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

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

1. Ajustes iniciais <ol><li> Espécies de plantas agrícolas <ol><li> Use Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) e Mescluns (Lepidium sativum) sementes. NOTA: Impatiens balsamina (Garden Balsam ou Rose Balsam) é uma espécie nativa para a Índia; alguns membros também estão localizados em Myanmar. Komatsuna (Brassica rapa var. Perviridis ou Komatsuna) é uma variante da mesma espécie que o nabo comum. Agrião de jardim (Lepidium sativum) é um tipo de erva que é taxonomicamente relacionadas com agrião e mostarda. Eles têm sabores semelhantes e cheiro, para que são comercialmente utilizados 5,7. </li></ol></li><li> As culturas de plantas <ol><li> Cultura Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) e Mescluns (Lepidium sativum) sementes em uma de 100 mm de diâmetro (100 pi) placa de Petri. Assegure-se que uma placa contém apenas um tipo de espécie. </li><li> Para condições de cultura, coloque as sementes em uma toalha de celulose. Imergir a toalha e sementes em água destilada tripla. Medir e confirmar que o interior laboratório RT é 18-25 ° C, com umidade variando de 65-75% (por favor, verifique o ponto 3.1.2). </li><li> Para o número de sementes, a cultura 10 ± 1 sementes de Garden Balsam, 50 ± 10 sementes de Mizuna, 330 ± 20 sementes de Komatsuna, e 380 ± 20 sementes de Mescluns. Use condições idênticas medidas como 18-25 ° C, com umidade variando de 65-75% (por favor, verifique o ponto 2.1.1). NOTA: Todos os experimentos foram realizados em condições interiores com a umidade regulada e faixa de temperatura em laboratório. A umidade ea temperatura não era estático, mas desde que as condições idênticas para o grupo e controle ímã tratada. </li></ol></li></ol> 2. Cultura de quatro plantas agrícolas<ol><li> Procedimento experimental <ol><li> Siga seção 1.2.3) para as espécies de condições da planta e da cultura em grupo controle e aplicado ímã. </li><li> Aplicar três imans de 1750 ± 350 Gauss (10.000 Gauss = 1 Tesla) na parte inferior das placas de 100 pi para Garden Balsam. Durante a aplicação, assegurar que as três ímans não estão em contacto directo com as sementes, e são separados por o fundo de plástico da placa de Petri. A distância direta entre sementes e ímãs deve ser 2-4 mm. Aplicar ímãs para 168 horas (7 dias) para quatro plantas agrícolas. </li><li> Seguindo todos os passos de forma idêntica em 2.1.2), aplicam-se dois magnetos, um (lado virado para cima N) no lado íman superior e outra (de frente para S para cima) na parte inferior da placa de cultura Garden Balsam. NOTA: Os pólos são aplicadas de forma diferente em Garden Balsam. No entanto, a orientação pólo não é considerado como um factor fundamental neste estudo para alterações do crescimento, como todos os ambientes são idênticos, excepto para a direcçãodo fluxo magnético. O objetivo do N e S aplicação pole para Garden Balsam foi ver a sua capacidade prática em usá-lo nos campos, onde a orientação pólo poderia ser difícil de gerir. </li></ol></li></ol> 3. Coloração de tubulina Jardim Balsam <ol><li> Ímã aplicações com condições de luz Regulado <ol><li> Coloque dois ímãs (N pólo para cima) inferior da placa de 100 mm para 48 horas, utilizando condições no passo 1.2.2. NOTA: Para a modificação de luz, as placas de cultura foram colocadas numa prateleira de plástico na incubadora. Incubadora foi usada para intercepção de luz e mantendo a temperatura a 25 ° C durante 48 h, em ambientes escuros. Eventualmente, esta condição não foi utilizado nesta experiência devido a variações elevadas de comprimento crescimento. </li></ol></li><li> coloração planta <ol><li> Fixar o impatiens inteiras SPP planta dupla flor, (incluindo caule e raízes) cultivadas em condições idênticas com o passo 3.1.2) em 4% de paraformaldeído e 0,1 Mtampão de fosfato (pH 7,4) durante 15 min. </li><li> Retirar a amostra e Impatiens imergir durante 2 h em tampão (2% de soro de cavalo / 1% de albumina de soro de bovino / 0,1% de Triton X-100 em PBS, pH 7,5) de bloqueio. Lava-se a amostra de Impatiens por imersão com PBS durante 15 min. </li><li> Para double imunocoloração, incubar a amostra com o anticorpo primário, anti-alfa tubulina (1: 1.000), O / N a 4 ° C. </li><li> Retirar a amostra e imergir as amostras uma vez com PBS durante 10 min para lavagem. Uso de FITC-conjugado anti-IgG de ratinho (1: 400) como anticorpo secundário e incubar durante 2 horas a 25 ° C. </li><li> Imergir a amostra em PBS e cobrir deslizar toda a amostra na parte inferior da placa de 24 poços. Obter imagens usando um microscópio de fluorescência convencional para observar a orientação tubulina (λ = 550 nm, ampliado a 100X, 200X e 400X). NOTA: Neste caso, o grupo ímã tratado (n = 10) e controles (n = 11) foram confirmados para Garden Balsam (Impatiens balsamina) somente, cultivadas em condições não-escuras. </li&gt; </ol></li></ol> 4. Métodos de Recolha de Dados <ol><li> Criação de lapso de tempo de Crescimento Quatro plantas agrícolas <ol><li> Fotografar a planta em intervalos de 10 min, através da criação do obturador para auto (isso pode ser feito em qualquer câmera digital). Defina a abertura para f 3.2 e o valor ISO para 400. </li><li> Recolha 700-900 fotos por 7-10 dias. Ligue a câmera com fios elétricos desde bateria pode estar esgotada. </li><li> Arraste as imagens clicando e soltando cada imagem cronologicamente abaixo da linha de streaming com software de criação de filmes (ver materiais e mesa de equipamento). Coloque-o em uma linha de streaming em durações iguais de ,045-,05 seg para cada em um total de filme de 30-40 seg. Verifique para que não haja lacunas escuras com selecionando a cada imagem em uma ordem cronológica. </li><li> Após o passo 4.1.3, clique no botão jogo no software para assegurar o filme compilados em um lapso de tempo de slides de vídeo 30-40 seg e clique renderizar e salvar em .mpeg ou em formato .avi. Para o tamanho markers, usar um quarto canadiana, um Penny americana, e uma régua centímetros no lado da foto. </li><li> Realizar teste t e gráfico de caixa para análise estatística 11,12. NOTA: Grupos de resumos de cinco números foram utilizados para calcular o limite inferior (L) valor como Q1 - [1,5 x (Q3 - Q1)] eo limite superior (H) valor como Q3 + [1,5 x (Q3 - Q1) ]. Esta abordagem foi incorporada passo 1.2.2 para a coleta de dados de comprimento 11. Os valores de L e H mostram a área de 99% dos lotes distribuição-t, o que significa valores aberrantes que os pontos dos dados observados do lado de fora desta gama podem ser considerados. Os diagramas de caixa e de Student t -test foram usados ​​para analisar as diferenças nas alturas das mudas de 12. </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.

Em todas as condições, ímãs devem ser aplicados sob a placa de Petri. Este estudo examinou a influência de campos magnéticos sobre a taxa de crescimento das sementes de várias espécies agrícolas, com foco no Garden Balsam como representante de plantas agrícolas. Por exemplo, a coloração tubulina foi realizada sobre Jardim de bálsamo para avaliar as alterações de nível molecular na raiz e caule micro-estruturas esqueléticas, sugerindo influência do campo magnético na proliferação de comprimento. Ambo...

A germinação é o crescimento de uma planta que resulta na formação de plântulas a 1. Sob certas condições, começa a germinação das sementes e nos tecidos embrionários retomar o crescimento. Ela começa com hidratação para a semente, a fim de activar enzimas para a germinação. As sementes podem ser induzidas a germinar in vitro (em uma placa de Petri ou tubo de ensaio) 1,2. Campos magnéticos estáticos são forças especiais que causam movimentos de moléculas com cargas iônicas por meio da força de Lorentz 3,4. força de Lorentz é formado quando um ionizados ou acusadas objeto se move sob um campo magnético. Todo o material é formado com os átomos que são compostos de electrões e protões. Quando os campos magnéticos tornam-se presentes, se é estático ou alternada, que afeta o movimento do material carregado. Isto também se aplica às plantas e moléculas de água, que afeta a condição molécula intracelular. Em um estudo anterior, foram utilizados bobinas eletromagnéticaspara gerar campos magnéticos pulsantes, e as plantas &#39;Komatsuna&#39; foram escolhidos como os indivíduos 5. No presente estudo, o ímã gerados campos magnéticos estáticos foram usadas para dar uma efeitos semelhantes, mas diferentes, como um estudo de expansão da força de Lorentz. A frequência do campo magnético, ao invés de sua polaridade, é um factor crucial para a germinação da planta. Estudos anteriores sugeriram que as taxas máximas de germinação foram 20% maior do que o controle quando a frequência do campo magnético foi de aproximadamente 10 Hz. Quando o campo foi removido de uma maneira retrógrada, a taxa de crescimento foi prejudicada 5. Campos magnéticos estáticos ter um efeito considerável sobre o crescimento inicial de 6-8, principalmente na germinação 6 e raiz de crescimento de 7. No presente estudo, foram utilizados magnetos estáticos para examinar a possibilidade de regular o crescimento de plantas agrícolas, utilizando campos magnéticos. Em particular, o nosso objectivo determine se determinadas condições de aplicação do campo magnético poderia aumentar as taxas de crescimento para níveis mais elevados do que os mencionados na literatura. Além disso, se a germinação inicial das plantas pode ser aumentada com sucesso utilizando um campo magnético, a utilização de fertilizantes químicos pode ser evitado.

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

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

dispositivos eletrônicos e fios de alta tensão induzem campos magnéticos. Um campo magnético de 1,300-2,500 Gauss (0,2 Tesla) foi aplicado a placas de Petri contendo sementes de Garden Balsam (Impatiens balsamina), Mizuna (Brassica rapa var. Japonica), Komatsuna (Brassica rapa var. Perviridis) e Mescluns (Lepidium sativum ). Nós aplicamos ímãs sob o prato de cultura. Durante os 4 dias de aplicação, observou-se que o comprimento do caule e raiz aumentou. O grupo submetido a tratamento de campo magnético (n = 10) mostrou um 1.4 vezes mais rápido taxa de crescimento em comparação com o grupo de controlo (n = 11) num total de 8 dias (p &lt;0,0005). Esta taxa é 20% maior do que a relatada em estudos anteriores. As linhas complexas de tubulina não têm pontos de conexão, mas os pontos de conexão ocorrer após a aplicação de ímãs. Esta diferença mostra completa a partir do controlo, o que significa arranjos anormais. No entanto, a causa exacta permanece por esclarecer. estes resÜLTS de intensificação do crescimento da aplicação imans sugerem que é possível aumentar a taxa de crescimento, aumentar a produtividade, ou controlar a velocidade de germinação de sementes de plantas através da aplicação de campos magnéticos estáticos. Também, os campos magnéticos pode causar alterações fisiológicas em células de plantas e podem induzir o crescimento. Portanto, a estimulação com um campo magnético pode ter efeitos possíveis que são semelhantes aos dos fertilizantes químicos, o que significa que o uso de fertilizantes pode ser evitado.

Coloração tubulina mostrou disperso ou diluído estruturas nas plantas cultivadas na presença do íman em comparação com o controlo (Figura 2). Além disso, 7 dias estudos de lapso de tempo com plantas agrícolas, incluindo Komatsuna (Brassica rapa var. Perviridis) e Mescluns (Lepidium sativum) indicou que um ímã derivado campo magnético estático aumenta o crescimento inicial das plantas (Figura 3). <p class="jove_content" ...

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.

Aperfeiçoamento da taxa de crescimento inicial das plantas agrícolas usando campos magnéticos estáticos

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

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13.2K visualizações.  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.

Vídeo: 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.

Geomagnética Field (Gmf) and Plant Evolution: investigação dos efeitos dos Gmf Reversão sobre<em&gt; Arabidopsis thaliana</em&gt; Desenvolvimento e Expressão Gênica

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

Biologia do Desenvolvimento

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.

Sensoriamento robótico e provisão de estímulos para crescimento de plantas guiadas

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

Engenharia

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 Agregação Stem Magnetic celular e biorreator Maturação para regeneração da cartilagem

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

Bioengenharia

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.

Excitação externa de Neurônios Usando Campos Elétricos e Magnéticos em One e culturas bidimensionais

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 ...

Neurociência

Electric and Magnetic Field Devices for Stimulation of Biological Tissues

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

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

Dispositivos de campo elétrico e magnético para estimulação de tecidos biológicos

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

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

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

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

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

Otimização e Utilização de<em&gt; Agrobacterium</em&gt; Mediada produção de proteína transiente em<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.

Crescimento de bactérias de Magnetotactic do género Magnetospirillum magnetotacticum: cepas 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.

Calibração e uso de desempenho da MRM Microcoil demonstrados em raízes de truncatula 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.

Estimulação de culturas neuronais padronizadas por um campo magnético

1. Preparation of coverslips for 1D cultures.

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

Aperfeiçoamento da taxa de crescimento inicial das plantas agrícolas usando campos magnéticos estáticos

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