The research was supported by the National Research Council (CNR) of Italy. The authors thank GIRE members for collecting seed samples and are grateful to Alison Garside for revising the English.

<ol>
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J., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target-site+resistance+to+ALS+inhibitors+in+the+polyploid+species+Echinochloa+crus-galli.">Target-site resistance to ALS inhibitors in the polyploid species Echinochloa crus-galli.</a> Pestic. Biochem. Phys. 105 (2), 93-101 (2013).</li><li>Sattin, M., Berto, D., Zanin, G., Tabacchi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+to+ALS+inhibitors+in+rice+in+north-western+Italy.">Resistance to ALS inhibitors in rice in north-western Italy.</a> Proc. BCPC. Weeds. , 783-790 (1999).</li><li>Scarabel, L., Berto, D., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dormancy+breaking+and+germination+of+Alisma+plantago-aquatica.+and+Scirpus+mucronatus.">Dormancy breaking and germination of Alisma plantago-aquatica. and Scirpus mucronatus.</a> Aspects of Applied Biology. 69, 285-292 (2003).</li><li>Collavo, A., Strek, H., Beffa, R., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+an+ACCase-inhibitor-resistant+Lolium+rigidum.+population+based+on+the+use+of+ALS+inhibitors:+weed+population+evolution+observed+over+a+7+years+field-scale+investigation.">Management of an ACCase-inhibitor-resistant Lolium rigidum. population based on the use of ALS inhibitors: weed population evolution observed over a 7 years field-scale investigation.</a> Pest Manag. Sci. 69 (2), 200-208 (2013).</li><li>Scarabel, L., Panozzo, S., Savoia, W., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target-site+ACCase-resistant+Johnsongrass+(Sorghum+halepense).+selected+in+summer+dicot+crops.">Target-site ACCase-resistant Johnsongrass (Sorghum halepense). selected in summer dicot crops.</a> Weed Technol. 28 (2), 307-315 (2014).</li><li>Hess, M., Barralis, H., Bleiholder, H., Buhur, L., Eggers, T., Hack, H., Strauss, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+the+extended+BBCH+scale+-+general+for+the+description+of+the+growth+stages+of+mono-+and+dicotyledonous+weed+species.">Use of the extended BBCH scale - general for the description of the growth stages of mono- and dicotyledonous weed species.</a> Weed Res. 37 (6), 433-441 (1997).</li><li>Collavo, A., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+glyphosate-resistant+Lolium.+spp.+biotypes+found+in+a+European+annual+arable+cropping+system+also+affected+by+ACCase+and+ALS+resistance.">First glyphosate-resistant Lolium. spp. biotypes found in a European annual arable cropping system also affected by ACCase and ALS resistance.</a> Weed Res. 54 (4), 325-334 (2014).</li><li>Scarabel, L., Cenghialta, C., Manuello, D., Sattin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+and+management+of+imidazolinone-resistant+red+rice+(Oryza+sativa.+L.,+var.+sylvatica.)+in+Clearfield&#174;+Italian+paddy+rice.">Monitoring and management of imidazolinone-resistant red rice (Oryza sativa. L., var. sylvatica.) in Clearfield&#174; Italian paddy rice.</a> Agronomy. 2 (4), 371-383 (2012).</li><li>Zelaya, I. A., Anderson, J. A. H., Owen, M. D. K., Landes, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+spectrophotometric+and+HPLC+methods+for+shikimic+acid+determination+in+plants:+Models+in+glyphosate-resistant+and-susceptible+crops.">Evaluation of spectrophotometric and HPLC methods for shikimic acid determination in plants: Models in glyphosate-resistant and-susceptible crops.</a> J. Agric. Food Chem. 59 (6), 2202-2212 (2011).</li></ol>

The authors declare that they have no competing financial interests.

כמה צעדים בפרוטוקולים הם קריטיים להערכה מוצלחת של התנגדות קוטל עשבים באוכלוסייה: 1) יש לאסוף זרעים כאשר בוגר מצמחים ששרדו את טיפול קוטל עשבים (ים). הבשלה של הזרעים על הצמח האם הוא חיוני כדי למנוע קשיים בנביטת זרעים מאוחר יותר; 2) האחסון הנכון של זרעים מומלץ להימנע התפשט?...

<html> קוטלי עשבים הם מידת שליטת העשב ביותר בשימוש נרחב, והיוו עד 50% משוק הגנת הצומח העולמי 1. הם כלים זולים יחסית, להימנע משיטות טיפוח אדמה עתירת עבודה וזמן רב, וסופו של דבר להביא לייצור חסכוני, בטוח ורווחי מזון 2. עם זאת, המתנה הגדולה phenological והגנטי השונות במינים עשבים רבים, יחד עם הסתמכות יתר על שימוש קוטל עשבים, לעתים קרובות גורמת לבחירה של אוכלוסיות עשבים עמידים בפני קוטלי עשבים. המבוא של קוטלי עשבים סלקטיביים עם יעד חילוף חומרים מאוד ספציפי 3-5 כבר עלה באופן משמעותי את מספר המקרים התנגדות לאורך השנים. עד כה, 240 מיני עשבים (140 dicots וmonocots 100) ברחבי העולם התפתחו התנגדות לאתרים שונים של קוטל עשבים פעולה (SOA) 4. זוהי דאגה גדולה לניהול עשב ויותר באופן כללי לייצור יבול בר-קיימא. e_content &quot;&gt; גילוי מוקדם של התנגדות, המבוסס על בדיקות אמינות, מבוצעות לעתים קרובות בחממה, הוא צעד מפתח לנהל עשבים עמידים קוטל עשבים. גישות שונות פותחו בהתאם למטרות, הרמה הנדרשת של דיוק, זמן ומשאבים זמינים, כ גם מיני עשבים נחשבים 6-12. עם זאת, כאשר אישור של מעמד ההתנגדות של biotype עשב חדש נדרש (כלומר, קבוצה של אנשים שחולקות כמה מאפיינים פיסיולוגיים, כולל היכולת לשרוד אחד או יותר קוטלי עשבים השייכים ל קבוצה מסוימת בשימוש במינון שהיה בדרך כלל לשלוט בם), המבדק צמח שלם חזק צריכה להיות מבוצעת בסביבה מבוקרת 4, 11. Biotype הוא לעתים רחוקות עמיד לקוטל עשבים רק אחד. כל biotype מתאפיין לכן בדפוס מסוים התנגדות, מספר כלומר, והסוג של SOA של קוטלי העשבים הוא עמיד ל, ועל ידי התנגדות ניתנהרמה לכל קוטלי עשבים 13. הקביעה המוקדמת ואמינה של הדפוס של צלב או התנגדות מרובה 5, 14 היא חשובה לניהול התנגדות שדה. ראוי להזכיר כי התנגדות קוטל עשבים אין שום קשר עם הסובלנות הטבעית שכמה מיני תערוכת עשב כלפי כמה קוטלי עשבים, למשל, מיני dicot לעומת קוטלי עשבים ACCase-עיכוב, מיני monocot לעומת 2,4-D, Equisetum arvense לעומת גלייפוסט. מאמר זה מציג גישה חזקה לבדיקת biotypes עמיד קוטל עשבים משוערים שנדגמו בתחומים שבם שליטה עניה על ידי קוטל עשבים (ים) שדווחו. גרסאות רלוונטיות לפרוטוקולים סטנדרטיים ביחס למיני העשבים המעורבים מוצגות. היתרונות על פני שיטות / פרוטוקולים חלופיים המבוססים על שני bioassays הצמח כולו רק באמצעות מנה אחת קוטל עשבים 15, או זרעי טיפול בצלחות פטרי 8 קשורים לreliab גבוה יותרility והאפשרות להסיק רמת ההתנגדות בגלל ההכללה של שני מינוני קוטל עשבים בניסויים. עם זאת, לבדיקת התנגדות שגרתית, יכולות להיות מיושמות באותן שיטות וברק מנה אחת קוטל עשבים, ולכן הפחתת העלויות. כמו גם לאפשר אישור של מעמד ההתנגדות, המידע שהושג יכול לשמש לייעול פעולות המחקר הבא ו / או להמציא אסטרטגיות ניהול קול התנגדות.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Paper bags</td>
			<td>Celcar SAS</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic dishes</td>
			<td>ISI plast S.p.A.</td>
			<td>SO600</td>
			<td>Transparent plastic</td>
		</tr>
		<tr>
			<td>Sulfuric acid 95-98%</td>
			<td>Sigma-Aldrich</td>
			<td>320501</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-woven fabric</td>
			<td>Carretta Tessitura</td>
			<td>Art.TNT17</td>
			<td>Weight&#160; 17 gr m-&#178;</td>
		</tr>
		<tr>
			<td>Chloroform &#62;99.5%</td>
			<td>Sigma-Aldrich</td>
			<td>C2432</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>Sigma-Aldrich</td>
			<td>A1296</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium nitrate &#62;99.0%</td>
			<td>Sigma-Aldrich</td>
			<td>P8394</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic containers</td>
			<td>Giganplast</td>
			<td>1875/M</td>
			<td>600 x 400 x 110 mm</td>
		</tr>
		<tr>
			<td>Plastic trays</td>
			<td>Piber plast</td>
			<td>G1210A</td>
			<td>325 x 265 x 95 mm</td>
		</tr>
		<tr>
			<td>Polystyrene trays</td>
			<td>Plastisavio</td>
			<td>S24</td>
			<td>537x328x72 mm, 24 round cells (6x4)&#160;</td>
		</tr>
		<tr>
			<td>Copper sulfate</td>
			<td>Sigma-Aldrich</td>
			<td>451657</td>
			<td></td>
		</tr>
		<tr>
			<td>Agriperlite</td>
			<td>Blu Agroingross sas</td>
			<td>AGRI100</td>
			<td></td>
		</tr>
		<tr>
			<td>Peat</td>
			<td>Blu Agroingross sas</td>
			<td>TORBA250</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Germination cabinet</td>
			<td>KW</td>
			<td>W87R</td>
			<td></td>
		</tr>
		<tr>
			<td>Nozzles</td>
			<td>Teejet</td>
			<td>&#160;XR11002-VK, TP11001-VH</td>
			<td>The second type of nozzles are used only for glyphosate</td>
		</tr>
		<tr>
			<td>Barcode generator</td>
			<td>Toshiba TEC</td>
			<td>SX4</td>
			<td></td>
		</tr>
		<tr>
			<td>Labels with barcode</td>
			<td>Felga</td>
			<td>TT20200</td>
			<td>Stick-in labels with rounded corners</td>
		</tr>
		<tr>
			<td>Barcode reader</td>
			<td>Cipherlab</td>
			<td>8300-L</td>
			<td>Portable data terminal</td>
		</tr>
		<tr>
			<td>Bench sprayer</td>
			<td>-</td>
			<td>-</td>
			<td>Built in house</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td colspan="2">HERBICIDES INCLUDED IN THE RESULTS:</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Commercial product</td>
			<td>Active ingredient</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Altorex</td>
			<td>imazamox</td>
			<td>BASF</td>
			<td></td>
		</tr>
		<tr>
			<td>Azimut&#160;</td>
			<td>florasulam</td>
			<td>Dow AgroSciences</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopower</td>
			<td></td>
			<td>Bayer Crop Science</td>
			<td>Surfact to be used with Hussar WG</td>
		</tr>
		<tr>
			<td>Dash</td>
			<td></td>
			<td>BASF</td>
			<td>Surfact to be used with Altorex</td>
		</tr>
		<tr>
			<td>Granstar&#160;</td>
			<td>tribenuron-methyl</td>
			<td>Dupont</td>
			<td></td>
		</tr>
		<tr>
			<td>Gulliver&#160;</td>
			<td>azimsulfuron</td>
			<td>Dupont</td>
			<td></td>
		</tr>
		<tr>
			<td>Hussar WG&#160;</td>
			<td>iodosulfuron</td>
			<td>Bayer Crop Science</td>
			<td></td>
		</tr>
		<tr>
			<td>Nominee&#160;</td>
			<td>bispyribac-Na</td>
			<td>Bayer Crop Science</td>
			<td></td>
		</tr>
		<tr>
			<td>Roundup</td>
			<td>glyphosate</td>
			<td>Monsanto</td>
			<td></td>
		</tr>
		<tr>
			<td>Trend</td>
			<td></td>
			<td>Dupont</td>
			<td>Surfact to be used with Granstar and Gulliver</td>
		</tr>
		<tr>
			<td>Viper&#160;</td>
			<td>penoxsulam</td>
			<td>Dow AgroSciences</td>
			<td></td>
		</tr>
		<tr>
			<td>Weedone LV4</td>
			<td>2,4-D</td>
			<td>Isagro</td>
			<td></td>
		</tr>
	</tbody>
</table>

protocols for robust herbicide resistance testing in different weed species

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented protocols, based on whole-plant bioassays performed in a greenhouse, can be readily adapted to a wide range of weed species and herbicides through appropriate variants. Seed samples from plants that survived a field herbicide treatment are collected and stored dry at low temperature until used. Germination methods differ according to weed species and seed dormancy type. Seedlings at similar growth stage are transplanted and maintained in the greenhouse under appropriate conditions until plants have reached the right growth stage for herbicide treatment. Accuracy is required to prepare the herbicide solution to avoid unverifiable mistakes. Other critical steps such as the application volume and spray speed are also evaluated. The advantages of this protocol, compared to others based on whole plant bioassays using one herbicide dose, are related to the higher reliability and the possibility of inferring the resistance level. Quicker and less expensive in vivo or in vitro diagnostic screening tests have been proposed (Petri dish bioassays, spectrophotometric tests), but they provide only qualitative information and their widespread use is hindered by the laborious set-up that some species may require. For routine resistance testing, the proposed whole plant bioassay can be applied at only one herbicide dose, so reducing the costs.

כדי להעריך את המצב של התנגדות אוכלוסייה עמידה משוערת, זה הוא יסוד לכולל בדיקה רגישה בassay על מנת לוודא את יעילות קוטלי העשבים. התוצאות של בדיקת סקר שנערכו על פ אוכלוסיות rhoeas, עשב שפשט על שדות חיטה, מדווחות באיור 2, שבו היעילות של ארבעה קוטלי עשבים הודע?...

Watch this Scientific Journal Video about Protocols for Robust Herbicide Resistance Testing in Different Weed Species at JoVE.com

Protocols for Robust Herbicide Resistance Testing in Different Weed Species

A robust and flexible approach to confirm herbicide resistance in weed populations is presented. This protocol allows the herbicide resistance levels to be inferred and applied to a wide range of weed species and herbicides with minor adaptations.

פרוטוקולים לקוטל עשבים חזקים התנגדות בדיקה במיני עשבים שונים

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented ...

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14.4K Views.  National Research Council (CNR), Italy. A robust and flexible approach to confirm herbicide resistance in weed populations is presented. This protocol allows the herbicide resistance levels to be inferred and applied to a wide range of weed species and herbicides with minor adaptations.

Video: Protocols for Robust Herbicide Resistance Testing in Different Weed Species

An Optimized Enrichment Technique for the Isolation of Arthrobacter Bacteriophage Species from Soil Sample Isolates

Bacteriophage isolation from environmental samples has been performed for decades using principles set forth by pioneers in microbiology. The isolation of phages infecting Arthrobacter hosts has been limited, perhaps due to the low success rate of many previous isolation techniques, resulting in an underrepresented group of Arthrobacter phages available for study. The enrichment technique described here, unlike many others, uses a filtered extract free of contaminating bacteria as the base for indicator bacteria growth, Arthrobactersp. KY3901, specifically. By first removing soil bacteria the target phages are not hindered by competition with native soil bacteria present in initial soil samples. This enrichment method has resulted in dozens of unique phages from several different soil types and even produced different types of phages from the same enriched soil sample isolate. The use of this procedure can be expanded to most nutrient rich aerobic media for the isolation of phages in a vast diversity of interesting host bacteria.

An Optimized Enrichment Technique for the Isolation of Arthrobacter Bacteriophage Species from Soil Sample Isolates

We present an enrichment protocol for the isolation of bacteriophages infecting bacteria in the Arthrobacter genus. This enrichment protocol produces fast and reproducible results for the isolation and amplification of Arthrobacter phages from soil isolates.

טכניקת העשרה אופטימלית עבור הבידוד של<em&gt; Arthrobacter</em&gt; Bacteriophage מינים מדגימת הקרקע מבודדת

Bacteriophage isolation from environmental samples has been performed for decades using principles set forth by pioneers in microbiology. The isolation of ...

Measuring Rates of Herbicide Metabolism in Dicot Weeds with an Excised Leaf Assay

In order to isolate and accurately determine rates of herbicide metabolism in an obligate-outcrossing dicot weed, waterhemp (Amaranthus tuberculatus), we developed an excised leaf assay combined with a vegetative cloning strategy to normalize herbicide uptake and remove translocation as contributing factors in herbicide-resistant (R) and &#8211;sensitive (S) waterhemp populations. Biokinetic analyses of organic pesticides in plants typically include the determination of uptake, translocation (delivery to the target site), metabolic fate, and interactions with the target site. Herbicide metabolism is an important parameter to measure in herbicide-resistant weeds and herbicide-tolerant crops, and is typically accomplished with whole-plant tests using radiolabeled herbicides. However, one difficulty with interpreting biokinetic parameters derived from whole-plant methods is that translocation is often affected by rates of herbicide metabolism, since polar metabolites are usually not mobile within the plant following herbicide detoxification reactions. Advantages of the protocol described in this manuscript include reproducible, accurate, and rapid determination of herbicide degradation rates in R and S populations, a substantial decrease in the amount of radiolabeled herbicide consumed, a large reduction in radiolabeled plant materials requiring further handling and disposal, and the ability to perform radiolabeled herbicide experiments in the lab or growth chamber instead of a greenhouse. As herbicide resistance continues to develop and spread in dicot weed populations worldwide, the excised leaf assay method developed and described herein will provide an invaluable technique for investigating non-target site-based resistance due to enhanced rates of herbicide metabolism and detoxification.

This manuscript describes how herbicide metabolism rates can be effectively quantified with excised leaves from a dicot weed, thereby reducing variability and removing any possible confounding effects of herbicide uptake or translocation typically observed in whole-plant assays.

מדידת מחירים של קוטל העשבים מטבוליזם בעשבי Dicot עם Assay נכרת עלה

In order to isolate and accurately determine rates of herbicide metabolism in an obligate-outcrossing dicot weed, waterhemp (Amaranthus tuberculatus), we ...

Potentiodynamic Corrosion Testing

Different metallic materials have different polarization characteristics as dictated by the open circuit potential, breakdown potential, and passivation potential of the material. The detection of these electrochemical parameters identifies the corrosion factors of a material. A reliable and well-functioning corrosion system is required to achieve this.
Corrosion of the samples was achieved via a potentiodynamic polarization technique employing a three-electrode configuration, consisting of reference, counter, and working electrodes. Prior to commencement a baseline potential is obtained. Following the stabilization of the corrosion potential (Ecorr), the applied potential is ramped at a slow rate in the positive direction relative to the reference electrode. The working electrode was a stainless steel screw. The reference electrode was a standard Ag/AgCl. The counter electrode used was a platinum mesh. Having a reliable and well-functioning in vitro corrosion system to test biomaterials provides an in-expensive technique that allows for the systematic characterization of the material by determining the breakdown potential, to further understand the material's response to corrosion. The goal of the protocol is to set up and run an in vitro potentiodynamic corrosion system to analyze pitting corrosion for small metallic medical devices.

Here, we present a protocol to set up and run an in vitro potentiodynamic corrosion system to analyze pitting corrosion for small metallic medical devices.

Potentiodynamic בדיקת קורוזיה

Different metallic materials have different polarization characteristics as dictated by the open circuit potential, breakdown potential, and passivation ...

Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes onto humans. For this reason, transferable pesticide residue experiments are required for registration and re-registration by the United States Environmental Protection Agency (USEPA). Although such experiments are required, limited specificity is required pertaining to experimental approach. Experimental approaches used to assess pesticide transfer to humans including hand wiping with cotton gloves, modified California roller (moving a roller of known mass over cotton cloth) and soccer ball roll (ball wrapped with sorbent strip) over three treated turfgrass species (creeping bentgrass, hybrid bermudagrass and tall fescue maintained at 0.4, 5 and 9 cm, respectively) are presented. The modified California roller is the most extensively utilized approach to date, and is best suited for use at low mowing heights due to its reproducibility and large sampling area. The soccer ball roll is a less aggressive transfer approach; however, it mimics a very common occurrence in the most popular international sport, and has many implications for nondietary pesticide exposure from hand-to-mouth contact. Additionally, this approach may be adjusted for other athletic activities with limited modification. Hand wiping is the best approach to transfer pesticides at higher mowing heights, as roller-based approaches can lay blades over; however, it is more subjective due to more variable sampling pressure. Utility of these methods across turfgrass species is presented, and additional considerations to conduct transferable pesticide residue research in turfgrass systems are discussed.

Transferable pesticide residue experiments in turfgrass systems are integral components of human risk exposure assessments. Experimental approaches to measure transferable residues should be adjusted to the human interaction of interest and turfgrass system dynamics. Three transferable pesticide residue protocols are presented and the suitability across three turfgrass systems is discussed.

פרוטוקולים לכימות שאריות חומר הדברה להעברת מערכות Turfgrass

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes ...

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and stability of the suspension. In this study, a systematic step-wise approach is carried out to identify optimal sonication conditions in order to achieve a stable dispersion. This approach has been adopted and shown to be suitable for several nanomaterials (cerium oxide, zinc oxide, and carbon nanotubes) dispersed in deionized (DI) water. However, with any change in either the nanomaterial type or dispersing medium, there needs to be optimization of the basic protocol by adjusting various factors such as sonication time, power, and sonicator type as well as temperature rise during the process. The approach records the dispersion process in detail. This is necessary to identify the time points as well as other above-mentioned conditions during the sonication process in which there may be undesirable changes, such as damage to the particle surface thus affecting surface properties. Our goal is to offer a harmonized approach that can control the quality of the final, produced dispersion. Such a guideline is instrumental in ensuring dispersion quality repeatability in the nanoscience community, particularly in the field of nanotoxicology.

Here, we present a step-wise protocol for the dispersion of nanomaterials in aqueous media with real-time characterization to identify the optimal sonication conditions, intensity, and duration for improved stability and uniformity of nanoparticle dispersions without impacting the sample integrity.

פיזור של ננו בתקשורת מימית: לקראת פרוטוקול אופטימיזציה

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and ...

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop cultivars against environmental stresses. An experimental protocol is presented for RGB and hyperspectral imaging of root systems. The approach uses rhizoboxes where plants grow in natural soil over a longer time to observe fully developed root systems. Experimental settings are exemplified for assessing rhizobox plants under water stress and studying the role of roots. An RGB imaging setup is described for cheap and quick quantification of root development over time. Hyperspectral imaging improves root segmentation from the soil background compared to RGB color based thresholding. The particular strength of hyperspectral imaging is the acquisition of chemometric information on the root-soil system for functional understanding. This is demonstrated with high resolution water content mapping. Spectral imaging however is more complex in image acquisition, processing and analysis compared to the RGB approach. A combination of both methods can optimize a comprehensive assessment of the root system. Application examples integrating root and aboveground traits are given for the context of plant phenotyping and plant physiological research. Further improvement of root imaging can be obtained by optimizing RGB image quality with better illumination using different light sources and by extension of image analysis methods to infer on root zone properties from spectral data.

An experimental protocol is presented for assessment of soil grown plant root systems with RGB and hyperspectral imaging. Combination of RGB image time series with chemometric information from hyperspectral scans optimizes insights into plant root dynamics.

RGB והדמיה ספקטרלי שורש הצמח Phenotyping ומחקר פיזיולוגיים: הגדרת הניסוי והדמיה פרוטוקולים

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A better mechanistic understanding of these complex plant-microbe interactions will be crucial to improving plant production as well as performing basic ecological studies investigating plant-soil-microbe interactions. Here, a detailed description for ecosystem fabrication is presented, using widely available 3D printing technologies, to create controlled laboratory habitats (EcoFABs) for mechanistic studies of plant-microbe interactions within specific environmental conditions. Two sizes of EcoFABs are described that are suited for the investigation of microbial interactions with various plant species, including Arabidopsis thaliana, Brachypodium distachyon, and Panicum virgatum. These flow-through devices allow for controlled manipulation and sampling of root microbiomes, root chemistry as well as imaging of root morphology and microbial localization. This protocol includes the details for maintaining sterile conditions inside EcoFABs and mounting independent LED light systems onto EcoFABs. Detailed methods for addition of different forms of media, including soils, sand, and liquid growth media coupled to the characterization of these systems using imaging and metabolomics are described. Together, these systems enable dynamic and detailed investigation of plant and plant-microbial consortia including the manipulation of microbiome composition (including mutants), the monitoring of plant growth, root morphology, exudate composition, and microbial localization under controlled environmental conditions. We anticipate that these detailed protocols will serve as an important starting point for other researchers, ideally helping create standardized experimental systems for investigating plant-microbe interactions.

Ecosystem Fabrication EcoFAB Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

This article describes detailed protocols for ecosystem fabrication of devices (EcoFABs) that enable the studies of plants and plant-microbe interactions in highly controlled laboratory conditions.

המערכת האקולוגית פרוטוקולים פבריקציה נוספת (EcoFAB) להקמת מערכות אקולוגיות מעבדה שנועדה ללמוד אינטראקציות צמח-חיידק

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

הלחנה וניתוח הפצה Bioaerosols בתנאים סביבתיים שונים

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the local to federal levels. The continuing evolution of PFAS chemistries presents a challenge to the environmental monitoring, where ongoing development of targeted methods necessarily lags the discovery of new chemical compounds. There is a need, therefore, to have forward-looking methodologies that can detect emerging and unexpected compounds, monitor these species over time, and resolve details of their chemical structure to enable future work in human health. To this end, non-targeted analysis by high-resolution mass spectrometry offers a broad base detection approach that can be combined with almost any sample preparation scheme and provides significant capabilities for compound identification after detection. Herein, we describe a solid-phase extraction (SPE) based sample concentration method tuned for shorter chain and more hydrophilic PFAS chemistries, such as per fluorinated ether acids and sulfonates, and describe analysis of samples prepared in this fashion in both targeted and non-targeted modes. Targeted methods provide superior quantification when reference standards are available but are intrinsically limited to expected compounds when performing analysis. In contrast, a non-targeted approach can identify the presence of unexpected compounds and provide some information about their chemical structure. Information about chemical features can be used to correlate compounds across sample locations and track abundance and occurrence over time.

Here, we present a protocol for the sequential targeted quantification and non-targeted analysis of fluorinated compounds in water by mass spectrometry. This methodology provides quantitative levels of known fluorochemical compounds and identifies unknown chemicals in related samples with semi-quantitative estimates of their abundance.

זיהוי לכל-כימית Polyfluorinated מינים בזרימת עבודה משולבת יישוב, שאינו ממוקד-סינון ברזולוציה גבוהה ספקטרומטר מסה

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the ...

Sampling for Estimating Frankliniella Species Flower Thrips and Orius Species Predators in Field Experiments

The western flower thrips, Frankliniella occidentalis (Pergande), is a polyphagous pest that has been spread worldwide. The extensive use of insecticides in attempts to control its populations eliminates natural enemies and competitor flower thrips species, thereby increasing its populations. An unsustainable situation develops with concomitant resistant pest populations, secondary pest outbreaks, and environmental degradation. Integrated pest management utilizes knowledge of pest and natural enemy relationships to implement tactics that are environmentally friendly and sustainable. Minute pirate bugs are the most important worldwide predators of thrips. They can suppress and ultimately control Frankliniella species flower thrips. Flower samples taken at least weekly are needed to understand predator-prey dynamics. Shown here is the sampling of the flowers of fruiting vegetables and companion plants to estimate the densities of individual thrips and minute pirate bug species. Representative data illustrates how the protocol is used to determine the efficacy of management tactics over time and how to evaluate the benefits of predation by minute pirate bugs. The sampling protocol is similarly adaptable to sampling thrips and minute pirate bugs in other plant species hosts.

Sampling for Estimating Frankliniella Species Flower Thrips and Orius Species Predators in Field Experiments

Presented here is a protocol to determine the number of thrips and minute pirate bug predators in crops over multiple dates in field experiments. Also illustrated is how to determine the efficacy of management tactics against thrips and evaluate the benefits of predation by minute pirate bugs.

דגימה להערכת פרנקליאלה מיני פרחים ומינים טורפים בניסויי שדה

The western flower thrips, Frankliniella occidentalis (Pergande), is a polyphagous pest that has been spread worldwide. The extensive use of insecticides ...