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>
	<li>Massa, D., Kaiser, Y. I., And&#250;jar-S&#225;nchez, D., Carmona-Alf&#233;rez, R., Mehrtens, J., Gerhards, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+geo-referenced+database+for+weed+mapping+and+analysis+of+agronomic+factors+affecting+herbicide+resistance+in+Apera+spica-venti+L.+Beauv.+(Silky+Windgrass).">Development of a geo-referenced database for weed mapping and analysis of agronomic factors affecting herbicide resistance in Apera spica-venti L. Beauv. (Silky Windgrass).</a> Agronomy. 3 (1), 13-27 (2013).</li><li>Powles, S. B., Shaner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Herbicides Resistance and World Grains. , 308 (2001).</li><li>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=Herbicide+resistance+in+Europe:+an+overview.">Herbicide resistance in Europe: an overview.</a> Proc. BCPC International Congress. , 131-138 (2005).</li><li>Jasieniuk, M., Le Corre, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+the+evolution+of+herbicide+resistance+in+weeds.">Deciphering the evolution of herbicide resistance in weeds.</a> Trends Genet. 29 (11), 649-658 (2013).</li><li>Heap, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+documentation+of+herbicide+resistance.">Identification and documentation of herbicide resistance.</a> Phytoprotection. 75 (4), 85-90 (1994).</li><li>Beckie, H. J., Heap, I. M., Smeda, R. J., Hall, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Screening+for+herbicide+resistance+in+weeds.">Screening for herbicide resistance in weeds.</a> Weed Technol. 14 (2), 428-445 (2000).</li><li>Tal, A., Kotoula-Syka, E., Rubin, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seed-bioassay+to+detect+grass+weeds+resistant+to+acetyl+coenzyme+A+carboxylase+inhibiting+herbicides.">Seed-bioassay to detect grass weeds resistant to acetyl coenzyme A carboxylase inhibiting herbicides.</a> Crop Prot. 19, 467-472 (2000).</li><li>Boutsalis, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Syngenta+Quick-Test:+a+rapid+whole-plant+test+for+herbicide+resistance.">Syngenta Quick-Test: a rapid whole-plant test for herbicide resistance.</a> Weed Technol. 15 (2), 257-263 (2001).</li><li>Menchari, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Weed+response+to+herbicides:+regional-scale+distribution+of+herbicide+resistance+alleles+in+the+grass+weed+Alopecurus+myosuroides.">Weed response to herbicides: regional-scale distribution of herbicide resistance alleles in the grass weed Alopecurus myosuroides.</a> New Phytol. 171 (4), 861-874 (2006).</li><li>Burgos, N. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review:+confirmation+of+resistance+to+herbicides+and+evaluation+of+resistance+levels.">Review: confirmation of resistance to herbicides and evaluation of resistance levels.</a> Weed Sci. 61 (1), 4-20 (2013).</li><li>Owen, M. J., Martinez, N. J., Powles, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+herbicide-resistant+Lolium+rigidum.+(annual+ryegrass)+now+dominates+across+the+Western+Australian+grain+belt.">Multiple herbicide-resistant Lolium rigidum. (annual ryegrass) now dominates across the Western Australian grain belt.</a> Weed Res. 54 (3), 314-324 (2014).</li><li>Beckie, H. J., Tardif, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herbicide+cross+resistance+in+weeds).">Herbicide cross resistance in weeds).</a> Crop Prot. 35, 15-28 (2012).</li><li>Moss, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+occurrence+of+herbicide-resistant+grass-weeds+in+the+United+Kingdom+and+a+new+system+for+designating+resistance+in+screening+assays.">The occurrence of herbicide-resistant grass-weeds in the United Kingdom and a new system for designating resistance in screening assays.</a> Proc. BCPC Weeds. , 179-184 (1999).</li><li>Baskin, C. C., Baskin, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Seeds, Ecology, Biogeography and Evolution of dormancy and Germination. , 27-42 (1998).</li><li>Sattin, M., Gasparetto, M. A., Campagna, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Situation+and+management+of+Avena+sterilis.+ssp.+ludoviciana.+and+Phalaris+paradoxa.+resistant+to+ACCase+inhibitors+in+Italy.">Situation and management of Avena sterilis. ssp. ludoviciana. and Phalaris paradoxa. resistant to ACCase inhibitors in Italy.</a> Proc. BCPC - Weeds. , 755-762 (2001).</li><li>Scarabel, L., Varotto, S., 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=A+European+biotype+of+Amaranthus+retroflexus.+cross-resistant+to+ALS+inhibitors+and+response+to+alternative+herbicides.">A European biotype of Amaranthus retroflexus. cross-resistant to ALS inhibitors and response to alternative herbicides.</a> Weed Res. 47 (6), 527-533 (2007).</li><li>Collavo, A., Panozzo, S., Lucchesi, G., Scarabel, L., 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=Characterisation+and+management+of+Phalaris+paradoxa.+resistant+to+ACCase-inhibitors.">Characterisation and management of Phalaris paradoxa. resistant to ACCase-inhibitors.</a> Crop Prot. 30 (3), 293-299 (2011).</li><li>Scarabel, L., Carraro, N., Sattin, M., Varotto, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+basis+and+genetic+characterisation+of+evolved+resistance+to+ALS-inhibitors+in+Papaver+rhoeas.">Molecular basis and genetic characterisation of evolved resistance to ALS-inhibitors in Papaver rhoeas.</a> Plant Sci. 166 (3), 703-709 (2004).</li><li>Panozzo, S., Scarabel, L., Tranel, P. 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.

عدة خطوات داخل البروتوكولات هي حاسمة لتقييم النجاح في مقاومة مبيدات الأعشاب في عدد السكان: 1) يجب أن يتم جمع البذور عندما تنضج من النباتات التي قد نجا من العلاج مبيدات الأعشاب (ق). نضج البذور على النبات الأم هو أمر حاسم لتجنب الصعوبات في إنبات البذور في وقت لاحق. 2) ينصح ?...

<html> مبيدات الأعشاب هي الأكثر استخداما على نطاق واسع التدبير مكافحة الأعشاب الضارة، وهو ما يمثل ما يصل إلى 50٪ من السوق العالمي وقاية النبات 1. هم أدوات رخيصة نسبيا، وتجنب الممارسات زراعة التربة وتستغرق وقتا طويلا كثيفة العمالة، ويؤدي في نهاية المطاف إلى إنتاج وآمنة فعالة من حيث التكلفة والمواد الغذائية مربحة 2. ومع ذلك، فإن الفيزيولوجية والوراثية كبير الحاضر تقلب في العديد من أنواع الأعشاب، جنبا إلى جنب مع الاعتماد المفرط على استخدام مبيدات الأعشاب، والنتائج في كثير من الأحيان في اختيار السكان الاعشاب مقاومة لمبيدات الأعشاب. إدخال مبيدات الأعشاب انتقائية مع هدف الأيض محددة للغاية 05/03 قد ازداد بشكل ملحوظ عدد الحالات المقاومة على مر السنين. حتى الآن، 240 أنواع الأعشاب (140 dicots و 100 ذوات الفلقة) في جميع أنحاء العالم تطورت المقاومة لمبيدات الأعشاب مواقع مختلفة عمل (SOA) 4 من. هذا هو مصدر قلق كبير لإدارة الاعشاب وأكثر من ذلك في عام لإنتاج المحاصيل المستدام. e_content &quot;&gt; الكشف المبكر عن المقاومة، بناء على اختبارات موثوقة، يقوم في كثير من الأحيان في دفيئة، هو خطوة رئيسية لإدارة الحشائش المقاومة لمبيدات الأعشاب. وقد تم تطوير مناهج مختلفة وفقا لأهداف والمستوى المطلوب من الدقة والوقت والموارد المتاحة، و كذلك أنواع الأعشاب تعتبر 12/06. ومع ذلك، عندما يطلب تأكيد حالة مقاومة الحشائش نمط حيوي جديد (أي مجموعة من الأفراد التي تشترك العديد من الخصائص الفسيولوجية، بما في ذلك القدرة على البقاء على قيد الحياة واحد أو أكثر مبيدات الأعشاب ينتمي إلى مجموعة معينة تستخدم في جرعة من شأنها أن تتحكم عادة لهم)، يحتاج إلى قوة الأحيائي مصنع كامل ليتم تنفيذها في بيئة تسيطر عليها 4، 11. A نمط حيوي نادرا ما تكون مقاومة لمبيدات الأعشاب واحدة فقط. ولذلك يتميز كل نمط حيوي من نمط المقاومة معين، أي عدد ونوع الخدمية للمبيدات الأعشاب هو مقاومة ل، ومقاومة معينةالمستوى إلى كل مبيدات الأعشاب 13. تحديد أوائل وموثوق بها لنمط الصليب أو مقاومة متعددة 5، 14 مهم لإدارة المقاومة المجال. ومن الجدير بالذكر أن مقاومة مبيدات الأعشاب له علاقة مع التسامح الطبيعي شيئا أن بعض أنواع الأعشاب المعرض نحو بعض مبيدات الأعشاب، مثل الأنواع dicot مقابل ACCase المثبط للمبيدات الأعشاب، والأنواع monocot مقابل 2،4-D، كنباث الحقول مقابل الغليفوسات. تقدم هذه الورقة نهجا قويا لاختبار biotypes المقاومة لمبيدات الأعشاب المفترض عينات في المجالات التي تم الإبلاغ عن ضعف الرقابة من قبل مبيدات الأعشاب (ق). وتعرض المتغيرات ذات الصلة البروتوكولات القياسية فيما يتعلق الحشائش المعنية. ترتبط المزايا أكثر من التقنيات البديلة / البروتوكولات على أساس إما اختبارات بيولوجية مصنع كامل باستخدام واحد فقط جرعة مبيدات الأعشاب 15، أو بذور علاج في أطباق بتري 8 إلى reliab العاليility وإمكانية استنتاج مستوى المقاومة بسبب إدراج جرعتين مبيدات الأعشاب في التجارب. ومع ذلك، لاختبار مقاومة الروتيني، نفس الأساليب يمكن تطبيقها في جرعة مبيد واحد فقط، مما يقلل من التكاليف. فضلا عن السماح تأكيدا للمكانة المقاومة، فإن المعلومات التي تم الحصول عليها يمكن استخدامها على حد سواء الاستفادة المثلى من خطوات البحث التالية و / أو وضع استراتيجيات الإدارة السليمة للمقاومة.

<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.

لتقييم حالة مقاومة للسكان مقاومة المفترض، أنه أمر أساسي لتشمل الاختيار عرضة في الفحص من أجل التأكد من فعالية مبيدات الأعشاب. نتائج اختبار الفحص التي أجريت على P. السكان rhoeas، والأعشاب الضارة التي تغزو حقول القمح، والتي أعلن عنها في الشكل 2، حيث تم عر...

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.

بروتوكولات قوية مبيدات الأعشاب اختبار المقاومة بأنواع الأعشاب مختلفة

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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14.5K 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

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

تقنية لشاشة الزان الأمريكية لمقاومة الحشرات والزان مقياس (<em&gt; المستخفية fagisuga</em&gt; ليند.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

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.

تقنية التخصيب الأمثل للعزل<em&gt; الفصلاء</em&gt; أنواع الجراثيم من عزلات عينة التربة

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.

قياس معدلات المبيدات الأيض في Dicot الأعشاب مع الفحص رفعه ليف

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.

عند جهد اختبار التآكل

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.

بروتوكولات لقياس متبقيات المبيدات القابلة للتحويل في أنظمة المروج

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.

تشتت من المواد النانوية في الوسط المائي: نحو التحسين البروتوكول

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 والتصوير الطيفي الجذر فينوتيبينج النباتية والبحوث الفسيولوجية: الإعداد التجريبية والتصوير البروتوكولات

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.

بروتوكولات النظام الإيكولوجي تلفيق (اكوفاب) لتشييد مختبر النظم الإيكولوجية تهدف إلى دراسة التفاعلات بين الميكروبات والنبات

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

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol for performing in situ juvenile exposure tests within oligotrophic river catchments over one-month and three-month periods. Two methods (in both modifications) are presented to evaluate the juvenile growth and survival rate. The methods and modifications differ in value for the locality bioindication and each has its benefits as well as limitations. The sandy cage method works with a large set of individuals, but only some of the individuals are measured and the results are evaluated in bulk. In the mesh cage method, the individuals are kept and measured separately, but a low individual number is evaluated. The open water exposure modification is relatively easy to apply; it shows the juvenile growth potential of sites and can also be effective for water toxicity testing. The within-bed exposure modification needs a high workload but is closer to the conditions of a natural juvenile environment and it is better for reporting the real suitability of localities. On the other hand, more replications are needed in this modification due to its high-hyporheic environment variability.

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

In situ bioindications enable determination of the suitability of an environment for endangered mussel species. We describe two methods based on the juvenile exposure of freshwater pearl mussels in cages to oligotrophic river habitats. Both methods are implemented in variants for open water and hyporheic water environments.

بيوينديكيشن اختبار لمدى ملاءمة البيئة تيار للشباب المياه العذبة بيرل بلح البحر باستخدام طرق التعرض في الموقع

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

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.

تكوين وتحليل التوزيع من بيوايروسولس تحت ظروف بيئية مختلفة

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