of action potential depend on the type of external stimulus [13]. It is also found that weak electrical signals of the chrysanthemum plant were tested by a touching test system of self-made double shields with platinum sensors [14]. Severalstudieshavereported Figure 1: Insertion of electrodes through the stomata (small pores; the effect of different stimuli that induce action potential the leaf is in dark green color) into the inner tissue. Plant electrical gradients in plants, mainly light/dark [15, 16]temperature signal behavior can be changed by this process. variations [17, 18], intense cold [19, 20], water availability [21, 22], mechanical wounding [23], and insects [22]. Also, it has been suggested that electrical signals could induce genetic programming [24, 25]. After the electrical signal is produced, it is transmitted through the plant to a specific organ or tissue, which generates immediate physiological actions in response to the stimulation [26]. Floranium lamp was developed to measure the voltage signal of the plant [27]. The Daisy team is working on the “The PLANTS” project that will enable the plant to control its own environment [28]. Ivanhenriques developed a Jurema action plant with electrodes clamped to the plant branches [29]. 1.1. Types of Signals in the Plant. Two types of signals in the plant have been described: fast signals (action potentials, APs) and slow signals (variation potentials, VPs). A new type of electrical potential signals, called system potentials, Figure 2: descoingsii x haworthioides plant for measuring the action has been postulated recently. The novel “system potentials” potentials. were detected in five different plant species, among them agricultural crops like tobacco (Nicotian atabacum), maize (Zea mays), Barley (Hordeum vulgare), and field bean (Vicia faba)[20]. Action potentials are an electrical waveform that The speed of propagation duration and amplitude of action is transmitted along the cell membrane [30], characterized potentials depend on the location of the working electrodes by a response. Plants respond to the environment change and the reference electrode. The changes in frequency in the according to its amplitude, frequency, and intensity [31, 32]. plant action potentials when it is exposed to different light Thus, action potentials allow cells, tissues, and organs to modes are observed and used to control the bio-machine. transmit electrical signals over short and long distances in plants. 2. Materials and Methods Variation potentials propagate in the plant, as temporal changes in the depolarization and repolarization of the cell 2.1. Plant and Laboratory Conditions. Different kinds of membrane; this kind of signal varies with the intensity of plants such as cactus (Cactaceae), soya beanplant (Glycine stimulation and appears to be associated with changes in max), chrysanthemum (Dendranthema x grandiflorum), and water tension or ion concentrations, creating a transient Pepe (descoingsii x haworthioides) are purchased from the electrochemical unbalance in the xylem [8, 33]. flower market and cultivated in the lab. It is observed that cactus is not responding and not showing any signal changes 1.2. Importance of Wounding Plant Leaf. If plant leaf is to the stimulus. The soybean and chrysanthemum plants are wounded, its action potential signal is stronger than that giving signals which are very weak to capture. The leaves of of unwounded leaf as shown in Figure 1 [20]. The strength these plants are very thin, and it is difficult to injure them of the inducing stimulus (wound signal) can influence using the needle electrodes. The leaves of the descoingsii x the frequency when compared to that of systematic signal haworthioides plant are very thick compared to the other (unwounded signal). plant leaves and sensitive to external stimulation. Hence In this research we are reporting a simple way for descoingsii x haworthioides is selected for further research. measuring the action potentials from the plant which is an It is well known that plants follow diurnal cycle [15]. We approach different from those applied by earlier researchers. assumed that plants are inactive during night time and rest Journal of Robotics 3 (a) (b) Figure 3: (a) Uni-Patch Tyco EMG electrodes (b) needle-type conductive electrodes. like human beings, and hence diurnal cycle is not considered during the experiment. Experiments are conducted during morning, afternoon, and evening, and it was found that there is no much variation in the results since the experiments are conducted in an air-conditioned room with almost similar environment conditions for all the light modes. In Figure 2 descoingsii x haworthioides plant is grown in a flower pot. The laboratory in which the experiments are carried out is a room with windows, and the temperature is kept at 26 C and the humidity at 75%. The light in the room is turned off at 23:00 hrs, creating total darkness. In the morning once again the plant is exposed to normal room conditions. The plant is watered every alternate days. Figure 4: Insertion of EMG electrode and needle-type conductive 2.2. Selection of Electrodes. Electromyography (EMG) elec- electrode (by wounding the leaf). trodes are generally used to detect the electric potentials generated by muscle cells when these cells are electrically or neurologically activated. Two kinds of electrodes are used for detecting the signals coming from the plant. LabVIEW is used for analyzing the signals coming from the First one is the Patch EMG electrode and the other plant. is conductive needle-type electrodes. Uni Patch Tyco EMG electrodes which are sensitive and circular in shape with 3.1.1. Description of the Circuit. Totally four EMG electrodes diameter of 20 mm and are used to collect the output signal and two needle-type conductive electrodes are used for from the plant leaves are shown in Figure 3(a). Another kind the experiment. Three leaves of descoingsii x haworthioides of electrodes used is needle-type conductive electrodes as plant are used in this experiment. Two leaves are used for shown in Figure 3(b), and it is made of copper. The length the input (sinusoidal) signal from the function generator of the electrode is 350 mm and the edge of the electrode is through two needle-type electrodes. The output signal is very sharp. The surface area of the leaf is nearly 2600 mm , collected through two EMG electrodes for detecting the and the area of electrode is nearly 314 mm . The ratio of action potential signals from the plant and stored in data leaf surface area to electrode area is nearly 8 : 1. Figure 4 acquisition system. The other two EMG electrodes are used shows the connection of these electrodes to the plant. EMG as reference electrodes connecting to each leaf. Figure 5 electrode is kept away at a distance of 250 mm from the shows the hierarchy of the system for experiment I which needle-type electrode to observe the action potentials. shows the block diagram of the system components. The plant receives an input signal from the function generator, 3. Experiments to Measure Electrical and the output signal from the plant is collected by using EMG electrodes and the data is stored in data acquisition Signal from the Plant Leaf system. The frequency of the function generator is chosen 3.1. Experiment I. The objective of this experiment is to based on the action potential signal strength coming from verify the behavior of action potentials in the plant. NI the plant leaf. Figure 6 shows the experimental setup. 4 Journal of Robotics Tracing of signal 1.5 from the plant leaves using 0.5 electrodes Data acquisition system (DAS) Pepe plant −0.5 −1 −1.5 −2 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1 Function generator NI LabVIEW Time (s) for input signal Output 1 Output 2 Figure 5: Hierarchy of system for experiment I. Input Figure 7: Output signals from the two leaves of the plant for a given sinusoidal input signal of same frequency. Input and output electrodes Reference electrodes Input signal Output signal Function generator Data acquisition system (DAS) Oscilloscope to Pepe plant Spike signal from measure the change electronic module in the action potentials Figure 6: Apparatus for measuring the action potentials from the plant leaves. Figure 8: Hierarchy of system for experiment II. and output electrode are shown, and the reference electrode 3.1.2. Results and Discussion. The experiments are carried connected to the ground is attached on the backside of the out with different frequencies ranging from 16 Khz to 24 Khz. leaf. The glowing of LED is shown by a rectangular mark. The output from the EMG electrode is connected to the NI- DAS (data acquisition system). The analog output signal is viewed on the computer screen, and the same is converted 3.2.1. Description of the Circuit. Differential circuit is con- to digital data and stored in the DAS. When the sinusoidal structed by using NE555 timer to generate the spike signals. signal frequency is at 22 Khz in the function generator, the Figure 10 shows the circuit designed to get the spike signal output signal from the plant has the same nature as that of as plant input signal so that change in the action potential input signal but with little variation in amplitude as shown signals from the plant leaf can be seen easily. These signals in Figure 7.In Figure 7, output signal from the two leaves are very sharp and assist in detecting small changes in the of the plant is shown for a given input signal of the same amplitude of output signals. The voltage of input signal frequency. This gives information about the output signal is 2.4 mV. The voltage of output signal collected from the from the plant for a given sinusoidal input signal. Other plant leaf is 1.2 mV. The input frequency is 23.4 Khz, and the types of signals such as square and triangle signals from the output frequency is 12.24 Khz. These readings are taken from function generator are also given as input signal to the plant the oscilloscope. leaf by varying the frequencies, but the output obtained for these types of signals are very weak, and there is no much 3.2.2. Procedure of the Experiment II. The input signal is variation in the amplitude. Hence sinusoidal signal is chosen given as spike signal from the electronic module shown in for this experiment. Figure 9, and the changes in action potential signals can be seen in oscilloscope. 3.2. Experiment II. Theobjective of thisexperimentisto The plant is exposed to three different light modes. verify the response of the plant when exposed to different The specification of the lamp used during experiments is light modes. Figure 8 shows the hierarchy of the system for GR2001 GRACE BIOLAMP, multifaceted reflector (MR16 experiment II. HALOGEN LAMP). The brightness of the lamp can be In this experiment only one leaf is used to measure the signal coming from the plant. One needle-type conductive adjusted. The maximum and minimum brightness of the electrode is used for giving input signal. Two EMG electrodes lamp ranges between 560 lumens and 710 lumens, and are used, one for collecting output signal and the other as medium brightness is 625 lumens. The mode of brightness a reference electrode. In Figure 9, only the input electrode of the table lamp is changed every 20 minutes. There is a Voltage (mV) Journal of Robotics 5 Oscilloscope Electronic module for the input signal Input and output electrodes LED lighting Figure 9: Apparatus for observing the change in action potential when plant is exposed to different light modes. 0 50 100 150 200 250 300 Output Input Time (s) 0.01 uF 100–1k Ohm No light Medium bright 2 3 Input TR Q Maximum bright DIS CC 5 6 R1 Figure 11: Frequency responses of the action potentials (APs) from THR CV R2 the plant leaf at different time intervals when exposed to three NE555 0.1 uF modes of light intensities. 0.01 uF Input (square signal) 3.3. Converting Frequency to Voltage. By using the inverse fast Fourier transform (IFFT) in MATLAB (R2010b), the fre- Output (spike signal quency values are converted to voltages as shown in Figure 13 as plant input signal) for the three different light modes. The differences in the Figure 10: Circuit design for the plant input signal as spike signal. amplitudes can be clearly seen at around 150 seconds. 4. Working and Control of Bio-Machine change in frequency in the plant signal every 5 to 10 sec The change in frequencies when plant is exposed to different during each measurement mode. Experiments are carried light conditions can be used as control signal for the out for each light intensity mode for about 300 seconds (no bio-machine as shown in Figure 14. Inverse fast Fourier light to maximum light brightness condition) after waiting transform (IFFT) can be used to show dominant amplitudes for 30–45 minutes to stabilize the lamp intensity. The change in the three different light modes. These values are stored in frequency of the action potential signal is observed from in microcontroller. Depending on the light modes, the bio- oscilloscope every 5 seconds using stop watch. machine will operate. Frequency counters can be used to see the response of plant to different light conditions, and the frequency can be converted into voltage signal by using the 3.2.3. Results and Discussion. The observed maximum fre- IC, LM2907. The direction change in the bio-machine can be quency values of the action potential signals in all the three seen by varying the light modes which results in the change modes of light are stored in the microcontroller. There is a lit- in frequency of the action potential signals from the plant. tle noise in the signal due to environmental vibrations. These effects are neglected since all the experiments are carried out in the similar environmental conditions. Figure 11 shows 5. Conclusion and Future Research frequency responses of the action potential signals from the plant leaf taking into account all the three modes of light In this research a simple method of detecting plant signals intensities. is investigated and the method is verified experimentally. There is an overlap in frequency values when there The change in frequency levels is observed when the plant is a change in light intensity from no light to maximum is exposed to different light conditions. Applying these brightness mode. These overlapped frequencies are removed results, bio-machine is constructed by designing a circuit during those particular time intervals by using the control and interfaced to a microcontroller. Software is written based algorithm developed for moving the bio-machine. The bio- on the developed algorithm to move the bio-machine in a machine will move to the right, left or, straight based on desired way. There is a little inconsistency in the movement the frequency values which are in close agreement with the of the bio-machine due to the random nature of the signals. values recorded in the microcontroller. Flow chart of the bio- However, the present work has paved the way for extensive machine movement is described in Figure 12. research on the plant intelligence in response to external Frequency (kHz) 6 Journal of Robotics Start Plant exposed to different light conditions Initialize and read the frequency value from the frequency counter Converting frequency to voltage Analog to digital conversion (ADC) Maximum bright Medium bright Check the Biomachine turns right Biomachine turns left frequency value Converting frequency to voltage Stop Figure 12: Flow chart for the movement of the bio-machine. −10 Figure 14: Prototype of bio-machine. −20 −30 250 300 150 200 50 100 also be used for the study of plant electrophysiology. Future Time (s) studies will be directed towards a better understanding of the plant action potential signals and its response to the No light other environmental conditions such as weather, presence Medium bright of sunlight, temperature, use of pesticides and test for the Maximum bright better movement of the bio-machine in response to these Figure 13: Real part of inverse fast Fourier transform (IFFT) for environmental conditions. converting the frequency signal to amplitude (mV). Acknowledgments stimuli as it holds out the true potential for innumerable The authors would like to thank the Korean Institute and very interesting applications. Green plants interfaced of Science and Technology (KIST) and Amrita Vishwa with a computer through data acquisition systems can be Vidyapeetham, Amrita School of Engineering, Amritapuri used as biosensors for monitoring the environment and to campus, for providing support to carry out the research and detect the effects of pollutants on the plants. This method can experiments. Amplitude (mV) Journal of Robotics 7 References [20] M. R. Zimmermann, H. Maischak, A. Mithofer ¨ , W. Boland, and H. H. Felle, “System potentials, a novel electrical long- [1] J. Burdon Sanderson, “Note on the electrical phenomena distance apoplastic signal in plants, induced by wounding,” which accompany irritation of the leaf of Dionaea muscipula,” Plant Physiology, vol. 149, no. 3, pp. 1593–1600, 2009. Royal Society of London Proceedings I, vol. 21, pp. 495–496, [21] J. Fromm and H. Fei, “Electrical signaling and gas exchange in maize plants of drying soil,” Plant Science, vol. 132, no. 2, pp. [2] C. 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