Name: enhancement of the initial growth rate of agricultural plants by using static magnetic fields
Uploaded: 2016-07-08T13:00:00.000+00:00
Duration: 5 min 17 s
Description: Watch this Scientific Journal Video about Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields at JoVE.com

This study received supported from the National Research Foundation of Korea (NRF) (2011-0012728). A poster presenting this study was awarded the Best Poster Award by the Korean Society of Applied Biological Sciences (KSABC).

1. Ajustes iniciales <ol><li> Especies de plantas agrícolas <ol><li> Uso del jardín Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), y Mescluns (Lepidium sativum) semillas. NOTA: Impatiens balsamina (bálsamo del jardín o Rose Balsam) es una especie nativa de la India; algunos miembros también se encuentran en Myanmar. Komatsuna (Brassica rapa var. Perviridis o komatsuna) es una variante de la misma especie que el nabo común. El mastuerzo (Lepidium sativum) es un tipo de hierba que se taxonómicamente relacionado con el berro y la mostaza. Tienen sabores y aromas similares, para los que se utilizan comercialmente 5,7. </li></ol></li><li> Los cultivos de plantas <ol><li> Cultura Jardín Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), y mezclums (Lepidium sativum) semillas en un 100 mm de diámetro (100 pi) placa de Petri. Asegúrese de que una placa contiene sólo un tipo de especie. </li><li> Para las condiciones de cultivo, colocar las semillas en una toalla de celulosa. Sumergir la toalla y semillas en agua destilada tres veces. Medir y confirmar que el laboratorio de interior RT es de 18-25 ° C, con una humedad que oscila de 65-75% (por favor, compruebe la sección 3.1.2). </li><li> Por número de semillas, cultivo de 10 ± 1 semillas de bálsamo del jardín, 50 ± 10 semillas de Mizuna, 330 ± 20 semillas de Komatsuna, y 380 ± 20 semillas de Mescluns. Use condiciones idénticas medidas como 18-25 ° C, con una humedad que oscila de 65-75% (por favor, compruebe la sección 2.1.1). NOTA: Todos los experimentos se realizaron en condiciones de interior con la humedad regulado y rango de temperatura en el laboratorio. La humedad y la temperatura no era estática, pero siempre y cuando las condiciones idénticas para tratar el imán y el grupo control. </li></ol></li></ol> 2. Cultura de cuatro plantas agrícolas<ol><li> Procedimiento experimental <ol><li> Siga la Sección 1.2.3) para las especies de plantas y las condiciones de cultivo en los grupos control y aplicado imán. </li><li> Aplicar tres imanes de 1750 ± 350 Gauss (10.000 Gauss = 1 Tesla) en la parte inferior de las placas de 100 pi de Garden Balsam. Durante la aplicación, asegúrese de que los tres imanes no están en contacto directo con las semillas, y se separan por la parte inferior de plástico de la placa de Petri. La distancia directa entre las semillas y los imanes debe ser de 2-4 mm. Aplicar los imanes de 168 horas (7 días) durante cuatro plantas agrícolas. </li><li> Siguiendo todos los pasos de forma idéntica en 2.1.2), se aplican dos imanes, uno (N orientados hacia los lados hacia arriba) en el lado S imán superior y otra (hacia arriba) en la parte inferior de la placa de cultivo de bálsamo del jardín. NOTA: Los postes se aplican de manera diferente en Garden Balsam. Sin embargo, la orientación de polos no se considera como un factor crucial en este estudio para alteraciones del crecimiento, como todos los entornos son idénticos excepto por la direccióndel flujo magnético. El propósito de N y S aplicación polos para bálsamo del jardín era para ver su capacidad práctica en su uso en los campos, donde la orientación de polos podría ser difícil de manejar. </li></ol></li></ol> 3. La tinción de tubulina Jardín Balsam <ol><li> Aplicaciones de imán con Regulado Condición Luz <ol><li> Coloque dos imanes (polo N hacia arriba) parte inferior de la placa de 100 mm durante 48 horas, utilizando las condiciones en el paso 1.2.2. NOTA: Para la modificación de la luz, las placas de cultivo fueron colocados en un estante de plástico en la incubadora. Incubadora se utilizó para la intercepción de la luz y la temperatura se mantiene a 25 ° C durante 48 horas en ambientes oscuros. Con el tiempo, esta condición no se utilizó en este experimento debido a las altas variaciones en la longitud de crecimiento. </li></ol></li><li> La tinción de plantas <ol><li> Fijar la totalidad de impatiens SPP planta de flor doble, (incluyendo el tronco y las raíces) crecido en idénticas condiciones con la etapa 3.1.2) en 4% de paraformaldehído y 0,1 Mtampón de fosfato (pH 7,4) durante 15 min. </li><li> Retirar la muestra impatiens y sumergirse durante 2 horas en tampón de bloqueo (2% de suero de caballo / 1% de albúmina de suero bovino / 0,1% de Triton X-100 en PBS, pH 7,5). Lavar la muestra impatiens por inmersión con PBS durante 15 min. </li><li> Para inmunotinción doble, incubar la muestra con el anticuerpo primario, anti-tubulina alfa (1: 1.000), O / N a 4 ° C. </li><li> Retire la muestra y sumergir las muestras una vez con PBS durante 10 min para lavar. Uso conjugado con FITC anti-IgG de ratón (1: 400) como el anticuerpo secundario e incubar durante 2 horas a 25 ° C. </li><li> Sumergir la muestra en PBS y de cubre toda la muestra en la parte inferior de la placa de 24 pocillos. Obtención de imágenes mediante un microscopio de fluorescencia convencional para observar la orientación de la tubulina (λ = 550 nm, magnificado a 100X, 200X y 400X). NOTA: En este caso, el grupo tratado imán (n = 10) y controles (n = 11) se confirmaron para Garden Balsam (Impatiens balsamina) solamente, que se cultiva en condiciones de no-oscuros. </li&gt; </ol></li></ol> 4. Métodos de Recogida de Datos <ol><li> Lapso de tiempo Creación de crecimiento de las plantas Cuatro Agrícola <ol><li> Fotografía de la planta a intervalos de 10 min, mediante el establecimiento de obturador para auto (esto se puede hacer en cualquier cámara digital). Ajuste la abertura a F 3.2 y el valor ISO 400. </li><li> Recoger 700-900 imágenes durante 7-10 días. Conectar la cámara con los cables eléctricos ya batería puede estar agotada. </li><li> Arrastre las imágenes haciendo clic y colocar cada imagen cronológicamente bajo la línea de transmisión de creación de películas con el software (ver materiales y equipos de mesa). Póngalo en una línea de streaming en duraciones iguales de 0,045 a 0,05 seg para cada película en un total de 30-40 seg. La hora de llegada a fin de que no haya espacios oscuros con la selección de la cada imagen en un orden cronológico. </li><li> Después del paso 4.1.3, haga clic en el botón de reproducción en el software para asegurar la película compilado en un lapso de tiempo de diapositivas de vídeo de 30-40 segundos y haga clic en Procesar y guardar en formato .avi o .mpeg. Para el tamaño de markers, utilice cuartos de Canadá, un penique americano, y una regla de centímetros en el lado de la foto. </li><li> Realizar la prueba t y el diagrama de caja para el análisis estadístico 11,12. NOTA: Los grupos de resúmenes de cinco números se utilizaron para calcular el límite inferior (L) valor como Q1 - [1,5 × (Q3 - Q1)] y el límite superior (H) valor como Q3 + [1.5 × (Q3 - Q1) ]. Este enfoque se incorporó en el paso 1.2.2 para la recolección de datos de longitud 11. Los valores de L y H muestran el área 99% de las parcelas T-distribución, lo que significa que los puntos de datos observados fuera de este rango pueden ser considerados valores atípicos. Los diagramas de caja y la t de Student-test se utilizaron para analizar las diferencias en las alturas de las plántulas 12. </li></ol></li></ol>

<ol>
	<li>Martin, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+measurement+of+pollen+tube+growth+inhibition.">In vitro measurement of pollen tube growth inhibition.</a> Plant Physiol. 49, 924-925 (1972).</li><li>Pfahler, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+germination+characteristics+of+maize+pollen+to+detect+biological+activity+of+environmental+pollutants.">In vitro germination characteristics of maize pollen to detect biological activity of environmental pollutants.</a> Environ Health Perspect. 37, 125-132 (1981).</li><li>Yao, Z., Tan, X., Du, H., Luo, B., Liu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-current+microwave+ion+source+with+permanent+magnet+and+its+beam+emittance+measurement.">A high-current microwave ion source with permanent magnet and its beam emittance measurement.</a> Rev Sci Instrum. 79, 073304 (2008).</li><li>Hendrickson, C. L., Drader, J. J., Laude, D. A., Guan, S., Marshall, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fourier+transform+ion+cyclotron+resonance+mass+spectrometry+in+a+20+T+resistive+magnet.">Fourier transform ion cyclotron resonance mass spectrometry in a 20 T resistive magnet.</a> Rapid Commun Mass Spectrom. 10, 1829-1832 (1996).</li><li>Namba, K., Sasao, A., Shibusawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EFFECT+OF+MAGNETIC+FIELD+ON+GERMINATION+AND+PLANT+GROWTH.">EFFECT OF MAGNETIC FIELD ON GERMINATION AND PLANT GROWTH.</a> Acta Hort. 399, 143-148 (1995).</li><li>Hirota, N., Nakagawa, J., Kitazawa, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+a+magnetic+field+on+the+germination+of+plants.">Effects of a magnetic field on the germination of plants.</a> Journals of Applied Physics. 85, 5717-5719 (1999).</li><li>Penuelas, J., Llusia, J., Martinez, B., Fontcuberta, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diamagnetic+Susceptibility+and+Root+Growth+Responses+to+Magnetic+Fields+in+Lens+culinaris,+Glycine+soja,+and+Triticum+aestivum.">Diamagnetic Susceptibility and Root Growth Responses to Magnetic Fields in Lens culinaris, Glycine soja, and Triticum aestivum.</a> Electromagnetic Biology and Medicine. 23, 97-112 (2004).</li><li>Carbonell, M. V., Martinez, E., Amaya, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+germination+in+rice+(Oryza+Sativa+L.)+by+a+static+magnetic+field.">Stimulation of germination in rice (Oryza Sativa L.) by a static magnetic field.</a> Electro- and Magnetobiology. 19, 121-128 (2000).</li><li>Oakley, R. V., Wang, Y. S., Ramakrishna, W., Harding, S. A., Tsai, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+expansion+and+expression+of+alpha-+and+beta-tubulin+gene+families+in+Populus.">Differential expansion and expression of alpha- and beta-tubulin gene families in Populus.</a> Plant Physiol. 145, 961-973 (2007).</li><li>Hoson, T., Matsumoto, S., Soga, K., Wakabayashi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+microtubules+are+responsible+for+gravity+resistance+in+plants.">Cortical microtubules are responsible for gravity resistance in plants.</a> Plant Signal Behav. 5, 752-754 (2010).</li><li>Kim, S., Im, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Static+magnetic+fields+inhibit+proliferation+and+disperse+subcellular+localization+of+gamma+complex+protein3+in+cultured+C2C12+myoblast+cells.">Static magnetic fields inhibit proliferation and disperse subcellular localization of gamma complex protein3 in cultured C2C12 myoblast cells.</a> Cell Biochem Biophys. 57, 1-8 (2010).</li><li>Benjamini, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opening+the+Box+of+a+Boxplot.">Opening the Box of a Boxplot.</a> The American Statistician. 42, 257-262 (1988).</li></ol>

The Authors have nothing to disclose.

En todas las condiciones, los imanes deben aplicarse bajo la placa de Petri. Este estudio examinó la influencia de los campos magnéticos sobre la tasa de crecimiento de las semillas de varias especies agrícolas, con especial atención en bálsamo del jardín como representante de plantas agrícolas. Por ejemplo, la tinción de tubulina se realizó sobre el Garden Balsam para evaluar los cambios en el nivel molecular en la raíz y el tallo micro-estructuras esqueléticas que sugiere la influencia del campo magnético ...

La germinación es el crecimiento de una planta que da lugar a la formación de la planta de semillero 1. Bajo ciertas condiciones, la germinación de semillas comienza y los tejidos embrionarios reanudar el crecimiento. Comienza con la hidratación de la semilla con el fin de activar las enzimas para la germinación. Las semillas pueden ser inducidos a germinar in vitro (en una placa de Petri o tubo de ensayo) 1,2. Los campos magnéticos estáticos son fuerzas especiales que provocan movimientos de las moléculas con cargas iónicas por medio de la fuerza de Lorentz 3,4. se forma una fuerza de Lorentz cuando ionizados o cargados objeto se mueve bajo un campo magnético. Cada material se formó con los átomos que están compuestos de electrones y protones. Cuando los campos magnéticos se hacen presentes, ya sea estática o alterna, que afecta a la circulación de material cargado. Esto también se aplica a las plantas y las moléculas de agua, que afecta a la condición molécula intracelular. En un estudio anterior, se utilizaron bobinas electromagnéticaspara generar campos magnéticos pulsados, y las plantas &#39;komatsuna&#39; fueron elegidos como los temas 5. En el presente estudio, el imán genera campos magnéticos estáticos se utilizan para dar un efecto similar pero diferente como un estudio de la expansión de la fuerza de Lorentz. La frecuencia del campo magnético, en lugar de su polaridad, es un factor crucial para la germinación de las plantas. Estudios anteriores han sugerido que las tasas de germinación máximas fueron 20% más alto que el control cuando la frecuencia del campo magnético fue de aproximadamente 10 Hz. Cuando el campo se quita de forma retrógrada, la tasa de crecimiento se deteriora 5. Los campos magnéticos estáticos tienen un efecto considerable en el 6-8 inicial de crecimiento, principalmente en la germinación y crecimiento de las raíces 6 7. En el presente estudio, hemos utilizado los imanes estáticos para examinar la posibilidad de regular el crecimiento de plantas agrícolas mediante el uso de campos magnéticos. En particular, el objetivo fue determine si ciertas condiciones de aplicación del campo magnético podría aumentar las tasas de crecimiento a niveles más altos que los mencionados en la literatura. Además, si la germinación inicial de las plantas se puede incrementar con éxito utilizando un campo magnético, el uso de fertilizantes químicos se puede evitar.

<table><tbody><tr><td>Static magnets</td><td>JIM</td><td></td><td>2000Gauss</td></tr><tr><td>2% horse serum/1% bovine serum albumin/0.1% Triton X-100</td><td>Sigma-Aldrich</td><td>Merged with 55514</td><td>Blocking buffer</td></tr><tr><td>Primary antibody</td><td>Santa Cruz Biotechnology</td><td>sc-8035</td><td>a-Tubulin</td></tr><tr><td>Secondary antibody</td><td>Santa Cruz Biotechnology</td><td>sc-2010</td><td>FITC-conjugated anti-mouse IgG</td></tr><tr><td>time lapse photographic techniques</td><td>Manually controlled</td><td></td><td>ISO value 400 &amp;amp; aperture F 3.2</td></tr><tr><td>Sony Vegas Pro 13.0</td><td>Sony</td><td></td><td></td></tr></tbody></table>

enhancement of the initial growth rate of agricultural plants by using static magnetic fields

dispositivos electrónicos y cables de alta tensión inducen campos magnéticos. Un campo magnético de Gauss 1,300-2,500 (0.2 Tesla) se aplicó a placas de Petri que contienen semillas de bálsamo del jardín (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), y Mescluns (Lepidium sativum ). Se aplicaron los imanes bajo la placa de cultivo. Durante los 4 días de la aplicación, se observó que la longitud del tallo y la raíz aumentado. El grupo sometido a un tratamiento de campo magnético (n = 10) mostró un 1,4 veces más rápido ritmo de crecimiento en comparación con el grupo control (n = 11) en un total de 8 días (p &lt;0,0005). Esta tasa es 20% superior a la reportada en estudios previos. Las líneas complejas de tubulina no tienen puntos de conexión, pero los puntos de conexión se producen de la aplicación de imanes. Esto muestra completa diferencia del control, lo que significa arreglos anormales. Sin embargo, la causa exacta no está clara. estas resULTS de mejora del crecimiento de la aplicación de imanes sugieren que es posible mejorar la tasa de crecimiento, aumentar la productividad, o controlar la velocidad de germinación de las plantas mediante la aplicación de campos magnéticos estáticos. Además, los campos magnéticos pueden causar cambios fisiológicos en las células vegetales y pueden inducir el crecimiento. Por lo tanto, la estimulación con un campo magnético puede tener posibles efectos que son similares a las de los fertilizantes químicos, lo que significa que el uso de fertilizantes puede ser evitado.

Tinción de tubulina mostró dispersa o estructuras adelgazado en las plantas cultivadas en presencia del imán en comparación con el control (Figura 2). Por otra parte, los estudios de 7 días de lapso de tiempo con las plantas agrícolas, incluidas Komatsuna (Brassica rapa var. Perviridis) y Mescluns (Lepidium sativum) indicaron que un imán campo magnético estático deriva aumenta el crecimiento inicial de estas plantas (Figura 3)....

Watch this Scientific Journal Video about Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields at JoVE.com

Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields

The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.

Mejora de la tasa de crecimiento inicial de las plantas agrícolas mediante el uso de campos magnéticos estáticos

Electronic devices and high-voltage wires induce magnetic fields. A magnetic field of 1,300-2,500 Gauss (0.2 Tesla) was applied to Petri dishes containing ...

enhancement-initial-growth-rate-agricultural-plants-using-static

Research

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Unit 1

Magnetic Forces and Fields

SCIENTISTS IN ACTION

13.2K vistas.  Yonsei University. The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.

Video: Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields

Geomagnetic Field (Gmf) and Plant Evolution: Investigating the Effects of Gmf Reversal on Arabidopsis thaliana Development and Gene Expression

One of the most stimulating observations in plant evolution is a correlation between the occurrence of geomagnetic field (GMF) reversals (or excursions) and the moment of the radiation of Angiosperms. This led to the hypothesis that alterations in GMF polarity may play a role in plant evolution. Here, we describe a method to test this hypothesis by exposing Arabidopsis thaliana to artificially reversed GMF conditions. We used a three-axis magnetometer and the collected data were used to calculate the magnitude of the GMF. Three DC power supplies were connected to three Helmholtz coil pairs and were controlled by a computer to alter the GMF conditions. Plants grown in Petri plates were exposed to both normal and reversed GMF conditions. Sham exposure experiments were also performed. Exposed plants were photographed during the experiment and images were analyzed to calculate root length and leaf areas. Arabidopsis total RNA was extracted and Quantitative Real Time-PCR (qPCR) analyses were performed on gene expression of CRUCIFERIN 3 (CRU3), copper transport protein1 (COTP1), Redox Responsive Transcription Factor1 (RRTF1), Fe Superoxide Dismutase 1, (FSD1), Catalase3 (CAT3), Thylakoidal Ascorbate Peroxidase (TAPX), a cytosolic Ascorbate Peroxidase1 (APX1), and NADPH/respiratory burst oxidase protein D (RbohD). Four different reference genes were analysed to normalize the results of the qPCR. The best of the four genes was selected and the most stable gene for normalization was used. Our data show for the first time that reversing the GMF polarity using triaxial coils has significant effects on plant growth and gene expression. This supports the hypothesis that GMF reversal contributes to inducing changes in plant development that might justify a higher selective pressure, eventually leading to plant evolution.

Geomagnetic Field Gmf and Plant Evolution: Investigating the Effects of Gmf Reversal on Arabidopsis thaliana Development and Gene Expression

This protocol describes a method to test the hypothesis that the reversal of the effective geomagnetic field (GMF) induces differential gene expression and alters the morphology of Arabidopsis thaliana. A triaxial octagonal system of Helmholtz coil-pairs was used to artificially generate reversed GMF conditions in the plant growth chamber.

Campo geomagnético (Gmf) y evolución de las plantas: La investigación de los efectos de Gmf Reversión en<em&gt; Arabidopsis thaliana</em&gt; Desarrollo y Expresión Génica

One of the most stimulating observations in plant evolution is a correlation between the occurrence of geomagnetic field (GMF) reversals (or excursions) ...

Investigación

Biología del Desarrollo

Robotic Sensing and Stimuli Provision for Guided Plant Growth

Robot systems are actively researched for manipulation of natural plants, typically restricted to agricultural automation activities such as harvest, irrigation, and mechanical weed control. Extending this research, we introduce here a novel methodology to manipulate the directional growth of plants via their natural mechanisms for signaling and hormone distribution. An effective methodology of robotic stimuli provision can open up possibilities for new experimentation with later developmental phases in plants, or for new biotechnology applications such as shaping plants for green walls. Interaction with plants presents several robotic challenges, including short-range sensing of small and variable plant organs, and the controlled actuation of plant responses that are impacted by the environment in addition to the provided stimuli. In order to steer plant growth, we develop a group of immobile robots with sensors to detect the proximity of growing tips, and with diodes to provide light stimuli that actuate phototropism. The robots are tested with the climbing common bean, Phaseolus vulgaris, in experiments having durations up to five weeks in a controlled environment. With robots sequentially emitting blue light-peak emission at wavelength 465 nm-plant growth is successfully steered through successive binary decisions along mechanical supports to reach target positions. Growth patterns are tested in a setup up to 180 cm in height, with plant stems grown up to roughly 250 cm in cumulative length over a period of approximately seven weeks. The robots coordinate themselves and operate fully autonomously. They detect approaching plant tips by infrared proximity sensors and communicate via radio to switch between blue light stimuli and dormant status, as required. Overall, the obtained results support the effectiveness of combining robot and plant experiment methodologies, for the study of potentially complex interactions between natural and engineered autonomous systems.

Distributed robot nodes provide sequences of blue light stimuli to steer the growth trajectories of climbing plants. By activating natural phototropism, the robots guide the plants through binary left-right decisions, growing them into predefined patterns that by contrast are not possible when the robots are dormant.

Suministro de sensistes y de sinensistes para el crecimiento guiado de la planta

Robot systems are actively researched for manipulation of natural plants, typically restricted to agricultural automation activities such as harvest, ...

Ingeniería

Control and Disposal of Invasive Japanese Knotweed Reynoutria japonica Houtt. Using Microwave Treatment

The study aims to assess the effectiveness of microwave treatment (MWT) at a frequency of 2.45 GHz and a power of 800 W to control Japanese knotweed (Reynoutria japonica Houtt.) using a self-propelled device that was built in the in-house facility. The MWT was applied in the field population of knotweed in July 2022. First, plants were mechanically moved from the area of 1 m2, and next, the cut shoots around 4 cm high were microwave-treated for 25 min, 20 min, and 15 min. The control treatments were: 1) only cut plants and 2) rhizomes dug out to 30 cm deep. The effectiveness of the microwave treatments was observed for the next 11 months by counting the number of newly grown shoots. The results showed that a 25 min MWT was 100% effective in Japanese knotweed loss of vitality, while a 15 min MWT microwave treatment stimulated plant growth by around 50%, compared to controls. Rhizomes were dug out in a separate in vitro experiment for laboratory testing. The rhizomes were categorized by thickness and subjected to a 60 s MWT using a commercial microwave, after which their temperature and vitality were assessed. The temperature of rhizomes following MWT depended on their thickness. Those rhizomes that warmed to temperatures above 42 &#176;C were effectively destroyed. Summing up, the time plants are exposed to microwaves plays a major role in the effectiveness of this method. The longer the exposure to MWT, the better control. The thinner the rhizomes, the more effective the in vitro MWT rhizomes disposal.

This protocol provides a method for microwave control of Japanese knotweed in the field and disposal of dug-up rhizomes in laboratory conditions. The advantages and disadvantages of both methods are discussed. Future research directions are also suggested to optimize the use of microwaves for controlling Japanese knotweed.

The study aims to assess the effectiveness of microwave treatment (MWT) at a frequency of 2.45 GHz and a power of 800 W to control Japanese knotweed ...

Medio ambiente

3D Magnetic Stem Cell Aggregation and Bioreactor Maturation for Cartilage Regeneration

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach recently explored for the development of autologous replacements involves the differentiation of stem cells into chondrocytes. To initiate this chondrogenesis, a degree of compaction of the stem cells is required; hence, we demonstrated the feasibility of magnetically condensing cells, both within thick scaffolds and scaffold-free, using miniaturized magnetic field sources as cell attractors. This magnetic approach was also used to guide aggregate fusion and to build scaffold-free, organized, three-dimensional (3D) tissues several millimeters in size. In addition to having an enhanced size, the tissue formed by magnetic-driven fusion presented a significant increase in the expression of collagen II, and a similar trend was observed for aggrecan expression. As the native cartilage was subjected to forces that influenced its 3D structure, dynamic maturation was also performed. A bioreactor that provides mechanical stimuli was used to culture the magnetically seeded scaffolds over a 21-day period. Bioreactor maturation largely improved chondrogenesis into the cellularized scaffolds; the extracellular matrix obtained under these conditions was rich in collagen II and aggrecan. This work outlines the innovative potential of magnetic condensation of labeled stem cells and dynamic maturation in a bioreactor for improved chondrogenic differentiation, both scaffold-free and within polysaccharide scaffolds.

Chondrogenesis from stem cells requires fine tuning the culture conditions. Here, we present a magnetic approach for condensing cells, an essential step to initiate chondrogenesis. In addition, we show that dynamic maturation in a bioreactor applies mechanical stimulation to the cellular constructs and enhances cartilaginous extracellular matrix production.

3D agregación magnética Stem Cell y biorreactor de maduración para la regeneración del cartílago

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach ...

Bioingeniería

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result of chemical inputs to its synapses. However, neurons can also be excited by an imposed electric field. In particular, recent clinical applications activate neurons by creating an electric field externally. It is therefore of interest to investigate how the neuron responds to the external field and what causes the action potential. Fortunately, precise and controlled application of an external electric field is possible for embryonic neuronal cells that are excised, dissociated and grown in cultures. This allows the investigation of these questions in a highly reproducible system.
In this paper some of the techniques used for controlled application of external electric field on neuronal cultures are reviewed. The networks can be either one dimensional, i.e. patterned in linear forms or allowed to grow on the whole plane of the substrate, and thus two dimensional. Furthermore, the excitation can be created by the direct application of electric field via electrodes immersed in the fluid (bath electrodes) or by inducing the electric field using the remote creation of magnetic pulses.

Neuronal cultures are a good model for studying emerging brain stimulation techniques via their effect on single neurons or a population of neurons. Presented here are different methods for stimulation of patterned neuronal cultures by an electric field produced directly by bath electrodes or induced by a time-varying magnetic field.

La excitación externa de neuronas usando campos eléctricos y magnéticos en culturas de una y dos dimensiones

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result ...

Neurociencias

Electric and Magnetic Field Devices for Stimulation of Biological Tissues

Electric fields (EFs) and magnetic fields (MFs) have been widely used by tissue engineering to improve cell dynamics such as proliferation, migration, differentiation, morphology, and molecular synthesis. However, variables such stimuli strength and stimulation times need to be considered when stimulating either cells, tissues or scaffolds. Given that EFs and MFs vary according to cellular response, it remains unclear how to build devices that generate adequate biophysical stimuli to stimulate biological samples. In fact, there is a lack of evidence regarding the calculation and distribution when biophysical stimuli are applied. This protocol is focused on the design and manufacture of devices to generate EFs and MFs and implementation of a computational methodology to predict biophysical stimuli distribution inside and outside of biological samples. The EF device was composed of two parallel stainless-steel electrodes located at the top and bottom of biological cultures. Electrodes were connected to an oscillator to generate voltages (50, 100, 150 and 200 Vp-p) at 60 kHz. The MF device was composed of a coil, which was energized with a transformer to generate a current (1 A) and voltage (6 V) at 60 Hz. A polymethyl methacrylate support was built to locate the biological cultures in the middle of the coil. The computational simulation elucidated the homogeneous distribution of EFs and MFs inside and outside of biological tissues. This computational model is a promising tool that can modify parameters such as voltages, frequencies, tissue morphologies, well plate types, electrodes and coil size to estimate the EFs and MFs to achieve a cellular response.

This protocol describes the step-by-step process to build both electrical and magnetic stimulators used to stimulate biological tissues. The protocol includes a guideline to simulate computationally electric and magnetic fields and manufacture of stimulator devices.

Dispositivos eléctricos y de campo magnético para la estimulación de tejidos biológicos

Electric fields (EFs) and magnetic fields (MFs) have been widely used by tissue engineering to improve cell dynamics such as proliferation, migration, ...

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Agrobacterium-mediated transient protein production in plants is a promising approach to produce vaccine antigens and therapeutic proteins within a short period of time. However, this technology is only just beginning to be applied to large-scale production as many technological obstacles to scale up are now being overcome. Here, we demonstrate a simple and reproducible method for industrial-scale transient protein production based on vacuum infiltration of Nicotiana plants with Agrobacteria carrying launch vectors. Optimization of Agrobacterium cultivation in AB medium allows direct dilution of the bacterial culture in Milli-Q water, simplifying the infiltration process. Among three tested species of Nicotiana, N. excelsiana (N. benthamiana &#215; N. excelsior) was selected as the most promising host due to the ease of infiltration, high level of reporter protein production, and about two-fold higher biomass production under controlled environmental conditions. Induction of Agrobacterium harboring pBID4-GFP (Tobacco mosaic virus-based) using chemicals such as acetosyringone and monosaccharide had no effect on the protein production level. Infiltrating plant under 50 to 100 mbar for 30 or 60 sec resulted in about 95% infiltration of plant leaf tissues. Infiltration with Agrobacterium laboratory strain GV3101 showed the highest protein production compared to Agrobacteria laboratory strains LBA4404 and C58C1 and wild-type Agrobacteria strains at6, at10, at77 and A4. Co-expression of a viral RNA silencing suppressor, p23 or p19, in N. benthamiana resulted in earlier accumulation and increased production (15-25%) of target protein (influenza virus hemagglutinin).

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Transient protein production in Nicotiana plants based on vacuum infiltration with Agrobacteria carrying launch vectors (Tobacco mosaic virus-based) is a rapid and economic approach to produce vaccine antigens and therapeutic proteins. We simplified the procedure and improved target accumulation by optimizing conditions of bacteria cultivation, selecting host species, and co-introducing RNA silencing suppressors.

La optimización y utilización de<em&gt; Agrobacterium</em&gt; Mediada por la producción de proteína transitoria en<em&gt; Nicotiana</em

Agrobacterium-mediated transient protein production in plants is a promising approach to produce vaccine antigens and therapeutic proteins within a short ...

Biología

Growing Magnetotactic Bacteria of the Genus Magnetospirillum: Strains MSR-1, AMB-1 and MS-1

Magnetotactic bacteria are Gram-negative, motile, mainly aquatic prokaryotes ubiquitous in freshwater and marine habitats. They are characterized by their ability to biomineralize magnetosomes, which are magnetic nanometer-sized crystals of magnetite (Fe3O4) or greigite (Fe3S4) surrounded by a lipid bilayer membrane, within their cytoplasm. For most known magnetotactic bacteria, magnetosomes are assembled in chains inside the cytoplasm, thereby conferring a permanent magnetic dipole moment to the cells and causing them to align passively with external magnetic fields. Because of these specific features, magnetotactic bacteria have a great potential for commercial and medical applications. However, most species are microaerophilic and have specific O2 concentration requirements, making them more difficult to grow routinely than many other bacteria such as Escherichia coli. Here we present detailed protocols for growing three of the most widely studied strains of magnetotactic bacteria, all belonging to the genus Magnetospirillum. These methods allow for precise control of the O2 concentration made available to the bacteria, in order to ensure that they grow normally and synthesize magnetosomes. Growing magnetotactic bacteria for further studies using these procedures does not require the experimentalist to be an expert in microbiology. The general methods presented in this article may also be used to isolate and culture other magnetotactic bacteria, although it is likely that growth media chemical composition will need to be modified.

Growing Magnetotactic Bacteria of the Genus Magnetospirillum: Strains MSR-1, AMB-1 and MS-1

We present a procedure for growing several strains of Magnetospirillum in two different types of growth media. Magnetospirillum gryphiswaldense strain MSR-1 is grown in both liquid and O2 concentration gradient semi-solid media while M. magneticum strain AMB-1 and M. magnetotacticum strain MS-1 are grown in liquid medium.

Cultivo de bacterias magnétotactiques del género Magnetotacticum: cepas MSR-1, 1 AMB y MS-1

Magnetotactic bacteria are Gram-negative, motile, mainly aquatic prokaryotes ubiquitous in freshwater and marine habitats. They are characterized by their ...

MRM Microcoil Performance Calibration and Usage Demonstrated on Medicago truncatula Roots at 22 T

This protocol describes a signal-to-noise ratio (SNR) calibration and sample preparation method for solenoidal microcoils combined with biological samples, designed for high-resolution magnetic resonance imaging (MRI), also referred to as MR microscopy (MRM). It may be used at pre-clinical MRI spectrometers, demonstrated on Medicago truncatula root samples. Microcoils increase sensitivity by matching the size of the RF resonator to the size of the sample of interest, thereby enabling higher image resolutions in a given data acquisition time. Due to the relatively simple design, solenoidal microcoils are straightforward and cheap to construct and can be easily adapted to the sample requirements. Systematically, we explain how to calibrate new or home-built microcoils, using a reference solution. The calibration steps include: pulse power determination using a nutation curve; estimation of RF-field homogeneity; and calculating a volume-normalized signal-to-noise ratio (SNR) using standard pulse sequences. Important steps in sample preparation for small biological samples are discussed, as well as possible mitigating factors such as magnetic susceptibility differences. The applications of an optimized solenoid coil are demonstrated by high-resolution (13 x 13 x 13 &#956;m3, 2.2 pL) 3D imaging of a root sample.

MRM Microcoil Performance Calibration and Usage Demonstrated on Medicago truncatula Roots at 22 T

A protocol to study biological tissue at high spatial resolution using ultra-high field magnetic resonance microscopy (MRM) using microcoils is presented. Step-by-step instructions are provided for characterizing the microcoils. Finally, optimization of imaging is demonstrated on plant roots.

This protocol describes a signal-to-noise ratio (SNR) calibration and sample preparation method for solenoidal microcoils combined with biological ...

Stimulation of Patterned Neuronal Cultures by a Magnetic Field

Source: Stern, S., et al., <a href="https://app.jove.com/v/54357">External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures.</a> J. Vis. Exp. (2017)
This video demonstrates a method for stimulating neuronal cultures grown in a patterned circular ring on a coverslip, using a time-varying magnetic field.

Estimulación de cultivos neuronales modelados por un campo magnético

1. Preparation of coverslips for 1D cultures.

	Clean glass coverslips by immersion in a base piranha solution consisting of 75 mL DDW, 25 mL 25% ammonia ...

Mejora de la tasa de crecimiento inicial de las plantas agrícolas mediante el uso de campos magnéticos estáticos

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