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. Réglages initiaux <ol><li> Espèces de plantes agricoles <ol><li> Utiliser Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), et Mescluns (Lepidium sativum). NOTE: Impatiens balsamina (jardin Balsam ou Rose Balsam) est une espèce indigène à l&#39; Inde; quelques membres sont également situés au Myanmar. Komatsuna (Brassica rapa var. Perviridis ou komatsuna) est une variante de la même espèce que le navet commun. Cress Garden (Lepidium sativum) est un type de plante qui est taxonomiquement liée à cresson et la moutarde. Ils ont des saveurs et des parfums similaires, pour lesquels ils sont utilisés dans le commerce de 5,7. </li></ol></li><li> Cultures végétales <ol><li> Culture Jardin Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), et Mescluns (Lepidium sativum) graines dans un diamètre de 100 mm (100 pi) boîte de Pétri. Assurez-vous que une plaque ne contient qu&#39;un seul type d&#39;espèce. </li><li> Pour des conditions de culture, placer les graines sur une serviette en cellulose. Immergez la serviette et les graines dans de l&#39;eau distillée triple. Mesurer et confirmer que le laboratoire intérieur RT est 18-25 ° C, avec une humidité allant de 65-75% (s&#39;il vous plaît consulter la section 3.1.2). </li><li> Pour nombre de graines, de la culture 10 ± 1 graines de jardin Balsam, 50 ± 10 graines de Mizuna, 330 ± 20 graines de Komatsuna, et 380 ± 20 graines de Mescluns. Utiliser des conditions identiques mesurées comme 18-25 ° C, avec une humidité allant de 65-75% (s&#39;il vous plaît consulter la section 2.1.1). NOTE: Toutes les expériences ont été réalisées sur des conditions intérieures avec l&#39;humidité régulée et plage de température dans le laboratoire. L&#39;humidité et la température était pas statique, mais à condition que les conditions identiques pour aimant traité et le groupe témoin. </li></ol></li></ol> 2. Culture des Quatre plantes agricoles<ol><li> Procédure expérimentale <ol><li> Suivez la section 1.2.3) pour les espèces de conditions de plantes et de la culture dans le contrôle et le groupe appliqué à aimant. </li><li> Appliquer trois aimants de 1.750 ± 350 Gauss (10.000 Gauss = 1 Tesla) au fond des boîtes de 100 pi pour Garden Balsam. Lors de l&#39;application, en sorte que les trois aimants ne sont pas en contact direct avec les semences, et sont séparés par le fond en plastique de la boîte de Pétri. La distance directe entre les semences et les aimants doit être de 2-4 mm. Appliquer des aimants pour 168 heures (7 jours) pour quatre plantes agricoles. </li><li> Après toutes les étapes identiques à 2.1.2), appliquer deux aimants, un (N côté tourné vers le haut) sur le côté de S supérieur et autre aimant (face vers le haut) sur le fond du jardin Balsam plaque de culture. NOTE: Les pôles sont appliqués différemment dans le jardin Balsam. Cependant, l&#39;orientation polaire est pas considéré comme un facteur crucial dans cette étude pour des altérations de la croissance, comme tous les environnements sont identiques, sauf pour la directiondu flux magnétique. Le but de N et S application pole pour Garden Balsam était de voir sa capacité pratique à l&#39;utiliser dans les champs, où l&#39;orientation pôle pourrait être difficile à gérer. </li></ol></li></ol> 3. tubuline Coloration de jardin Balsam <ol><li> Magnet Applications avec Condition Lumière Réglementé <ol><li> Placez deux aimants (N pôles orientés vers le haut) en bas de la plaque de 100 mm pendant 48 heures, à l&#39;aide de conditions à l&#39;étape 1.2.2. NOTE: Pour la modification de la lumière, les boîtes de culture ont été placés sur une étagère en plastique dans l&#39;incubateur. Incubateur a été utilisé pour l&#39;interception de la lumière et de maintien en température à 25 ° C pendant 48 heures dans des environnements sombres. Finalement, cette condition n&#39;a pas été utilisé dans cette expérience en raison de fortes variations de longueur de la croissance. </li></ol></li><li> Coloration des plantes <ol><li> Fixer le impatiens entiers SPP fleur double plante, (y compris les tiges et les racines) cultivées dans des conditions identiques à l&#39;étape 3.1.2) dans 4% de paraformaldehyde et 0,1 Mun tampon phosphate (pH 7,4) pendant 15 min. </li><li> Retirer l&#39;échantillon impatiens et immerge pendant 2 heures dans un tampon de blocage (2% de sérum de cheval / 1% de sérumalbumine bovine / 0,1% de Triton X-100 dans du PBS, pH 7,5). Laver l&#39;échantillon impatiens par immersion avec du PBS pendant 15 min. </li><li> Pour une double immunocoloration, incuber l&#39;échantillon avec un anticorps primaire, anti-alpha tubuline (1: 1000), O / N à 4 ° C. </li><li> Retirer l&#39;échantillon et immerger les échantillons une fois avec du PBS pendant 10 min pour se laver. L&#39;utilisation conjuguée à FITC anti-souris IgG (1: 400) comme anticorps secondaire et on incube pendant 2 heures à 25 ° C. </li><li> Immerger l&#39;échantillon dans du PBS et couvrir glisser l&#39;ensemble de l&#39;échantillon dans le fond de plaque à 24 puits. Obtenir des images en utilisant un microscope à fluorescence conventionnelle pour observer l&#39;orientation de la tubuline (λ = 550 nm, grossie à 100X, 200X et 400X). NOTE: Dans ce cas, le groupe d&#39;aimants traités (n = 10) et les contrôles (n = 11) ont été confirmés pour Garden Balsam (Impatiens balsamina) seulement, cultivé dans des conditions non sombres. </li&gt; </ol></li></ol> Méthodes 4. Collecte de données <ol><li> Time-lapse Création de croissance Quatre plantes agricoles <ol><li> Photographie de l&#39;usine à intervalles de 10 min, en réglant l&#39;obturateur pour auto (cela peut être fait dans un appareil photo numérique). Réglez l&#39;ouverture à F 3.2 et la valeur ISO 400. </li><li> Collecter 700-900 images pendant 7-10 jours. Branchez l&#39;appareil photo avec des fils électriques depuis la batterie peut être déchargée. </li><li> Faites glisser les images en cliquant et en laissant tomber chaque image chronologiquement sous la ligne de streaming avec le logiciel moviemaking (voir matériaux et table d&#39;équipement). Mettez-le sur une ligne de streaming en durées égales de 0,045 à 0,05 s pour chaque en un total de film de 30-40 sec. Vérifiez afin qu&#39;il n&#39;y ait pas de lacunes sombres avec la sélection de la chaque image dans un ordre chronologique. </li><li> Après l&#39;étape 4.1.3, cliquez sur le bouton de lecture dans le logiciel pour assurer le-film compilé dans un time-lapse vidéo slide 30-40 sec et cliquez rendre et enregistrer sur .mpeg ou au format .avi. Pour la taille markers, utilisez Quarter canadien, un Penny américain, et une règle de centimètres sur le côté de la photo. </li><li> Effectuer t -test et boîte à moustaches pour l&#39; analyse statistique 11,12. NOTE: Les groupes de cinq nombre résumés ont été utilisés pour calculer la limite inférieure (L) valeur Q1 - [1,5 × (Q3 - Q1)] et plus de limite (H) valeur Q3 + [1,5 × (Q3 - Q1) ]. Cette approche a été incorporé dans l&#39; étape 1.2.2 pour la collecte de données de longueur 11. Les L et H montrent des valeurs de la zone de 99% des parcelles T de distribution, ce qui signifie que les points de données observées en dehors de cette gamme peuvent être considérés comme des valeurs aberrantes. Parcelles de boîte et de Student t -test ont été utilisées pour analyser les différences dans les hauteurs de semis 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.

Dans toutes les conditions, les aimants doivent être appliqués dans la boîte de Pétri. Cette étude a examiné l&#39;influence des champs magnétiques sur le taux de graines de croissance pour plusieurs espèces agricoles, en mettant l&#39;accent sur le jardin Balsam en tant que représentant des plantes agricoles. Par exemple, la tubuline coloration a été réalisée sur Garden Balsam pour évaluer les changements au niveau moléculaire dans la racine et la tige micro-structures squelettiques suggérant l&#39;infl...

La germination est la croissance d&#39;une plante qui aboutit à la formation de la plantule 1. Sous certaines conditions, la germination des graines commence et les tissus embryonnaires reprendre la croissance. Elle commence par l&#39;hydratation de la semence afin d&#39;activer les enzymes pour la germination. Les graines peuvent être amenées à germer in vitro (dans une boîte de Pétri ou tube à essai) 1,2. Les champs magnétiques statiques sont des forces spéciales qui provoquent des mouvements de molécules avec des charges ioniques par l&#39; intermédiaire de la force de Lorentz 3,4. force de Lorentz est formé quand un ionisées ou chargées objet se déplace sous un champ magnétique. Chaque matériau est formé avec des atomes qui sont constituées d&#39;électrons et de protons. Lorsque les champs magnétiques deviennent présents, que ce soit statique ou alternatif, il affecte le mouvement du matériau chargé. Cela vaut également pour les plantes et les molécules d&#39;eau, ce qui affecte la condition molécule intracellulaire. Dans une précédente étude, bobines électromagnétiques ont été utiliséspour générer des champs magnétiques pulsés, et les plantes &#39;Komatsuna&#39; ont été choisis comme les sujets 5. Dans la présente étude, aimant généré des champs magnétiques statiques ont été utilisés pour donner des effets similaires mais différents comme une étude d&#39;expansion de la force de Lorentz. La fréquence du champ magnétique, plutôt que sa polarité, est un facteur crucial pour la germination des plantes. Des études antérieures ont suggéré que les taux de germination maximales étaient de 20% plus élevé que le contrôle lorsque la fréquence du champ magnétique était d&#39;environ 10 Hz. Lorsque le champ est éliminé d&#39; une manière rétrograde, le taux de croissance a été altérée 5. Les champs magnétiques statiques ont un effet considérable sur le 6-8 de croissance initiale, principalement sur ​​la germination 6 et la croissance des racines 7. Dans la présente étude, nous avons utilisé des aimants statiques pour examiner la possibilité de réguler la croissance des plantes agricoles en utilisant des champs magnétiques. En particulier, nous avons cherché à déterminer si certaines conditions d&#39;application de champ magnétique pourrait augmenter les taux de croissance à des niveaux plus élevés que ceux qui sont mentionnés dans la littérature. En outre, si la germination initiale des plantes peut être augmentée avec succès en utilisant un champ magnétique, l&#39;utilisation d&#39;engrais chimiques peut être évitée.

<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

Les appareils électroniques et les fils à haute tension induisent des champs magnétiques. Un champ magnétique de 1,300-2,500 Gauss (0,2 Tesla) a été appliqué à des boîtes de Petri contenant des graines de jardin Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), et Mescluns (Lepidium sativum ). Nous aimants appliqué dans la boîte de culture. Pendant les 4 jours d&#39;application, nous avons observé que la tige et la racine de longueur augmentée. Le groupe soumis à un traitement de champ magnétique (n = 10) a montré une 1,4 fois plus rapide taux de croissance par rapport au groupe témoin (n = 11) dans un total de 8 jours (p &lt;0,0005). Ce taux est de 20% plus élevé que celui rapporté dans les études précédentes. Les lignes complexes tubuline n&#39;ont des points de connexion, mais les points de connexion se produisent lors de l&#39;application d&#39;aimants. Cela montre la différence complète de la commande, ce qui signifie des dispositions anormales. Cependant, la cause exacte reste incertaine. Ces resULTS d&#39;amélioration de l&#39;application d&#39;aimants de croissance suggèrent qu&#39;il est possible d&#39;augmenter le taux de croissance, d&#39;augmenter la productivité, ou de contrôler la vitesse de germination des plantes en appliquant des champs magnétiques statiques. En outre, les champs magnétiques peuvent provoquer des changements physiologiques dans les cellules végétales et peuvent induire la croissance. Par conséquent, une stimulation par un champ magnétique, peut avoir des effets possibles qui sont semblables à celles des engrais chimiques, ce qui signifie que l&#39;utilisation d&#39;engrais peut être évitée.

Tubuline coloration a montré une dispersion ou des structures aminci dans des plantes cultivées en présence de l&#39;aimant par rapport au témoin (figure 2). En outre, 7 jour des études time-lapse avec des plantes agricoles , y compris Komatsuna (Brassica rapa var. Perviridis) et Mescluns (Lepidium sativum) ont indiqué que un aimant dérivé champ magnétique statique augmente la croissance initiale de ces plantes (Figure 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.

Amélioration du taux de croissance initiale des plantes agricoles à l&#39;aide de champs magnétiques statiques

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

Universidad de Cartagena UDEC

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Unit 1

Magnetic Forces and Fields

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13.1K vues.  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.

Vidéo: 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.

Champ géomagnétique (FMV) et Evolution des végétaux: Étude des effets de la GMF Reprise sur<em&gt; Arabidopsis thaliana</em&gt; Développement et Gene Expression

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

Recherche

Biologie du développement

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.

Fourniture de détection robotique et de stimuli pour la croissance guidée des plantes

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

Ingénierie

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 ...

Environnement

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.

L&#39;agrégation magnétique de cellules souches 3D et bioréacteurs Maturation pour Cartilage régénération

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

Bio-ingénierie

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 Excitation Neurones Utilisation de champs électriques et magnétiques et des cultures dans One- à deux dimensions

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 ...

Neurosciences

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.

Dispositifs électriques et magnétiques de champ pour la stimulation des tissus biologiques

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.

Optimisation et l&#39;utilisation des<em&gt; Agrobacterium</em&gt; Médiée par la production de protéines transitoire dans<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.

Croissance des bactéries magnétotactiques du genre Magnetospirillum: souches MSR-1, 1-AMB et 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.

Étalonnage et utilisation des performances de microcoil MRM démontrés sur les racines de la truncatule Medicago à 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://appjove.unicartagenaproxy.elogim.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 de cultures neuronales structurées par un champ magnétique

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 ...

Amélioration du taux de croissance initiale des plantes agricoles à l'aide de champs magnétiques statiques

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