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. Impostazioni iniziali <ol><li> Specie di piante agricole <ol><li> Usa Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), e Mescluns (Lepidium sativum) semi. NOTA: Impatiens balsamina (Giardino balsamo o Rose Balsam) è una specie originaria dell&#39;India; alcuni membri si trovano anche in Myanmar. Komatsuna (Brassica rapa var. Perviridis o Komatsuna) è una variante della stessa specie, come la rapa comune. Crescione inglese (Lepidium sativum) è un tipo di erba che è tassonomicamente legato al crescione e senape. Hanno sapori simili e profumi, per le quali sono commercialmente utilizzati 5,7. </li></ol></li><li> Culture di piante <ol><li> Culture Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), e Mescluns (Lepidium sativum) semi in un diametro di 100 mm (100 PI) capsula di Petri. Assicurarsi che una piastra contiene un solo tipo di specie. </li><li> Per le condizioni di coltura, mettere i semi su un asciugamano di cellulosa. Immergere la spugna e semi in acqua distillata tripla. Misurare e confermare che il laboratorio interno RT è 18-25 ° C, con umidità che vanno 65-75% (si prega di consultare la sezione 3.1.2). </li><li> Per il numero di semi, la cultura di 10 ± 1 Sementi di Balsam, 50 ± 10 semi di Mizuna, 330 ± 20 semi di Komatsuna, e 380 ± 20 semi di Mescluns. Utilizzare condizioni identiche misurato come 18-25 ° C, con umidità che vanno 65-75% (si prega di consultare la sezione 2.1.1). NOTA: Tutti gli esperimenti sono stati condotti su condizioni interne con l&#39;umidità regolamentato e range di temperatura in laboratorio. L&#39;umidità e temperatura non era statica, ma purché le condizioni identiche per magnete gruppo trattato e di controllo. </li></ol></li></ol> 2. Cultura di quattro impianti agricoli<ol><li> Procedura sperimentale <ol><li> Seguire la sezione 1.2.3) per le specie di condizioni dell&#39;impianto e della cultura nel controllo e magnete gruppo applicata. </li><li> Applicare tre magneti di 1,750 ± 350 Gauss (10.000 Gauss = 1 Tesla) nella parte inferiore dei 100 piatti pi per Garden Balsam. Durante l&#39;applicazione, assicurano che i tre magneti non sono in contatto diretto con i semi, e sono separati dal fondo di plastica della capsula di Petri. La distanza tra i semi diretta e magneti dovrebbe essere di 2-4 mm. Applicare magneti per 168 ore (7 giorni) per quattro piante agricole. </li><li> Seguendo tutti i passi identicamente a 2.1.2), applicare due magneti, uno (rivolta verso l&#39;alto lato N) sul lato superiore e altro magnete (rivolta verso l&#39;alto S) sul fondo della piastra di coltura Garden Balsam. NOTA: I pali sono applicate in modo diverso a Garden Balsam. Tuttavia, l&#39;orientamento polo non è considerato un fattore cruciale in questo studio per alterazioni di crescita, come tutti gli ambienti sono identiche tranne per la direzionedel flusso magnetico. Lo scopo di N e S applicazione poli per Garden Balsam è stato quello di vedere la sua capacità pratica nel suo utilizzo nei campi, in cui l&#39;orientamento polo potrebbe essere difficile da gestire. </li></ol></li></ol> 3. tubulina colorazione di Garden Balsam <ol><li> Le applicazioni magnetiche con regolamentato Condizione Luce <ol><li> Inserire due magneti (N polo positivo in alto) inferiore della piastra di 100 mm per 48 ore, utilizzando le condizioni al punto 1.2.2. NOTA: Per modifica di luce, i piatti di coltura sono stati posti su un ripiano di plastica in incubatrice. Incubatore è stato utilizzato per l&#39;intercettazione della luce e temperatura mantenendo a 25 ° C per 48 ore in ambienti scuri. Infine, tale condizione non è stato utilizzato in questo esperimento a causa di forti variazioni di lunghezza crescita. </li></ol></li><li> La colorazione delle piante <ol><li> Fissare l&#39;intero Impatiens SPP impianto fiore doppio, (tra cui stelo e radici) coltivate su identiche condizioni con il punto 3.1.2) in paraformaldeide al 4% e 0,1 Mtampone fosfato (pH 7,4) per 15 min. </li><li> Rimuovere il campione Impatiens ed immergere per 2 ore in tampone di bloccaggio (2% siero di cavallo / 1% di siero albumina bovina / 0,1% Triton X-100 in PBS, pH 7,5). Lavare il campione impatiens immergendo con PBS per 15 min. </li><li> Per il doppio immunocolorazione, incubare il campione con l&#39;anticorpo primario, anti-alfa tubulina (1: 1.000), O / N a 4 ° C. </li><li> Rimuovere il campione ed immergere i campioni una volta con PBS per 10 minuti per lavare. Utilizzare FITC-coniugato anti-IgG di topo (1: 400) come anticorpo secondario e incubare per 2 ore a 25 ° C. </li><li> Immergere il campione in PBS e coprire scivolare tutto il campione in fondo a 24 pozzetti. Ottenere immagini utilizzando un microscopio a fluorescenza convenzionali per osservare l&#39;orientamento tubulina (λ = 550 nm, ingrandita a 100X, 200X e 400X). NOTA: In questo caso, il gruppo magnete trattati (n = 10) e controlli (n = 11) sono stati confermati per Garden Balsam (Impatiens balsamina) soltanto, coltivate in condizioni non-scuri. </li&gt; </ol></li></ol> Metodi 4. Data Collection <ol><li> Creazione Time-lapse della crescita Quattro piante agricole <ol><li> Fotografare l&#39;impianto ad intervalli di 10 min, impostando otturatore per auto (questo può essere fatto in qualsiasi fotocamera digitale). Impostare l&#39;apertura a F 3.2 e il valore ISO 400. </li><li> Raccogliere 700-900 immagini per 7-10 giorni. Collegare la fotocamera con i fili elettrici da batteria potrebbe essere esaurita. </li><li> Trascinare le immagini facendo clic e rilasciando ogni foto in ordine cronologico sotto la linea di streaming con il software moviemaking (vedi Materiali e attrezzature tabella). Mettilo su una linea di flusso in uguali durate di 0,045-0,05 sec per ciascuna in un totale pellicola di 30-40 sec. Verificare in modo che non ci siano spazi scuri con la selezione del ogni immagine in un ordine cronologico. </li><li> Dopo il punto 4.1.3, fare clic sul pulsante di riproduzione nel software per assicurare la-movie compilato in un 30-40 secondi time-lapse video di presentazione e fare clic su di rendering e salvare .mpeg o in formato .avi. Per dimensioni markers, utilizzare Canadian trimestre, un Penny americano, e un righello centimetri sul lato della foto. </li><li> Eseguire t-test e la trama di dialogo per l&#39;analisi statistica 11,12. NOTA: Gruppi di sintesi di cinque numeri sono stati usati per calcolare il limite inferiore (L) valore Q1 - [1.5 × (Q3 - Q1)] e il più alto limite di (H) valore Q3 + [1,5 × (Q3 - Q1) ]. Questo approccio è stato incorporato nel passo 1.2.2 per la raccolta dati di lunghezza 11. I valori di L e H mostrano la zona il 99% degli appezzamenti T-distribuzione, il che significa valori anomali che i punti dati osservati al di fuori di questo intervallo può essere considerato. Trame Box e t di Student-test sono stati utilizzati per analizzare le differenze nelle altezze di piantine 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.

In tutte le condizioni, i magneti devono essere applicati sotto la capsula di Petri. Questo studio ha esaminato l&#39;influenza dei campi magnetici sul tasso di crescita di semi per diverse specie agricole, con particolare attenzione alla Garden Balsam come rappresentante di piante agricole. Per esempio, tubulina colorazione è stata eseguita su Garden Balsam valutare i cambiamenti a livello molecolare in radice e gambo micro-strutture scheletriche suggeriscono influenza del campo magnetico nella proliferazione lunghezz...

La germinazione è la crescita di una pianta che provoca la formazione della piantina 1. In determinate condizioni, la germinazione del seme inizia e tessuti embrionali riprendere la crescita. Si inizia con idratazione al seme per attivare gli enzimi per la germinazione. Semi possono essere indotte a germinare in vitro (in una capsula di Petri o provetta) 1,2. I campi magnetici statici sono forze speciali che causano movimenti di molecole con cariche ioniche a titolo della forza di Lorentz 3,4. forza di Lorentz si forma quando un oggetto ionizzati o cariche si muove sotto un campo magnetico. Ogni materiale è formato con atomi che sono composti da elettroni e protoni. Quando campi magnetici diventano presenti, se è statico o alternata, colpisce il movimento del materiale caricato. Questo vale anche per piante e molecole di acqua, che colpisce la condizione molecola intracellulare. In uno studio precedente, sono stati utilizzati bobine elettromagneticheper generare campi magnetici pulsati, e le piante &#39;Komatsuna&#39; sono stati scelti come i soggetti 5. Nel presente studio, magnete generato campi magnetici statici sono stati usati per dare un effetti simili ma diversi da uno studio espansione della forza di Lorentz. La frequenza del campo magnetico, piuttosto che la sua polarità, è un fattore cruciale per la germinazione delle piante. Precedenti studi hanno suggerito che i tassi massimi germinazione erano superiori del 20% rispetto al controllo quando la frequenza del campo magnetico era di circa 10 Hz. Quando il campo è stato rimosso in maniera retrograda, il tasso di crescita è stata compromessa 5. I campi magnetici statici hanno un effetto considerevole sul 6-8 crescita iniziale, in primo luogo sulla germinazione 6 e la radice della crescita 7. Nel presente studio, abbiamo utilizzato magneti statici per esaminare la possibilità di regolare la crescita delle piante agricole utilizzando i campi magnetici. In particolare, abbiamo voluto determine se certe condizioni di applicazione del campo magnetico potrebbe aumentare i tassi di crescita a livelli più elevati rispetto a quelli citati in letteratura. Inoltre, se la germinazione iniziale delle piante può essere aumentata con successo utilizzando un campo magnetico, l&#39;uso di fertilizzanti chimici può essere evitato.

<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

Dispositivi elettronici e cavi ad alta tensione inducono campi magnetici. Un campo magnetico di 1,300-2,500 Gauss (0,2 Tesla) è stata applicata a piastre di Petri contenenti semi di Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis), e Mescluns (Lepidium sativum ). Abbiamo applicato magneti sotto il piatto di cultura. Durante i 4 giorni di applicazione, abbiamo osservato che il fusto e della radice la lunghezza aumentata. Il gruppo sottoposto a trattamento campo magnetico (n = 10) ha mostrato un 1,4 volte più veloce tasso di crescita rispetto al gruppo di controllo (n = 11) in un totale di 8 giorni (p &lt;0,0005). Questo tasso è superiore del 20% rispetto a quello riportato in studi precedenti. Le linee complesse tubulina non hanno punti di collegamento, ma punti di collegamento si verificano al momento della domanda di magneti. Questo dimostra completa differenza dal controllo, che significa arrangiamenti anormali. Tuttavia, la causa esatta rimane poco chiaro. questi resULT di valorizzazione crescita di applicare magneti suggeriscono che è possibile migliorare il tasso di crescita, aumentare la produttività, o controllare la velocità di germinazione di piante applicando campi magnetici statici. Inoltre, i campi magnetici possono causare cambiamenti fisiologici nelle cellule vegetali e possono indurre la crescita. Pertanto, la stimolazione con un campo magnetico può avere effetti che sono simili a quelli di concimi chimici, il che significa che l&#39;uso di fertilizzanti può essere evitato.

Tubulina colorazione mostrato disperso o strutture diluito in piante coltivate in presenza del magnete rispetto al controllo (Figura 2). Inoltre, gli studi time-lapse 7 giorni con piante agricole tra cui Komatsuna (Brassica rapa var. Perviridis) e Mescluns (Lepidium sativum) hanno indicato che un magnete derivato campo magnetico statico aumenta la crescita iniziale di queste piante (Figura 3). <p class="jove_content" fo:keep-togethe...

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.

Valorizzazione del tasso di crescita iniziale delle Piante Agrarie mediante l&#39;utilizzo di campi magnetici statici

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

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Magnetic Forces and Fields

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13.2K Views.  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.

Geomagnetico Field (Gmf) e Stabilimento Evolution: studio degli effetti di Gmf Storno su<em&gt; Arabidopsis thaliana</em&gt; Sviluppo e Gene Expression

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

Ricerca

Biologia dello sviluppo

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.

Rilevamento robotico e fornitura di stimoli per la crescita guidata delle piante

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

Ingegneria

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 ...

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 Stem magnetica cellulare Aggregazione e bioreattore maturazione per la cartilagine Regeneration

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

Bioingegneria

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.

Eccitazione esterna dei neuroni Usando campi elettrici e magnetici in mono e culture bidimensionali

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 ...

Neuroscienze

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.

Dispositivi elettrici e a campo magnetico per la stimolazione dei tessuti biologici

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.

Ottimizzazione e utilizzo di<em&gt; Agrobacterium</em&gt;-Mediata transitoria produzione di proteine ​​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 ...

Biologia

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.

Crescita di batteri magnetotattici del genere Magnetospirillum: ceppi MSR-1, AMB-1 e 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.

Calibrazione e utilizzo delle prestazioni del microcoil MRM dimostrato sulle radici della trogola Medicago a 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.

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 ...

Valorizzazione del tasso di crescita iniziale delle Piante Agrarie mediante l'utilizzo di campi magnetici statici

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