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. Grundeinstellungen <ol><li> Landwirtschaftliche Pflanzenarten <ol><li> Verwenden Garten - Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) und Mescluns (Lepidium sativum) Samen. HINWEIS: Impatiens balsamina (Garten - Balsam oder Rose Balsam) ist eine Art stammt aus Indien; einige Mitglieder sind auch in Myanmar. Komatsuna (Brassica rapa var. Perviridis oder Komatsung) ist eine Variante der gleichen Art wie die gemeinsame Rübe. Gartenkresse (Lepidium sativum) ist eine Art von Kraut , das taxonomisch zu Kresse und Senf verwandt ist. Sie haben ähnliche Geschmacks- und Duft, für die sie 5,7 kommerziell genutzt werden. </li></ol></li><li> Pflanzenkulturen <ol><li> Kultur Garten - Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) und Mescluns (Lepidium sativum) Samen in einem Durchmesser von 100 mm (100 pi) Petrischale. Stellen Sie sicher, dass eine Platte nur eine Art von Spezies enthält. </li><li> Für Kulturbedingungen, legen Sie die Samen auf einem Zellulosetuch. Tauchen Sie das Handtuch und Samen in dreifach destilliertem Wasser. Messen Sie und bestätigen Sie, dass der Innen Labor RT ist 18-25 ° C, mit einer Luftfeuchtigkeit von 65-75% (bitte Abschnitt 3.1.2). </li><li> Für Anzahl von Samen, Kultur 10 ± 1 Samen von Garten-Balsam, 50 ± 10 Samen von Mizuna, 330 ± 20 Samen von Komatsuna und 380 ± 20 Samen von Mescluns. Verwenden Sie den gleichen Bedingungen gemessen, wie 18-25 ° C, mit einer Luftfeuchtigkeit von 65-75% (bitte Abschnitt 2.1.1). HINWEIS: Alle Experimente wurden an Bedingungen im Innenbereich mit der geregelten Luftfeuchtigkeit und Temperaturbereich im Labor durchgeführt. Die Feuchtigkeit und die Temperatur war nicht statisch, sondern vorgesehen, um die gleichen Bedingungen für den Magneten behandelten Gruppe und der Kontroll. </li></ol></li></ol> 2. Kultur von vier landwirtschaftlichen Pflanzen<ol><li> Versuchsdurchführung <ol><li> Folgen Sie Abschnitt 1.2.3) für Pflanzenarten und Kulturbedingungen in der Steuerung und Magnet angewendet Gruppe. </li><li> Bewerben drei Magnete von 1.750 ± 350 Gauss (10.000 Gauss = 1 Tesla) am unteren Rand der 100 pi Gerichte für Garten-Balsam. Während der Anwendung sicher, dass die drei Magnete nicht in direktem Kontakt mit den Samen, und werden durch den Kunststoffboden der Petrischale getrennt. Der direkte Abstand zwischen Samen und Magneten sollte 2-4 mm betragen. Tragen Sie Magnete für 168 Stunden (7 Tage) für vier landwirtschaftlichen Pflanzen. </li><li> Im Anschluss an alle identisch Schritte in 2.1.2), gelten zwei Magneten, ein (gegenüber N-Seite nach oben) auf der Oberseite und anderen Magneten (gegenüber S-Seite nach oben) an der Unterseite des Garten-Balsam Kulturplatte. HINWEIS: Die Pole sind unterschiedlich in Garten-Balsam aufgetragen. Jedoch ist die Polausrichtung nicht als entscheidender Faktor in dieser Studie für das Wachstum Veränderungen berücksichtigt, da alle Umgebungen identisch mit Ausnahme der Richtung sind,des magnetischen Flusses. Der Zweck von N- und S-Pol-Anwendung für Garten-Balsam war sich mit der es in den Bereichen der praktischen Fähigkeit, zu sehen, wo Pole Orientierung schwer sein könnte, zu verwalten. </li></ol></li></ol> 3. Tubulin Anfärbung von Garten-Balsam <ol><li> Magnet-Anwendungen mit geregelten Licht Zustand <ol><li> Legen Sie zwei Magnete (N-Pol nach oben) unten in der 100-mm-Platte für 48 Stunden unter Verwendung von Bedingungen in Schritt 1.2.2. HINWEIS: Für die Modifikation von Licht, die Kulturschalen auf einem Kunststoff-Regal in den Inkubator gelegt wurden. Inkubator wurde das Abfangen von Licht und Halten der Temperatur bei 25 ° C für 48 Stunden in dunklen Umgebungen verwendet wird. Schließlich wurde diese Bedingung nicht bei diesem Versuch durch hohe Schwankungen in Wachstumslänge. </li></ol></li><li> Pflanzen Anfärben <ol><li> Befestigen Sie das gesamte Impatiens SPP Doppel Blume Pflanze (einschließlich Stamm und Wurzeln) gewachsen auf identischen Bedingungen mit Schritt 3.1.2) in 4% Paraformaldehyd und 0,1 MPhosphatpuffer (pH 7,4) für 15 min. </li><li> Entfernen Sie die Impatiens Probe und tauche für 2 h in Blockierungspuffer (2% Pferdeserum / 1% Rinderserumalbumin / 0,1% Triton X-100 in PBS, pH 7,5). Waschen Sie die Impatiens Probe für 15 min mit PBS eingetaucht wird. </li><li> Für Doppel-Immunfärbung, Inkubation der Probe mit dem primären Antikörper, anti-alpha-Tubulin (1: 1000), O / N bei 4 ° C. </li><li> Entfernen Sie die Probe und tauchen Sie die Proben einmal mit PBS für 10 Minuten zu waschen. Verwendung FITC-konjugierter anti-Maus-IgG (1: 400) als sekundärer Antikörper und bei 25 ° C für 2 Stunden inkubiert. </li><li> Tauchen Sie die Probe in PBS und decken die gesamte Probe im Boden von 24-Well-Platte gleiten. Erhalten Sie Bilder ein herkömmliches Fluoreszenzmikroskop mit Tubulin Orientierung zu beobachten (λ = 550 nm, vergrößert zu 100X, 200X und 400X). HINWEIS: In diesem Fall wird der Magnet behandelten Gruppe (n = 10) und Kontrollgruppe (n = 11) nur für Garten - Balsam (Impatiens balsamina) bestätigt wurden, in nicht-Dunkel - Bedingungen gezüchtet. </li&gt; </ol></li></ol> 4. Methoden der Datenerhebung <ol><li> Zeitraffer Schaffung von vier landwirtschaftliche Pflanzenwachstum <ol><li> Fotografieren Sie das Werk in 10-Minuten-Intervallen, durch Verschluss Auto-Einstellung (dies kann in jeder digitalen Kamera gemacht werden). Stellen Sie die Blende F 3.2 und den ISO-Wert bis 400. </li><li> Sammeln Sie 700 bis 900 Bilder für 7-10 Tage. Schließen Sie die Kamera mit elektrischen Leitungen, da Batterie erschöpft sein könnte. </li><li> Ziehen Sie die Bilder durch einen Klick und Ablegen jedes Bild chronologisch unter der Streaming-Linie mit Filme Software (siehe Materialien und Geräte Tabelle). Legen Sie es auf einem Streaming-Linie in gleicher Dauer von 0,045 bis 0,05 sec für jeden in einen Film insgesamt 30-40 sec. Überprüfen Sie so, dass es keine dunklen Lücken mit dem jedes Bild in einer chronologischen Reihenfolge auswählen. </li><li> Nach dem Schritt 4.1.3, Play-Taste in der Software klicken Sie auf die kompilierte-Film in eine 30-40 sec Zeitraffer-Video-Folie und klicken Sie auf machen und speichern zu .mpeg oder .avi-Format zu gewährleisten. Für Größe markers Verwenden Canadian Quarter, einen amerikanischen Penny und einen Zentimeter Lineal auf der Seite des Fotos. </li><li> Führen t -test und Box - Plot für die statistische Analyse 11,12. HINWEIS: Gruppen von fünf Nummer Zusammenfassungen wurden verwendet, um die untere Grenze (L) Wert als die Berechnung von Q1 - [1,5 x (Q3 - Q1)] und die obere Grenze (H) Wert als Q3 + [1,5 x (Q3 - Q1) ]. Dieser Ansatz wurde in den Schritt integriert 1.2.2 für Länge der Datenerhebung 11. Die L- und H-Werte zeigen die 99% der Fläche der T-Verteilung Plots, was bedeutet, dass die Datenpunkte außerhalb dieses Bereichs beobachtet wird, kann als Ausreißer werden. Box - Plots und Student-t - Test verwendet wurden 12 , die Unterschiede in den Höhen von Sämlingen zu analysieren. </li></ol></li></ol>

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	<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.

In allen Bedingungen sollte Magneten unter der Petrischale aufgebracht werden. Diese Studie untersuchte den Einfluss von Magnetfeldern auf die Wachstumsrate der Samen für mehrere landwirtschaftliche Pflanzenarten, mit Fokus auf Garten-Balsam als Vertreter der landwirtschaftlichen Pflanzen. Zum Beispiel wurde Tubulin-Färbung an Garden Balsam durchgeführt, die molekularen Ebene Änderungen in root zu bewerten und Skelettmikrostrukturen stammen Einfluß des Magnetfeldes in der Länge Proliferation hindeutet. Sowohl die ...

Keimung ist das Wachstum einer Pflanze, die 1 in der Bildung der Keimling führt. Unter bestimmten Bedingungen beginnt die Keimung der Samen und die embryonalen Gewebe Wachstum wieder aufzunehmen. Es beginnt mit Feuchtigkeit auf das Saatgut, um Enzyme, die für die Keimung zu aktivieren. Samen können induziert werden , in vitro zu keimen (in einer Petrischale oder Teströhrchen) 1,2. Statische Magnetfelder sind spezielle Kräfte, die Bewegungen von Molekülen mit ionischen Ladungen durch die Lorentzkraft 3,4 verursachen. Lorentzkraft wird gebildet, wenn ein ionisiertes oder geladene Objekt bewegt sich unter einem Magnetfeld. Jedes Material wird mit Atomen gebildet, die aus Elektronen und Protonen bestehen. Wenn magnetische Felder vorhanden sein, ob sie statisch oder Wechsel ist, wirkt sich die Bewegung von geladenen Materials. Dies gilt auch für Pflanzen und Wassermoleküle, die die intrazelluläre Molekül Zustand wirkt. In einer früheren Studie wurden elektromagnetische Spulen verwendetzu erzeugen Pflanzen gepulste Magnetfelder und &#39;Komatsuna&#39; wurden 5 als die Themen gewählt. In der vorliegenden Studie erzeugten Magnet statische Magnetfelder wurden verwendet, um eine ähnliche, aber unterschiedliche Effekte als Expansions Studium der Lorentzkraft zu geben. Die Frequenz des Magnetfeldes, sondern als seine Polarität, ist ein entscheidender Faktor für die Pflanzen Keimung. Frühere Studien haben vorgeschlagen, dass maximale Keimungsraten 20% höher als die Kontrolle waren, wenn die Frequenz des Magnetfeldes ungefähr 10 Hz war. Wenn das Feld in einer retrograden Weise entfernt wurde, wurde die Wachstumsrate 5 beeinträchtigt. Statische Magnetfelder haben eine beträchtliche Auswirkung auf die anfängliche Wachstums 6-8, in erster Linie auf die Keimung und Wurzelwachstum 6 7. In der vorliegenden Untersuchung verwendeten wir statischen Magneten die Möglichkeit der Regulierung des Wachstums von landwirtschaftlichen Pflanzen zu untersuchen, durch magnetische Felder verwenden. Insbesondere wollten wir auf determine, ob bestimmte Bedingungen der Magnetfeldaufwandmengen, die Wachstums erhöhen könnte in der Literatur erwähnt diejenigen auf ein höheres Niveau als. Außerdem, wenn die anfängliche Keimung der Pflanzen kann erfolgreich unter Verwendung eines magnetischen Feldes erhöht werden, die Verwendung von chemischen Düngemitteln vermieden werden.

<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

Elektronische Geräte und Hochspannungsleitungen induzieren Magnetfelder. Ein Magnetfeld von 1,300-2,500 Gauss (0,2 Tesla) auf Petrischalen aufgetragen wurde Samen von Garten - Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) enthält, und Mescluns (Lepidium sativum ). Wir wandten Magneten unter der Kulturschale. Während der 4 Tage Anwendungs ​​beobachteten wir, dass der Stamm und Wurzellänge erhöht. Die Gruppe Magnetfeldbehandlung unterzogen (n = 10) zeigte eine 1,4-fache schneller Wachstumsrate im Vergleich mit der Kontrollgruppe (n = 11) in insgesamt 8 Tage (p &lt;0,0005). Diese Rate ist 20% höher als die in früheren Studien berichtet. Die Tubulin komplexen Linien hatte keine Punkte verbinden, aber Verbindungsstellen auftreten bei der Anwendung von Magneten. Dies zeigt vollständige Differenz von der Kontrolle, die abnormal Anordnungen bedeutet. Jedoch bleibt die genaue Ursache nicht klar. Diese results Wachstumssteigerung Magneten des Anwendens legen nahe, dass es möglich ist, die Wachstumsrate zu verbessern, die Produktivität zu erhöhen, oder um die Geschwindigkeit der Keimung der Pflanzen steuern durch statische Magnetfelder anwenden. Auch Magnetfelder können physiologische Veränderungen in Pflanzenzellen verursachen und das Wachstum induzieren kann. Daher Stimulation mit einem Magnetfeld können mögliche Effekte, die denen von chemischen Düngemitteln ähnlich sind, was bedeutet, dass die Verwendung von Düngemitteln vermieden werden kann.

Tubulin - Färbung zeigte in Pflanzen in Gegenwart des Magneten dispergiert oder verdünnt Strukturen gewachsen im Vergleich zur Kontrolle (Abbildung 2). Außerdem 7 Tage Zeitrafferstudien mit landwirtschaftlichen Pflanzen einschließlich Komatsuna (Brassica rapa var. Perviridis) und Mescluns (Lepidium sativum) zeigte an, daß ein Magnet statisches Magnetfeld abgeleitet erhöht das anfängliche Wachstum dieser Pflanzen (Abbildung 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.

Verbesserung des Anfangswachstumsrate von landwirtschaftlichen Pflanzen durch statische Magnetfelder

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 ...

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Magnetic Forces and Fields

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13.2K Aufrufe.  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.

Serie: 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.

Erdmagnetfeld (GMF) und Pflanze Evolution: Untersuchung der Auswirkungen von Gmf Reversal auf<em&gt; Arabidopsis thaliana</em&gt; Entwicklung und Genexpression

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

Forschung

Entwicklungsbiologie

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.

Robotic Sensing und Stimuli-Regelung für geführtes Pflanzenwachstum

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

Ingenieurwesen

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 ...

Umwelt

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 Magnetic Stem Cell Aggregation und Bioreactor Ausreifung für Knorpelregeneration

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

Biotechnik

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.

Externe Anregung von Neuronen mittels elektrische und magnetische Felder in One- und Zweidimensionale Kulturen

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 ...

Neurowissenschaften

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.

Elektrische und magnetische Feldgeräte zur Stimulation biologischer Gewebe

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.

Optimierung und Nutzung von<em&gt; Agrobacterium</em&gt; Transient-vermittelte Proteinproduktion in<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 ...

Biologie

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.

Wachsende Magnetotaktische Bakterien der Gattung Magnetospirillum: Stämme MSR-1, AMB-1 und 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.

MRM Microcoil Performance Calibration and Usage demonstriert auf Medicago-Truncatula-Wurzeln bei 22 T

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.

Stimulation von gemusterten neuronalen Kulturen durch ein Magnetfeld

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 ...

Verbesserung des Anfangswachstumsrate von landwirtschaftlichen Pflanzen durch statische Magnetfelder

Zusammenfassung

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