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. di cotone e prodotti a base di farina di soia (Figura 1) <ol><li> Ottenere le materie prime, cotone sgrassato e pasti a base di soia, dalle fonti disponibili in commercio. </li><li> Ottenere il pasto lavorando macinando pasto solido sgrassata in un mulino campione ciclone per passare una schermata di acciaio 0,5 millimetri 16. </li><li> Preparare i pasti acqua lavato dai pasti di lavoro dopo l&#39;estrazione di acqua (25 g farina 200 ml di acqua) per separare i componenti solubili in acqua nei pasti 21. </li><li> Preparare proteine ​​isolate di pasti di lavoro di estrazione alcali e precipitazioni acide 16. </li></ol> 2. Preparazione di legno impiallacciatura strisce <ol><li> Impiallacciatura di legno (1,59 millimetri di spessore) ottenuti da una fonte disponibile in commercio in strisce larghe 25,4 millimetri per 88,9 millimetri lunghi. </li><li> Matita segnare una linea attraverso le venature del legno a 25,4 mm (1,0 &quot;) di lunghezza da una estremità di ogni striscia Etichetta queste strisce appropriato con testare trattamenti o numeri 5.. -10 paia di legno sono preparati per ogni variabile test. </li></ol> 3. Preparazione di Adhesive Fanghi <ol><li> Calcolare la quantità di acqua lavato pasto necessaria per i campioni di legno per test, dal tasso di applicazione (ad esempio, 4 mg secco contenuto cm -2) x area totale di legame (ad esempio, 581 centimetri 2 su 90 listelli di legno con 2,54 x 2,54 centimetri zona di incollaggio ciascuna) più circa il 30% in più per enoughness (cioè, 4 x 581 x 130% 3 g di acqua lavato pasto per l&#39;esempio). </li><li> Mescolare acqua pasto lavato con acqua deionizzata (3:25 w / w), e mescolare con un bastoncino magnetico per 2 ore in un bicchiere sigillato con Parafilm. </li></ol> 4. Preparazione di Bonded legno campioni <ol><li> Spazzolare liquami adesivo su una delle estremità delle strisce impiallacciatura di legno che coprono 2 25,4 mm (1,0 &quot;) di lunghezza di aria secca per 10 - 15 min. O fino cattivo gusto. </li><li> Spazzola un secondo strato di boiacca adesiva sulla parte superiore del primo strato e asciugare di nuovo. La quantità di adesivo seccoPreparazione applicato è di circa 4,5 mg secco solido per cm 2 di incollaggio sono di ogni striscia di legno. </li><li> Sovrapporre l&#39;area rivestita adesiva appiccicoso (25,4 x 25,4 millimetri o 1,0 &quot;x 1,0&quot;) di strisce di impiallacciatura di legno 2. Hot-press utilizzando un banco riscaldata Premere a 100 ° C per 20 minuti ad una pressione di 400 psi (2,8 MPa). Nota la pressione è la forza applicata dalla stampa diviso per l&#39;area di sovrapposizione dei campioni di legno. Questi parametri di legame possono essere modificati come necessario per ciascuna variabile test. </li><li> Raffreddare e condizionare i campioni di legno legati per 48 ore in una camera di condizionamento o di un incubatore con controllo dell&#39;umidità (temperatura di 22 - 23 ° C ed umidità relativa del 50 - 60%; la figura 2). </li></ol> 5. resistenza all&#39;acqua Experiments <ol><li> Immergere i campioni di legno incollati, dopo condizionamento iniziale, in acqua corrente per 48 h in un vassoio di plastica a RT (22 - 23 ° C). Gli esemplari bagnato dopo ammollo sono testati immediatamente per il taglioresistenza alla rottura e segnalato come resistenza all&#39;acqua. L&#39;acqua in eccesso sulla superficie rivestimento può essere rimosso tamponando delicatamente con carta velina prima di misurazioni. </li><li> Immergere un&#39;altra serie di campioni di legno incollati, dopo condizionamento iniziale, in un bagno di acqua a 63 ° C per 4 ore, poi si essicca a condizioni ambientali (temperatura di 22 - 23 ° C ed umidità relativa del 50 - 60%) O / N (18 - 20 ore). Ripetere il ciclo di immersione asciugatura volta con un tempo di asciugatura 48 hr. I campioni essiccati vengono poi testati per la resistenza al taglio in pausa e riportati come forza adesiva imbevuto. </li></ol> 6. Misure Lap Shear Forza <ol><li> Montare un campione di legno incollate nelle Scaglie di pesce morsetti a cuneo reticolati 32 x 40 mm su un Tester Materials con una pressione di presa di 7 MPa, e impostare la velocità della traversa a 1 mm min -1. </li><li> Misurare e registrare la resistenza al taglio a rottura per ogni campione di legno incollati. I risultati delle misurazioni multiple vengono mediate per ciascuna forma adesivalamento e variabile del test. </li></ol>

<ol>
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The authors have nothing to disclose.

Questo articolo presenta una procedura di base per la preparazione e prodotti come adesivi di legno vegetale seed test-based. I fanghi adesivi exampled in questo protocollo sono semplicemente il prodotto farina di semi scremato e dell&#39;acqua. Varie formulazioni adesive possono essere raggiunti mediante aggiunta di reagenti di prova (come dodecil solfato di sodio, bisolfito di sodio o olio tung) 5,6,23 e / o variazioni di condizioni (quali pH, rapporto tra solido e acqua) miscelazione 3,24 , 25. ...

Incollaggio di legno gioca un ruolo sempre più importante nel settore dei prodotti della foresta ed è un fattore chiave per l&#39;utilizzo efficiente delle risorse di legname 1. L&#39;interesse per l&#39;uso di adesivi a base naturale dei prodotti del legno è aumentato costantemente dal 1930 a raggiungere un picco intorno al 1960 2. Dopo questo periodo, il prezzo di adesivi a base di petrolio è diventato così basso che sfollate adesivi proteine ​​da diversi mercati tradizionali. Negli ultimi due decenni, questa tendenza è invertita con rinnovato interesse per l&#39;utilizzo di materiali che sono rinnovabili, biodegradabili, e più ambientalmente accettabile. Queste risorse naturali comprendono, ma non sono limitati a, proteine ​​di soia 3-5, proteine ​​di cotone 6, crusca di riso 7, glutine di frumento 8, proteine ​​del grano distillatori 9, proteine ​​colza e olio 10-12, lignina da sorgo e di canna da zucchero bagasse 13 , 14, e polisaccaridi derivati ​​da gusci di gamberetti 15. &lt;p class = &quot;jove_content&quot;&gt; considerando che gli isolati di proteine ​​di semi sono stati ampiamente valutati come potenziali adesivi per legno, la procedura di isolamento comporta alcalina corrosivo e reagenti acidi e rende adesivi isolare basata relativamente costoso e meno favorevole all&#39;ambiente 16. Così, alcuni pasti seme sgrassate (farine) con o senza trattamento sono stati anche testati per fini adesivo, anche se le proprietà adesive di questi pasti non eseguono nonché proteine ​​isolate 17-19. Abbiamo sequenza frazionata pasto di semi di cotone (CM) in diverse frazioni, e ha esaminato la loro forza adesiva in impiallacciature di legno bonding 20,21. La frazione solida insolubile in acqua (cotone lavato seguito pasto-WCM) potrebbe essere usato come adesivi per legno, paragonabili alle proteine ​​di cotone isolato (CSPI), e sarebbe meno costosa per la preparazione di CSPI. Forza adesiva e resistenza all&#39;acqua sono due parametri critici per valutare le prestazioni diun potenziale materiale adesivo. Qui, la forza adesiva è riportato come la resistenza al taglio a rottura del legame giro di ogni campione di legno. Resistenza all&#39;acqua dell&#39;adesivo viene misurato dalla variazione nella resistenza al taglio per trazione del campione di legno legati a rottura a causa di ammollo acqua. Utilizzando sgrassate di cotone e soia pasti come materie prime, questo protocollo fornisce un modo semplice e diretto per preparare e vegetali di prova prodotti a base di semi-come adesivi legno. Questo protocollo sarebbe utile nel facilitare lo sforzo nella ricerca di formulazioni più economiche e rispettose dell&#39;ambiente di adesivi naturali del legno a base di prodotti.

<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

Recentemente, l&#39;interesse nelle sementi di piante a base pasto-prodotti come adesivi per legno è costantemente aumentato, come queste materie prime vegetali sono considerati rinnovabile e rispettosa dell&#39;ambiente. Questi prodotti naturali possono servire come alternative ai adesivi a base di petrolio per facilitare le preoccupazioni ambientali e di sostenibilità. Questo lavoro dimostra la preparazione e collaudo degli adesivi a base di legno di semi-vegetale con cotone e farina di soia come materie prime. In aggiunta ai pasti non trattati, acqua lavato piatti e isolati proteici sono preparati e testati. Boiacche adesive vengono preparati miscelando un pasto prodotto liofilizzato con acqua deionizzata (03:25 w / w) per 2 ore. Ogni preparazione adesivo è applicato ad una estremità di strisce di impiallacciatura 2 legno utilizzando un pennello. Le aree adesivo viscoso rivestite delle strisce impiallacciatura di legno sono falde incollate da pressatura a caldo. Forza adesiva è segnalato come la resistenza al taglio del campione di legno incollate a rottura. Resistenza all&#39;acqua degli adesivi è misurato dalil cambiamento di resistenza a taglio del legno incollato provini a rottura dopo ammollo acqua. Questo protocollo permette di valutare i prodotti agricoli a base di semi di piante come candidati idonei per la sostituzione di adesivi a base di legno sintetiche. Adeguamenti formulazione adesiva con o senza additivi e condizioni di incollaggio possono ottimizzare le loro proprietà adesive per diverse applicazioni pratiche.

Il rendimento di ciascuna formulazione adesiva è determinata dalla resistenza al taglio del campione di legno legati a rottura e valori variano a seconda delle dimensioni della impiallacciatura di legno utilizzato. Per esempio, nella tabella 1, i valori di resistenza adesiva asciutti e bagnati dei provini incollati sono inferiori quando si utilizzano nastri di acero sottili e stretti (vedi cotone-1), in contrasto con le strisce più spesse e più ampia di cotone-2 raccomandate nel protocollo, utilizzan...

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.

Preparazione e test di adesivi a base di legno pasto pianta Seed

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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8.6K Views.  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 metodo rapido ed efficiente per valutare la patogenicità dei maydis di Ustilago sulle linee di mais e teosinte

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

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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.

Trascrizione e metabolita Profiling per la valutazione del Tabacco Albero e pioppo come materia prima per l&#39;industria con sede a 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.

Un punto di vista tecnico in Modern Tree-Ring Research - Come superare Dendroecological e legno anatomiche sfide

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.

Preparazione e test di impedenza a base di biochip fluidici con le cellule RTgill-W1 per valutare rapidamente campioni di acqua potabile per la tossicità

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.

Metodo affidabile per valutare la germinazione dei semi, dormienza, e mortalità in normali condizioni operative

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.

Preparazione e test di catalizzatori di celle a combustibile che utilizza il metodo di elettrodo disco rotante a Film sottile

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 prova di infezione di pianta: Spray e ferita-mediata di inoculazione con la pianta patogeno 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.

Distribuzione dell'acqua Xylem nelle piante legnose con un microscopio elettronico a scansione crio-scansione

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.

Esempio di metodo di preparazione del microscopio elettronico di scansione e trasmissione per le appendici di Woodboring Beetle

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.

L'effetto delle frazioni di plastica dei rifiuti di costruzione e demolizione sulle proprietà composite wood-Polymer

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