Data reported in this work are part of the USDA-ARS National Program 306 Project 'Values-Added Products from Cottonseed' research supported by the Agency's in-house funding. Publication of this paper is supported in part by the Journal of Visualized Experiments. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer. We acknowledge the constructive comments from JoVE science editor and peer reviewers in the review and revision process.

1. La semilla de algodón y productos a base de harina de soja (Figura 1) <ol><li> Obtener las materias primas, de semilla de algodón desgrasado y comidas de soja, de fuentes disponibles en el mercado. </li><li> Obtener la comida de trabajo mediante molienda de la harina desgrasada sólido en un molino de muestras ciclón para pasar un tamiz de acero de 0,5 mm 16. </li><li> Preparar comidas lavados de agua de las comidas de trabajo después de la extracción de agua (25 g de harina: 200 ml de agua) para separar los componentes solubles en agua en las comidas 21. </li><li> Preparar aislados de proteínas de las comidas de trabajo por extracción alcalina y precipitación ácida 16. </li></ol> 2. Preparación de las tiras de chapa de madera <ol><li> Chapas de madera cortada (1,59 mm de espesor) obtenidos de una fuente disponible comercialmente en tiras 25,4 mm de ancho por 88,9 mm de largo. </li><li> Lápiz marque una línea a través de la veta de la madera en el 25,4 mm (1.0 &quot;) de longitud de un extremo de cada tira Etiqueta de estas tiras adecuadamente con pruebas tratamientos o números 5.. -10 pares de madera se preparan para cada variable de la prueba. </li></ol> 3. Preparación de adhesivo suspensiones <ol><li> Calcular la cantidad de agua se lavó comida necesaria por las muestras de madera para las pruebas, por la tasa de aplicación (por ejemplo, 4 mg seco contenido cm -2) x área total de unión (por ejemplo, 581 cm 2 de 90 tiras de madera con área de unión 2,54 x 2,54 cm cada uno) más un 30% adicional para enoughness (es decir, 4 x 581 x 130% 3 g de agua se lavó comida para el ejemplo). </li><li> Mezcle el agua comida se lavó con agua desionizada (3:25 w / w), y revuelva con una barra de agitación magnética durante 2 horas en un vaso sellado con Parafilm. </li></ol> 4. Preparación de servidumbre de madera Muestras <ol><li> Cepille lechada adhesiva en un extremo de tiras de chapa de madera que cubren 2 25.4 mm (1.0 &quot;) de longitud Aire seco durante 10 - 15 min. O hasta de mal gusto. </li><li> Cepillo una segunda capa de lechada de adhesivo en la parte superior de la primera capa y secar al aire de nuevo. La cantidad de adhesivo secopreparación aplicada es de alrededor de 4,5 mg de sólido seco por cm2 de unión son de cada tira de madera. </li><li> La superposición de la zona recubierta adhesivo pegajoso (25,4 x 25,4 mm o 1,0 &quot;x 1,0&quot;) de tiras de chapa de madera 2. Prensado en caliente utilizando una prensa de sobremesa climatizada a 100 ° C durante 20 min a una presión de 400 psi (2,8 MPa). Tenga en cuenta que la presión es la fuerza aplicada por la prensa dividida por el área superpuesta de las muestras de madera. Estos parámetros de unión pueden ser modificados según sea necesario para cada variable de la prueba. </li><li> Dejar enfriar y acondicionar las muestras de madera unidas durante 48 horas en un cuarto acondicionado o una incubadora con control de humedad (temperatura de 22-23 ° C y una humedad relativa del 50 - 60%; Figura 2). </li></ol> 5. Los experimentos resistencia al agua <ol><li> Sumergir las muestras de madera unidas, después del acondicionamiento inicial, en el agua del grifo durante 48 horas en una bandeja de plástico a temperatura ambiente (22-23 ° C). Las muestras húmedas después de la inmersión se prueban inmediatamente para la cizallaresistencia a la rotura y reportado como resistencia a la humedad. El exceso de agua en la superficie de la chapa se puede eliminar mediante golpecitos suavemente con papel tisú antes de las mediciones. </li><li> Sumerja otro conjunto de muestras de madera unidas, después del acondicionamiento inicial, en un baño de agua a 63 ºC durante 4 horas, luego seca en condiciones ambientales (temperatura de 22-23 ° C y una humedad relativa del 50 - 60%) O / N (18 - 20 h). Repita el ciclo de inmersión-secado una vez con un tiempo de secado de 48 horas. Las muestras secas se prueban para la resistencia al corte a la rotura y reportados como fuerza adhesiva empapado. </li></ol> 6. Lap resistencia al corte Mediciones <ol><li> Colocar una muestra de madera unidas en los 32 x 40 mm Fishscale mordazas de cuña reticulares en un probador de materiales con una presión de agarre de 7 MPa, y establecer la velocidad de la cruceta a 1 mm min -1. </li><li> Medir y registrar la resistencia al corte de rotura para cada muestra de madera unidas. Los resultados de las mediciones múltiples se promedian para cada forma de adhesivoulación y variable de prueba. </li></ol>

<ol>
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The authors have nothing to disclose.

Este trabajo presenta un procedimiento básico para preparar y productos como adhesivos para madera planta de prueba de semillas de base. Las lechadas adhesivas ejemplificado en este protocolo son simplemente el producto de harina de semilla desgrasada y agua. Diversas formulaciones adhesivas se pueden alcanzar mediante la adición de reactivos para las pruebas (tales como dodecilsulfato de sodio, bisulfito de sodio o de aceite de tung) 5,6,23 y / o cambios en las condiciones (tales como el pH, la proporción...

La unión adhesiva de la madera juega un papel cada vez mayor en la industria de productos forestales y es un factor clave para la utilización eficiente de los recursos maderables 1. El interés en el uso de adhesivos a base de productos naturales para la madera aumentó constantemente desde 1930 hasta alcanzar un pico alrededor de 1960 2. Después de este período, el precio de los adhesivos a base de petróleo llegó a ser tan baja que desplazaron adhesivos proteicos de varios mercados tradicionales. En las últimas dos décadas, esta tendencia se ha invertido con renovado interés en el uso de materiales que son renovables, biodegradables, y más ambientalmente aceptable. Estos recursos naturales incluyen, pero no se limitan a, proteína de soja 3-5, proteína de semilla de algodón 6, salvado de arroz 7, 8 gluten de trigo, proteína de grano de destiladores 9, de proteína de canola y aceite de 10-12, lignina de sorgo y caña de azúcar bagazo 13 , 14, y polisacáridos derivados de cáscaras de gambas 15. &lt;p class = &quot;jove_content&quot;&gt; Mientras que las semillas aislados de proteína ha sido ampliamente evaluado como adhesivos para madera potenciales, el procedimiento de aislamiento implica alcalina corrosivos y reactivos ácidos y hace que los adhesivos a base de aislar relativamente caro y menos con el medio ambiente 16. Así, algunas harinas de semillas desgrasados ​​(harinas) con o sin tratamiento también se han probado con el propósito de adhesivo, a pesar de que las propiedades adhesivas de estas comidas no se desempeñan tan bien como aislados de proteínas 17-19. Hemos fraccionado secuencialmente harina de algodón (CM) en diferentes fracciones, y examinó su fuerza adhesiva en chapas de madera pegado 20,21. La fracción sólida insoluble en agua (en lo sucesivo, de semilla de algodón se lavó comida-WCM) podría utilizarse como adhesivos para madera, comparable a la proteína de semilla de algodón aislado (CSPI), y sería menos costoso de preparar que CSPI. Fuerza de adhesión y resistencia al agua son dos parámetros críticos en la evaluación del desempeño deun material adhesivo potencial. Aquí, la fuerza adhesiva se reporta como la resistencia al corte a la rotura del enlace de vuelta de cada muestra de madera. La resistencia al agua del adhesivo se mide por el cambio en la resistencia al cizallamiento del recubrimiento de la muestra de madera unidas a la rotura debido a remojo en agua. El uso de semilla de algodón y soja comidas sin grasa como materia prima, este protocolo proporciona una manera simple y fácil de preparar y productos a base de semillas de plantas de prueba como adhesivos para madera. Este protocolo sería útil para facilitar el esfuerzo en la búsqueda de fórmulas más económicas y respetuosas del medio ambiente de los adhesivos naturales madera a base de productos.

<table border="0" cellpadding="0" cellspacing="0" width="927">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Material and Equipment</td>
			<td style="width:227px;">Supplier/Manufacturer</td>
			<td style="width:227px;"></td>
			<td style="width:211px;">Address</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;">defatted cottonseed meal</td>
			<td>Kentwood Co-op</td>
			<td></td>
			<td>Kentwood, LA, USA&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">defatted soy meal</td>
			<td>Kentwood Co-op</td>
			<td></td>
			<td>Kentwood, LA, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">wood veneers</td>
			<td>Certainly Wood, Inc.</td>
			<td></td>
			<td>East Aurora, NY, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cyclone sample mill (model 3010-014)</td>
			<td>UDY Corporation</td>
			<td></td>
			<td>Fort Collins, CO, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Benchtop Heated Press&#160; (Model 3856)</td>
			<td>Carver, Inc.</td>
			<td></td>
			<td>Wabash, IN. USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Materials Tester</td>
			<td>Zwick GmbH &#38; Co.</td>
			<td></td>
			<td>Ulm, Germany</td>
		</tr>
	</tbody>
</table>

preparation and testing of plant seed meal-based wood adhesives

Recientemente, el interés en los productos a base de harina de semilla de planta como adhesivos para madera ha aumentado de manera constante, ya que estas materias primas vegetales se consideran renovables y el medio ambiente. Estos productos naturales pueden servir como alternativas a los adhesivos a base de petróleo para aliviar las preocupaciones ambientales y de sostenibilidad. Este trabajo demuestra la preparación y ensayo de los basados ​​en semillas adhesivos para madera vegetales utilizando semillas de algodón y harina de soja como materia prima. Además de las comidas no tratados, el agua lava las comidas y aislados de proteína se preparó y ensayó. Lechadas adhesivas se preparan mezclando un producto de comida liofilizada con agua desionizada (03:25 w / w) durante 2 horas. Cada preparación de adhesivo se aplica a un extremo de tiras de chapa de madera 2 utilizando un cepillo. Las áreas de adhesivo pegajoso recubiertas de las tiras de chapa de madera se solapa engomada por prensado en caliente. Fuerza de adhesión se reporta como la resistencia al corte de la pieza de madera unidas a la rotura. La resistencia al agua de los adhesivos se mide porel cambio en la resistencia al corte de la madera en condiciones de servidumbre especímenes a la rotura después de remojo en agua. Este protocolo permite evaluar los productos agrícolas a base de semillas de plantas como candidatos adecuados para la sustitución de los adhesivos a base de madera-sintéticos. Los ajustes a la formulación de adhesivo con o sin aditivos y condiciones de unión podrían optimizar sus propiedades adhesivas para diversas aplicaciones prácticas.

El rendimiento de cada formulación de adhesivo está determinada por la resistencia al cizallamiento de la muestra de madera unidas a la rotura y los valores varían en función de las dimensiones de la chapa de madera utilizado. Por ejemplo, en la Tabla 1, los valores de resistencia adhesiva seca y se sumergen de las muestras unidas son menores cuando se utilizan tiras de arce delgado y más estrecho (ver semilla de algodón-1), en oposición a las tiras más gruesas y más amplios de semilla de algod...

Watch this Scientific Journal Video about Preparation and Testing of Plant Seed Meal-based Wood Adhesives at JoVE.com

Preparation and Testing of Plant Seed Meal-based Wood Adhesives

To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

Preparación y Pruebas de Adhesivos madera a base de comidas-Semilla de plantas

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable ...

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JoVE Core

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Civil Engineering

Environment

Unit 1

Wood

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8.6K vistas.  United States Department of Agriculture. To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

Video: Preparation and Testing of Plant Seed Meal-based Wood Adhesives

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

Un método rápido y eficiente para evaluar la patogenicidad de Ustilago maydis en las líneas de maíz y teosinte

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

Investigación

Medio ambiente

Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the exploration of renewable resources are necessary to overcome these challenges. Within the multinational framework MultiBioPro we are developing biorefinery pipelines to maximize the use of plant biomass. More specifically, we use poplar and tobacco tree (Nicotiana glauca) as target crop species for improving saccharification, isoprenoid, long chain hydrocarbon contents, fiber quality, and suberin and lignin contents. The methods used to obtain these outputs include GC-MS, LC-MS and RNA sequencing platforms. The metabolite pipelines are well established tools to generate these types of data, but also have the limitations in that only well characterized metabolites can be used. The deep sequencing will allow us to include all transcripts present during the developmental stages of the tobacco tree leaf, but has to be mapped back to the sequence of Nicotiana tabacum. With these set-ups, we aim at a basic understanding for underlying processes and at establishing an industrial framework to exploit the outcomes. In a more long term perspective, we believe that data generated here will provide means for a sustainable biorefinery process using poplar and tobacco tree as raw material. To date the basal level of metabolites in the samples have been analyzed and the protocols utilized are provided in this article.

Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline.

Transcripción y metabolito de perfiles para la Evaluación de árbol Tabaco y Poplar como materia prima para la industria basada en Bio

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the ...

A Technical Perspective in Modern Tree-ring Research - How to Overcome Dendroecological and Wood Anatomical Challenges

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to environmental changes and to finally reconstruct such changes. This is done by analyzing growth variations back in time and correlating various plant-specific parameters to (for example) temperature records. Integrating wood anatomical parameters in these analyses would strengthen reconstructions, even down to intra-annual resolution. We therefore present a protocol on how to sample, prepare, and analyze wooden specimen for common macroscopic analyses, but also for subsequent microscopic analyses. Furthermore we introduce a potential solution for analyzing digital images generated from common small and large specimens to support time-series analyses. The protocol presents the basic steps as they currently can be used. Beyond this, there is an ongoing need for the improvement of existing techniques, and development of new techniques, to record and quantify past and ongoing environmental processes. Traditional wood anatomical research needs to be expanded to include ecological information to this field of research. This would support dendro-scientists who intend to analyze new parameters and develop new methodologies to understand the short and long term effects of specific environmental factors on the anatomy of woody plants.

Here we present a protocol outlining how to sample wooden specimens for the overall assessment of their growth structures. Macro- and microscopic preparation and visualization techniques necessary to generate well-replicated and highly resolved wood anatomical and dendroecological dataset, are described are described.

Una Perspectiva Técnica en Modern Árbol-anillo de Investigación - Cómo superar dendroecológica y madera anatómicas Desafíos

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

Preparación y ensayo de impedancia basada en biochips fluido con células RTgill-W1 para evaluación rápida de muestras de agua potable para la toxicidad

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

Método fiable para evaluar la germinación de semillas, dormancia y mortalidad en condiciones de campo

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

On the Preparation and Testing of Fuel Cell Catalysts Using the Thin Film Rotating Disk Electrode Method

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a high surface area carbon, and to test their performance in thin film rotating disk electrode (TF-RDE) measurements. The TF-RDE methodology is widely used for catalyst screening; nevertheless, the measured performance sometimes considerably differs among research groups. These uncertainties impede the advancement of new catalyst materials and, consequently, several authors discussed possible best practice methods and the importance of benchmarking.
The visual tutorial highlights possible pitfalls in the TF-RDE testing of Pt/C catalysts. A synthesis and testing protocol to assess standard Pt/C catalysts is introduced that can be used together with polycrystalline Pt disks as benchmark catalysts. In particular, this study highlights how the properties of the catalyst film on the glassy carbon (GC) electrode influence the measured performance in TF-RDE testing. To obtain thin, homogeneous catalyst films, not only the catalyst preparation, but also the ink deposition and drying procedures are essential. It is demonstrated that an adjustment of the ink&#39;s pH might be necessary, and how simple control measurements can be used to check film quality. Once reproducible TF-RDE measurements are obtained, determining the Pt loading on the catalyst support (expressed as Pt wt%) and the electrochemical surface area is necessary to normalize the determined reaction rates to either surface area or Pt mass. For the surface area determination, so-called CO stripping, or the determination of the hydrogen underpotential deposition (Hupd) charge, are standard. For the determination of the Pt loading, a straightforward and cheap procedure using digestion in aqua regia with subsequent conversion of Pt(IV) to Pt(II) and UV-vis measurements is introduced.

Preparing and testing Pt/C fuel cell catalysts is subject to continuous discussion in the scientific community with respect to reproducibility and best practice. With the presented work, we intend to present a step-by-step tutorial to make and test Pt/C catalysts, which can serve as benchmark for novel catalyst systems.

En la preparación y ensayo de catalizadores de célula combustible utilizando el método de electrodo de disco giratoria de película delgada

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

La prueba de infección de planta: Spray y mediada por la herida la inoculación con el patógeno de planta Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

Xylem Water Distribution in Woody Plants Visualized with a Cryo-scanning Electron Microscope

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water distribution in plant tissues in combination with freeze fixation techniques using liquid nitrogen (LN2). For woody species, however, preparations for observing the xylem transverse-cut surface involve some difficulties due to the orientation of wood fibers. Additionally, higher tension in the water column in xylem conduits can occasionally cause artifactual changes in water distribution, especially during sample fixation and collection. In this study, we demonstrate an efficient procedure to observe the water distribution within the xylem of woody plants in situ&#160;by using a cryostat and cryo-SEM. At first, during sample collection, measuring the xylem water potential should determine whether high tension is present in the xylem conduits. When the xylem water potential is low (&#60; ca. &#8722;0.5 MPa), a tension relaxation procedure is needed to facilitate better preservation of the water status in xylem conduits during sample freeze fixation. Next, a watertight collar is attached around the tree stem and filled with LN2&#160;for freeze fixation of the water status of xylem. After harvesting, care should be taken to ensure that the sample is preserved frozen while completing the procedures of sample preparation for observation. A cryostat is employed to clearly expose the xylem transverse-cut surface. In cryo-SEM observations, time adjustment for freeze-etching is required to remove frost dust and accentuate the edge of the cell walls on the viewing surface. Our results demonstrate the applicability of cryo-SEM techniques for the observation of water distribution within xylem at cellular and subcellular levels. The combination of cryo-SEM with non-destructive in situ observation techniques will profoundly improve the exploration of woody plant water flow dynamics.

Observing the water distribution within the xylem provides significant information regarding water flow dynamics in woody plants. In this study, we demonstrate the practical approach to observe xylem water distribution in situ&#160;by using a cryostat and cryo-SEM, which eliminates artifactual changes in the water status during sample preparation.

Distribución de agua de Xylem en plantas leñosas visualizada con un microscopio electrónico de barrido crio-

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water ...

Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of the woodboring beetle, Chlorophorus caragana Xie &#38; Wang (2012), for both types of electron microscopy. The scanning electron microscopy (SEM) sample preparation protocol was based on sample chemical fixation, dehydration in a series of ethanol baths, drying, and sputter-coating. By adding Tween 20 (Polyoxyethylene sorbitan laurate) to the fixative and the wash solution, the insect body surface of woodboring beetle was washed more cleanly in SEM. This study&#39;s transmission electron microscopy (TEM) sample preparation involved a series of steps including fixation, ethanol dehydration, embedding in resin, positioning using fluorescence microscopy, sectioning, and staining. Fixative with Tween 20 enabled penetrate the insect body wall of woodboring beetle more easily than it would had been without Tween 20, and subsequently better fixed tissues and organs in the body, thus yielded clear transmission electron microscope observations of insect sensilla ultrastructures. The next step of this preparation was determining the positions of insect sensilla in the sample embedded in the resin block by using fluorescence microscopy to increase the precision of target sensilla positioning. This improved slicing accuracy.

In order to observe ultrastructure of insect sensilla, scanning and transmission electron microscopy (SEM and TEM, respectively) sample preparation protocol were presented in the study. Tween 20 was added into the fixative to avoid sample deformation in SEM. Fluorescence microscopy was helpful for improving slicing accuracy in TEM.

Método de preparación de muestras de microscopio electrónico de escaneo y transmisión para los apéndices del escarabajo carpintero

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of ...

The Effect of Construction and Demolition Waste Plastic Fractions on Wood-Polymer Composite Properties

Construction and demolition waste (CDW), including valuable materials such as plastics, have a remarkable influence on the waste sector. In order for plastic materials to be re-utilized, they must be identified and separated according to their polymer composition. In this study, the identification of these materials was performed using near-infrared spectroscopy (NIR), which identified material based on their physical-chemical properties. Advantages of the NIR method are a low environmental impact and rapid measurement (within a few seconds) in the spectral range of 1600-2400 nm without special sample preparation. Limitations include its inability to analyze dark materials. The identified polymers were utilized as a component for wood-polymer composite (WPC) that consists of a polymer matrix, low cost fillers, and additives. The components were first compounded with an agglomeration apparatus, followed by production by extrusion. In the agglomeration process, the aim was to compound all materials to produce uniformly distributed and granulated materials as pellets. During the agglomeration process, the polymer (matrix) was melted and fillers and other additives were then mixed into the melted polymer, being ready for the extrusion process. In the extrusion method, heat and shear forces were applied to a material within the barrel of a conical counter-rotating twin-screw type extruder, which reduces the risk of burning the materials and lower shear mixing. The heated and sheared mixture was then conveyed through a die to give the product the desired shape. The above-described protocol proved the potential for re-utilization of CDW materials. Functional properties must be verified according to the standardized tests, such as flexural, tensile, and impact strength tests for the material.

Secondary material streams have been shown to include potential raw materials for production. Presented here is a protocol in which CDW-plastic waste as a raw material is identified, followed by various processing steps (agglomeration, extrusion). As a result, a composite material was produced, and mechanical properties were analyzed.

El efecto de las fracciones de plástico de desecho de construcción y demolición en las propiedades compuestas de polímero de madera

Construction and demolition waste (CDW), including valuable materials such as plastics, have a remarkable influence on the waste sector. In order for ...