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.

1. Tohum Örnekleme ve Depolama <ol><li> Olumsuz iklim koşulları ya da düşük kaliteli herbisit tedavileri nedeniyle değil, yani haksız kötü herbisit performansı için ekili alanlar, monitör. </li><li> Her seferinde, bir türden gelen bir tohum numunesi toplamak ve özgün bir kod atar. Olgun tohumlar genellikle herbisit tedavi (ler) hayatta olan bitkilerden mahsul hasat öncesi toplanır. Tohumlar ne zaman anne bitki tarafından olgun döken eğer zamanında monitör gözlemlemek. </li><li> Türler, toplama tarihi, GPS koordinatları, belediyenin, çiftçinin adı, alan büyüklüğü, istila düzeyi, kırpma, herbisit (ler) sezonunda kullanılan ve alanın tarihsel kayıtların atanan benzersiz bir kod adını gösteren her bir numune için formu doldurunuz . </li><li> Alan istilası temsilcisi olan en az 30 rastgele seçilmiş bitkilerden tohum toplayın. Tohum örneği, en az 5.000 olgun tohumları içerdiğinden emin olun. Zorunlu bir döllenmedir yabani ot türleri için(Örneğin, lolium spp., Veya Amaranthus spp.), 5000 11 civarında toplam tohum sayısı, tutmak, 10-15 bitkilerin sayısını azaltır. </li><li> Yabancı otların yamalar (hektar daha fazla) geniş alanlar üzerinde dağınık sanki farklı herbisit dirençli biyotipleri seçilen alanı Alt-örnek. </li><li> Atanan benzersiz kod ile etiketlenmiş damlar kağıt torbalarda saklayın tohumları. </li><li> Nemin buharlaşmasını ancak ikincil uyuşukluk uyarılmasını önlemek için yüksek sıcaklık (yani, güneş altında bir arabada bırakarak önlemek) veya aşırı sıcaklık dalgalanmaları tohum maruz bırakmayın izin verin. </li><li> (Saman, de-gövde tohum, vb kaldırmak) ve kuru bir odada ortam sıcaklığında saklayın temizleyin. İlk direnç testler sonrasında, vakumlu plastik torbalarda, tercihen 4 ° C&#39;de karanlık bir odada uzun süre tohumlarını saklayın. Bu şekilde tohumları önemli ölçüde daha uzun bir süre boyunca, yaşama yetenekleri korur. </li></ol> NOT: Tohum dinlenmesi adapte ve tarımsal ekosistemlerde kalıcı yabani otlar sağlayan esnek ve verimli bir mekanizma sağlar. Tohum çimlenme dinlenmesi kırmak ve izin vermek için, farklı protokoller örneğin zararlı bitki türleri, uyuşukluk 16 tipine bağlı olarak kullanılabilir olması gerekir. Dormansi kaldırmak için üç ana yöntem vardır: <ol><li> Vernalizasyon NOT:. Örn Amaranthus retroflexus, Chenopodium albüm, Lolium spp Avena fatua, Polygonum persicaria&#39;ya: eşzamanlı çimlenmesini ve fide çıkmasını, bir hafta, birkaç gün arasında değişen tohum vernalizasyon bir süre elde etmek için birçok türe fizyolojik dormansi kaldırmak için gereklidir , Phalaris 17-19 paradoxa. Kadar 15 gün daha uzun bir süre Schoenoplectus mukronatus için en fazla 30 gün Papaver rhoeas, Cyperus difformis ve Ammania coccinea için gereklidir ve <sup&gt; 20. <ol><li> Plastik tabaklar içinde bazı deiyonize su koyun. Herhangi bir aşırı kaldırmak, filtre kağıdı iki kat kesin ve suda bekletin. Kağıt üzerinde hava kurutulmuş tohumları yerleştirin. Gereken zaman süresi için, 4 ° C&#39;de bir buzdolabı plastik tabaklar aktarın. </li></ol></li><li> Kazıma NOT: Bazı yabancı ot türlerinin nedeniyle mekanik dormansi, yani tohum kabuğu özelliklerine diğerlerinden daha çimlenme daha inatçı, ve 21 çimlenmeye sülfürik asit kullanılarak kimyasal kazıma kullanımını gerektirmektedir. <ol><li> Konsantre sülfürik asit (% 95-98) ile bir çanak hazırlayın. Su dolu bir beher hazırlayın. Dokuma olmayan kumaşın bir zarf içinde tohumlar konulur. </li><li> Konsantre sülfürik asit içinde, sırası ile, 20 dakika ya da 5 dakika boyunca, örneğin, echinochloa spp., Veya Sorghum halepense tohumları bekletin. </li><li> Cımbız, bir çifti kullanılarak beher dışına zarfı alın ve su dolu behere koyun. Açzarf, küçük bir kevgir içinde tohum koymak ve akan su altında iyice yıkayın. </li></ol> Varyant: A. plantago-aquatica farklı bir protokol ihtiyacı 22, 23 <ol><li> Kloroform içinde 2 dakika için tohum bekletin. Deiyonize su ile durulayın ve tohumların emici kağıt ile kurulayın. 5 dakika için% 80 sülfürik asit içinde tohum Dunk. </li><li> Küçük bir kevgir içinde tohum koyun ve akan su altında iyice yıkayın. </li></ol></li><li> Hasat sonrası tohum olgunlaşma NOT: Diğer yabancı ot türlerinin tohumları bakılmaksızın dinlenmesi kırmak için kullanılan yöntem, olgunluk sonra bir kaç ay hiç çimlenme yok. <ol><li> RT ve düşük nem en az 3-4 aylık bir süre için tohum saklayın ve daha sonra dinlenmesi kırma (örneğin, Oryza sativa var. Sylvatica veya P. rhoeas) için yukarıdaki protokolleri uygulayın. </li></ol></li></ol> 3. Tohum Çimlenme <ol><li> Talep tohumları plas çimlenmiş edilmesi% 0.1 potasyum nitrat (KNO 3) ile% 0.6 (kütle / hacim) agar içeren tlc yemekler eklendi: <ol><li> Iyonu giderilmiş su kullanarak,% 0.6 +% 0.1 KNO 3&#39;te agar ihtiva eden bir çözelti hazırlayın. Bir mikrodalga fırında agar çözülür. </li><li> Plastik tabaklar içine agar çözüm dökün. Substrat soğutun ve daha sonra tohumlarda koydu. </li></ol> Varyasyon: Toprak da plastik tabaklar bir alt-tabaka olarak kullanılabilir. Bu uygulama Echinochloa spp için özellikle verimlidir., S. halepense ve Lolium spp. </li><li> Her ot türlerinin optimum koşullara bağlı olarak hafif ve sıcaklık koşulları ile ilgili bir hafta boyunca bir çimlenme dolabında plastik yemekleri yerleştirin. En kış türleri için, sıcaklık aralığı 15/25 ° C, gece / gündüz ve 15-30 ľmol m -2 sn -1 bir Fotosentez Foton Akı Yoğunluğu (PPFD) sağlayan neon tüpleri ile 12 saat fotoperiyot olduğunu. Birçok yaz türleri için, sıcaklık aralığı 15/30 ° C gece / gün&#39;dür. Varyant: SBöyle S. olarak ome türler, halepense, bir ısıl işlem gerekir. S. nedenle kazıma sonra, tohumlar normal şartlarda daha sonra üç gün çimlenme odasında üç gün için 24 ° C &#39;de 45 ° C ve 20 saat sonra, 4 saat döngüleri ve: halepense aşağıdaki koşullara tabi tutulurlar. </li></ol> 4. Fide Fide ve Büyüme <ol><li> Nakli standart çömlekçilik karışımı ile dolu plastik tepsiler (325 x 265 x 95 mm) içine on beş-yirmi fidanları (% 60 siltli tınlı toprak,% 15 kum,% 15 ve% 10 agriperlite turba - hacimce). Not: Fide yerine doğrudan ekim, herbisit uygulamasının performansını optimize etmek için önemli bir ön-koşul olan aynı büyüme aşamasında bitkilerin üniform bir standı elde edilmesini sağlar. </li><li> Numarası ve ilerici tepsi numarasını çoğaltmak, nüfus kodu, herbisit test edilen &lt;: Eşsiz tanımlanması için tüm bilgiler de dahil olmak üzere bir barkod her tepsiyi tanımlayın./ Li&gt; <li> Ya da tarla kapasitesi yakınındaki alt tabakayı korumak için gerektiği gibi bir ısıtmalı sera ve su bitkileri koyun tepsileri. Not: büyüme sıcaklığı yabani ot türlerine bağlı olarak değişir. Genellikle testler sonbahar / kış / bahar aylarında yapılır, bu nedenle ışık yaklaşık 150 umol m -2 sn PPFD sağlayan 400 W metal halide lambalar, -1 ve 12 saat fotoperiyodun 24 19. Yaz ot türleri ile kullanılarak güçleniyor C 4 fotosentez döngüsü genellikle daha yüksek ışık yoğunluğu gerektirir ve bu nedenle testlerin geç ilkbahar-yaz yapılan veya takviye ışık yoğunluğu yaklaşık 400 umol m -2 s -1 14 saat fotoperyod ile vardır. </li><li> Çeltik, örneğin, A. infesting bazı ot türleri için farklı bir protokol kullanın plantago-aquatica, S. mukronatus ve C 22 de tarif edildiği gibi difformis. <ol><li> 24 yuvarlak hücreler (55 mm çapında, 64 mm derinlik) fil ile polistiren tepsilerine fidan nakli% 60 silt killi toprak,% 30 kum ve (hacimce)% 10 turba ile olmuştur. </li><li> 12 cm tepsileri derin plastik kaplar su ile doldurulmuş ve onları (Şekil 1) yüzen önlemek için vidalı paslanmaz çelik çubuklar aşağı battened ayarlayın. </li><li> Toprak yüzeyinin seviyesinin altında 1-2 cm kaplarda su seviyesini korumak ve yosun çoğalmasını önlemek için su (10-12 L içeren) her bir kaba bakır sülfat ve 1.5 g ekleyin. </li></ol></li></ol> 5. Herbisit Tedaviler <ol><li> Acil durum öncesi herbisidler olan tedaviler: <ol><li> Çimlenme kabininde yaklaşık üç gün bölüm 3&#39;te tarif edildiği gibi sonra, yukarıda tarif edilen alt tabaka içeren plastik tepsilere çimlenen tohumların ekimi ve toprak tabakası (yaklaşık 1 cm) ile kaplayın. Bu fideler herbisit etkisi ziyade aşırı mezar derinliği nedeniyle ortaya olmaz sağlamak için kritik bir adımdır. </li><li> Alan dikişleri alt tabakanın atınsu dolu tabaklar üzerinde, altta bir kaç delik var tepsiler, yerleştirerek şehir. </li><li> Bir gün dikimden sonra, filizlenme öncesi herbisit 25 tepsileri davranın. </li><li> Daire dan kapilarite tarafından yukarıdan ve aşağıdan hem gerektiği gibi su ekleyerek tabakayı veya yakın alan kapasitesi tutun. Bu prosedür iyi bir tedavi etkinliği için (yani, çimlenen tohumlar nerede) uygun derinlikte herbisit kalıcılığını yanadır. </li></ol></li><li> Sonrası ortaya Herbisitlerle Tedaviler: <ol><li> 2-3 yaprak aşamasına ulaştığınızda bitkiler Sprey (yani büyüme evresi Genişletilmiş BBCH büyüme ölçek 26 12-13). </li><li> Tedaviden sonra günden itibaren, yabani ot türlerinin su gereksinimleri ve echinochloa spp sezonu (örneğin, uygun sulama sistemini ayarlayın. O 9&#39;a 09:00 dan düzenli aralıklarla, 3 dakika boyunca günde 4 kez su sunar pm). Su dis olduğunuotomatik yağmurlama sulama sistemi kullanılarak tributed. Varyant: glifosat bitki sahne BBCH 14-21 uygulanır. </li></ol></li><li> Herbisit hazırlama ve dağıtımı. Not: (ön ve son-ortaya çıkma) Bütün herbisitler, iki doz tavsiye edilen yüzey aktif maddeler ile, ticari formülasyonlar olarak uygulanır önerilen alan dozu (1 x) ve üç kez (3x) olduğu. <ol><li> Gerekirse, etiket talimatlarına göre toplu olarak surfaktan çözüm hazırlamak; nihai konsantrasyon genellikle nihai hacminin bir yüzdesi (örneğin,% 0.3) halinde ifade edilir ya da hacim olarak birim alan (örneğin, 1 L ha-1) ortalama dağıtılacak. </li><li> Aktif terkip maddesinin doğru konsantrasyon tutmak için herbisit (çözünen) çözeltisi için çözücü olarak yüzey aktif madde solüsyonu kullanın. İlk en yoğun herbisit çözeltisi hazırlayın (3x). Ticari ürünün miktarı yüzey aktif madde çözeltisi içinde çözüldü (ya da, eğer, iyonu giderilmiş su içinde olmak hesaplayınyüzey aktif madde, aşağıdaki denklem kullanılarak) gerekli değildir: Doz ot = [(Doz alan x Doz max) x V fin] / V del Burada: Doz ot = herbisit dozu (mi), Doz alanı = herbisit alan dozu (ha mi-1), Doz maks = maksimum dozun, V kanatçık = çözeltisi (L) son hacmi, V del tarafından sağlanan hacim = tezgah püskürtücü (L ha -1). </li><li> Daha az konsantre bir (1x) maddesinin hazırlanması için: (1, hacim / hacim olarak 2) herbisit solüsyonu 3x seyreltilir. Bu prosedür tartı veya herbisit pipetleme yaparken hata yapma olasılığını azaltır. Birim, birim alan (L ha-1) ortalama dağıtılacak şekilde Herbisit solüsyonu konsantrasyonu olarak ifade edilir. </li><li> Alt herbisit dozunun (1x) ile muamele dizisini başlatın. Bu şekilde, aynı herbisit ile iki tedavi arasında püskürtme kabini yıkamak için bir ihtiyaç vardır. </li><li> Herbisit Solutio dağıtınn üç düz fan (genişletilmiş aralık) hidrolik nozulları ile donatılmış bir patlama ile 215 kPa basınçta 300 L ha -1 (±% 1), teslim hassas tezgah püskürtücü, ve 0.75 m sn -1 hıza kullanarak . </li><li> Herbisit ağartıcının% 1 ile değiştirildiğinde iki kez, püskürtme kabini yıkayın (h / h) ve daha sonra durulanır. Varyasyon: glifosat, 200 L ha -1 27 bir püskürtme hacmi ile tatbik edilir. NOT: Özellikle dikkat yüksek biyolojik herbisitler, bu tür sülfonilüreler sülfometuron veya flazasulfuron, kullanıldığı zaman ödenmesi gerekir. Bu son durumda, amonyak ile, bir bir ağartma çözeltisi ile yıkama ve bir başkası da (2.5% h / h) su ile dikkatli bir şekilde durulanmıştır. </li></ol></li></ol> 6. Toplama ve Verilerin Analizi <ol><li> Otomatik olarak her tepsiyi tanımlayan bir barkod okuyucu sayesinde, tedavi yanı sıra görsel Tahmini Biyokütle (VEB) hayatta bitkilerin sayısını kaydedin. Bitkiler eşek vardırne olursa olsun renk ya da diğer görünüm aktif büyüme gösteriyorsa ölü olarak Essed. <ol><li> Test herbisit bağlı olarak üç ya da dört hafta tedavi (WAT) sonra değerlendirme yapmak (örneğin, ACCase inhibitörleri üç WAT ve ALS inhibitörleri ya da glifosata dört WAT). </li><li> Yani duyarlı nüfus (S edin) bütün deneylerde, asla veya nadiren herbisit ile tedavi olan bir site toplanan bir nüfusa dahil edilerek genel tedavi etkinliğini değerlendirmek. </li><li> Işlenmiş bitkilerin sayısının yüzdesi olarak Express bitki hayatta kalma, (iki çoğaltır ortalama değer) sadece herbisit tedavi öncesi sayılır ve ortalama değer başına standart hata (SE) hesaplayın. </li><li> Tedavi ve aynı nüfus 25, 28 check-tedavi edilmezse arasındaki VEB bitki biyokütle bir görsel karşılaştırma yoluyla elde edilir. Bir puan, 0 (değil tedavi edilen kontrol ile karşılaştırıldığında) herbisit etkilenmeyen bitkiler için 10 arasında değişen zamanbitkiler açıkça ölü, her tedavi tepsiye verilir. </li></ol></li><li> S kurtulanlar daha fazla herbisit dozu 1x R 20% 5 ila% arasında değişmekteydi zaman bitkilerin% 5&#39;inden az herbisit dozu 1x, SR hayatta: İki herbisit dozu ile tedavi elde edilen sonuçlara dayalı olarak dört kategoride ascribe popülasyonları Hayatta kalanlar herbisit dozu 1x de% 20&#39;den fazla ve herbisit dozu 3x 17 de% 10&#39;dan fazla olduğunda bitkilerin% 20 herbisit dozu 1x ve RR atlattı. </li></ol>

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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.

1) tohumları, olgun, herbisite muamele (ler) hayatta kalan bitkiler temin edilmelidir: protokoller dahilinde birçok adım bir popülasyonda herbisit direnci başarılı bir şekilde değerlendirilmesi için kritik öneme sahiptir. Anne bitki tohumlarının olgunlaşması daha sonra tohum çimlenmesi zorluklar önlemek için çok önemlidir; 2) tohumların çimlenmesini uygun depolama önleyecek kalıp çoğalmasını önlemek için tavsiye edilir; Herbisit paketinin etiketinde bildirilen 3) fidanları, sağ büyüme a...

<html> Herbisitler küresel bitki koruma piyasasının 1 kadar% 50&#39;sini, en yaygın kullanılan yabancı ot kontrolü önlemi vardır. Onlar emek yoğun ve zaman alıcı bir toprak işleme uygulamaları önlemek ve sonuçta maliyet-etkin, güvenli ve kârlı gıda üretimi 2 sonuçlanır, nispeten ucuz araçlardır. Ancak, birlikte herbisit kullanımı ile ilgili bir aşırı güven ile birçok yabani ot türlerinde büyük fenolojik ve genetik değişkenlik mevcut, sık sık herbisit dirençli yabani ot popülasyonlarının seçiminde sonuçlanır. çok özel bir metabolik hedefle seçici herbisit tanıtımı 3-5 yılda direnç vakalarının sayısını büyük oranda arttırmıştır. Bugüne kadar dünya çapında 240 yabancı ot türlerinin (140 dikotlar ve 100 monocots) Eylem (SoA) 4 farklı herbisit Siteleri direnç geliştirmişlerdir. Bu sürdürülebilir bitkisel üretim için önemli bir ot yönetimi için endişe ve genel olarak daha fazladır. Sık sık bir serada gerçekleştirilen güvenilir testler, dayalı e_content &quot;&gt; direnişin Erken teşhis, herbisit dirençli yabani otları yönetmek için önemli bir adımdır. Farklı yaklaşımlar olarak, amaçları, doğruluk, zaman ve mevcut kaynakların gereken seviyeye göre geliştirilmiştir de 6-12 olarak yabani ot türü gibi. Ancak, yeni bir yabancı ot biyotip direnç durumunun onayı (gerekli olduğu zaman, yani, bir ait olan bir ya da daha fazla herbisit hayatta yeteneği dahil olmak üzere birçok fizyolojik özellikleri paylaşan bireylerin bir grubu, normalde bunları kontrol edecek bir dozda kullanılan özel grup), sağlam bir bütün bitki biyo-deney, kontrollü bir ortamda 4, 11 yapılmaktadır gerekmektedir. Bir biyotip sadece bir herbisit nadiren dayanıklıdır. Her bir biyotip bu nedenle dayanıklıdır herbisitlerin belirli bir direnç desen, SOA, yani sayısına ve türüne göre, özelliği, ve belirli bir direnci olanHer bir bitki öldürücü 13 düzeyi. haç veya birden fazla direnç 5 Desenin erken ve güvenilir bir tespiti, 14 alan direnç yönetimi için çok önemlidir. O herbisit direnci doğal tolerans ile ilgisi vardır kayda değer bazı herbisit, örneğin, ACCase önleyici herbisitler, monokot türlerinin vs 2,4-D vs dikot türler, equisetum arvense vs yönünde bazı ot türlerinin sergi glifosat. Bu makale herbisit (ler) tarafından yoksul kontrolü olduğu bildirildi alanlarda örneklenmiş varsayılan herbisit dirençli biyotiplerin test etmek için sağlam bir yaklaşım sunuyor. Katılan yabani ot türleri ile ilgili olarak, standart protokollere ilgili varyantlar sunulmaktadır. Petri 8 tek herbisit dozu 15 ya da tedavi tohum kullanan ya bütün bitki biyoanalizlerin dayalı alternatif teknikler / protokoller üzerinden avantajları yüksek reliab ilgiliility nedeniyle deneylerde iki herbisit dozu dahil direnç seviyesini çıkarım olasılığı. Ancak rutin direnci testi için, aynı yöntemler, böylece maliyetlerin azaltılması, tek bir herbisit dozunda uygulanabilir. Yanı sıra direnç durumunun onayı izin olarak, elde edilen bilgiler hem aşağıdaki araştırma adımlarını optimize ve / veya ses direnç yönetimi stratejileri oluşturulması için kullanılabilir.

<table><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 17 g m&lt;sup&gt;-2&lt;/sup&gt;</td></tr><tr><td>Chloroform &gt;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 &gt;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>537 x 328 x 72 mm, 24 round cells (6x4)</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>Germination cabinet</td><td>KW</td><td>W87R</td><td></td></tr><tr><td>Nozzles</td><td>Teejet</td><td>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 colspan="4">&lt;strong&gt;Herbicides included in the results:&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Commercial product&lt;/strong&gt;</td><td>&lt;strong&gt;Active ingredient&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>Altorex</td><td>imazamox</td><td>BASF</td><td></td></tr><tr><td>Azimut</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</td><td>tribenuron-methyl</td><td>Dupont</td><td></td></tr><tr><td>Gulliver</td><td>azimsulfuron</td><td>Dupont</td><td></td></tr><tr><td>Hussar WG</td><td>iodosulfuron</td><td>Bayer Crop Science</td><td></td></tr><tr><td>Nominee</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</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.

Bir dayanıklı varsayılan nüfusun direnci durumunu değerlendirmek için, herbisit etkinliğini doğrulamak için tahlilde duyarlı bir kontrol dahil etmek esastır. P. üzerinde yapılan bir tarama testinin sonuçları rhoeas popülasyonları, buğday alanları infesting bir ot, bir duyarlı kontrol (09-36) ve şüpheli dirençli bir (10-91) dört sonrası ortaya herbisit etkinliği sunulmaktadır Şekil 2&#39;de, raporlanır. Nüfus 09-36 tamamen tek bir bitki, florasulam ve tribe...

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.

Farklı Ot Türlerinin Sağlam Herbisit Direnci Testi için Protokoller

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

protocols-for-robust-herbicide-resistance-testing-different-weed

Universidad San Sebastian

Research

JoVE Journal

Environment

14.5K Görüntüleme.  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

Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions

Gene deletion plays an important role in the analysis of gene function. One of the most efficient methods to disrupt genes in a targeted manner is the replacement of the entire gene with a selectable marker via homologous recombination. During homologous recombination, exchange of DNA takes place between sequences with high similarity. Therefore, linear genomic sequences flanking a target gene can be used to specifically direct a selectable marker to the desired integration site. Blunt ends of the deletion construct activate the cell&#39;s DNA repair systems and thereby promote integration of the construct either via homologous recombination or by non-homologous-end-joining. In organisms with efficient homologous recombination, the rate of successful gene deletion can reach more than 50% making this strategy a valuable gene disruption system. The smut fungus Ustilago maydis is a eukaryotic model microorganism showing such efficient homologous recombination. Out of its about 6,900 genes, many have been functionally characterized with the help of deletion mutants, and repeated failure of gene replacement attempts points at essential function of the gene. Subsequent characterization of the gene function by tagging with fluorescent markers or mutations of predicted domains also relies on DNA exchange via homologous recombination. Here, we present the U. maydis strain generation strategy in detail using the simplest example, the gene deletion.

Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions

We describe a robust gene replacement strategy to genetically manipulate the smut fungus Ustilago maydis. This protocol explains how to generate deletion mutants to investigate infection phenotypes. It can be extended to modify genes in any desired way, e.g., by adding a sequence encoding a fluorescent protein tag.

Bitki patojeni olarak genetik manipülasyonu<em&gt; Ustilago maydis</em&gt; Mantar Biyoloji ve Bitki Mikrop Etkileşimleri Eğitim için

Gene deletion plays an important role in the analysis of gene function. One of the most efficient methods to disrupt genes in a targeted manner is the ...

Genetik

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.

Bir kesilmiş yaprak Testi ile Dikot Weeds Herbisit Metabolizma düşük Ölçme

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

Çevre

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.

Çim Bitkileri Sistemlerinde Transferli Pestisit Kalıntı miktarının için Protokoller

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

Screening Cotton Genotypes for Reniform Nematode Resistance

A rapid non-destructive reniform nematode (Rotylenchulus reniformis) screening protocol is needed for the development of resistant cotton (Gossypium hirsutum) varieties to improve nematode management. Most protocols involve extracting vermiform nematodes or eggs from the cotton root system or potting soil to determine population density or reproduction rate. These approaches are generally time-consuming with a small number of genotypes evaluated. An alternative approach is described here in which the root system is visually examined for nematode infection. The protocol involves inoculating cotton seedling 7 days after planting with vermiform nematodes and determining the number of females attached to the root system 28 days after inoculation. Data are expressed as the number of females per gram of fresh root weight to adjust for variation in root growth. The protocol provides an excellent method for evaluating host-plant resistance associated with the ability of the nematode to establish an infection site; however, resistance that hinders nematode reproduction is not assessed. As with other screening protocols, variation is commonly observed in nematode infection among individual genotypes within and between experiments. Data are presented to illustrate the range of variation observed using the protocol. To adjust for this variation, control genotypes are included in experiments. Nonetheless, the protocol provides a simple and rapid method to evaluate host-plant resistance. The protocol has been successfully used to identify resistant accessions from the G. arboreum germplasm collection and evaluate segregating populations of more than 300 individuals to determine the genetics of resistance. A vegetative propagation method for recovering plants for resistance breeding was also developed. After removal of the root system for nematode evaluation, the vegetative shoot is replanted to allow the development of a new root system. More than 95% of the shoots typically develop a new root system with plants reaching maturity.

Here, a protocol is presented for the rapid non-destructive screening of cotton genotypes for reniform nematode resistance. The protocol involves visually examining the roots of nematode-infected cotton seedlings to determine infection response. The vegetative shoot from each plant is then propagated to recover plants for seed production.

Reniform Nematode direnci için pamuk Genotürleri eleme

A rapid non-destructive reniform nematode (Rotylenchulus reniformis) screening protocol is needed for the development of resistant cotton (Gossypium ...

Quantification of the Potential Impact of Glyphosate-Based Products on Microbiomes

Glyphosate-based products (GBP) are the most common broad-spectrum herbicides worldwide. The target of glyphosate is the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in the shikimate pathway, which is virtually universal in plants. The inhibition of the enzyme stops the production of three essential amino acids: phenylalanine, tyrosine, and tryptophan. EPSPS is also present in fungi and prokaryotes, such as archaea and bacteria; thus, the use of GBP may have an impact on the microbiome composition of soils, plants, herbivores, and secondary consumers. This article aims to present general guidelines to assess the effect of GBP on microbiomes from field experiments to bioinformatics analyses and provide a few testable hypotheses. Two field experiments are presented to test the GBP on non-target organisms. First, plant-associated microbes from 10 replicated control and GBP treatment plots simulating no-till cropping are sampled and analyzed. In the second experiment, samples from experimental plots fertilized by either poultry manure containing glyphosate residues or non-treated control manure were obtained. Bioinformatics analysis of EPSPS protein sequences is utilized to determine the potential sensitivity of microbes to glyphosate. The first step in estimating the effect of GBP on microbiomes is to determine their potential sensitivity to the target enzyme (EPSPS). Microbial sequences can be obtained either from public repositories or by means of PCR amplification. However, in the majority of field studies, microbiome composition has been determined based on universal DNA markers such as the 16S rRNA and the internal transcribed spacer (ITS). In these cases, sensitivity to glyphosate can only be estimated through a probabilistic analysis of EPSPS sequences using closely related species. The quantification of the potential sensitivity of organisms to glyphosate, based on the EPSPS enzyme, provides a robust approach for further experiments to study target and non-target resistant mechanisms.

Glyphosate-based products (GBP) are the most common broad-spectrum herbicides worldwide. In this article, we introduce general guidelines to quantify the effect of GBP on microbiomes, from field experiments to bioinformatics analyses.

Glifosat Bazlı Ürünlerin Mikrobiyomlar Üzerindeki Potansiyel Etkisinin Ölçülmesi

Glyphosate-based products (GBP) are the most common broad-spectrum herbicides worldwide. The target of glyphosate is the enzyme ...

Bioassays for Monitoring Insecticide Resistance

Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products are labeled for an individual pest within a particular crop system, chemical control options are limited. Therefore, the same product(s) are used repeatedly and continual selection pressure is placed on the target pest. There are both financial and environmental costs associated with the development of resistant populations. The cost of pesticide resistance has been estimated at approximately $ 1.5 billion annually in the United States. This paper will describe protocols, currently used to monitor arthropod (specifically insects) populations for the development of resistance. The adult vial test is used to measure the toxicity to contact insecticides and a modification of this test is used for plant-systemic insecticides. In these bioassays, insects are exposed to technical grade insecticide and responses (mortality) recorded at a specific post-exposure interval. The mortality data are subjected to Log Dose probit analysis to generate estimates of a lethal concentration that provides mortality to 50% (LC50) of the target populations and a series of confidence limits (CL's) as estimates of data variability. When these data are collected for a range of insecticide-susceptible populations, the LC50 can be used as baseline data for future monitoring purposes. After populations have been exposed to products, the results can be compared to a previously determined LC50 using the same methodology.

This manuscript demonstrates and discusses techniques used to survey pesticide susceptibility and detect resistance to contact and systemic pesticides in arthropod pests.

İnsektisit Direnci İzleme için biyoanalizlere

Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products ...

Biyoloji

Implementation of In Vitro Drug Resistance Assays: Maximizing the Potential for Uncovering Clinically Relevant Resistance Mechanisms

Although targeted therapies are initially effective, resistance inevitably emerges. Several methods, such as genetic analysis of resistant clinical specimens, have been applied to uncover these resistance mechanisms to facilitate follow-up care. Although these approaches have led to clinically relevant discoveries, difficulties in attaining the relevant patient material or in deconvoluting the genomic data collected from these specimens have severely hampered the path towards a cure. To this end, we here describe a tool for expeditious discovery that may guide improvement in first-line therapies and alternative clinical management strategies. By coupling preclinical in vitro or in vivo drug selection with next-generation sequencing, it is possible to identify genomic structural variations and/or gene expression alterations that may serve as functional drivers of resistance. This approach facilitates the spontaneous emergence of alterations, enhancing the probability that these mechanisms may be observed in the patients. In this protocol we provide guidelines to maximize the potential for uncovering single nucleotide variants that drive resistance using adherent lines.

Implementation of In Vitro Drug Resistance Assays: Maximizing the Potential for Uncovering Clinically Relevant Resistance Mechanisms

Drug resistance to targeted therapeutics is widespread and the need to identify mechanisms of resistance--prior to or following clinical onset--is critical for guiding alternative clinical management strategies. Here, we present a protocol to couple derivation of drug-resistant lines in vitro with sequencing to expedite discovery of these mechanisms.

Uygulanması<em&gt; İn Vitro</em&gt; İlaç Direnci Tahliller: Klinik İlgili Direnç Mekanizmaları ortaya çıkararak yönelik Potansiyeli maksimize

Although targeted therapies are initially effective, resistance inevitably emerges. Several methods, such as genetic analysis of resistant clinical ...

Tıp

Characterizing Herbivore Resistance Mechanisms: Spittlebugs on Brachiaria spp. as an Example

Plants can resist herbivore damage through three broad mechanisms: antixenosis, antibiosis and tolerance1. Antixenosis is the degree to which the plant is avoided when the herbivore is able to select other plants2. Antibiosis is the degree to which the plant affects the fitness of the herbivore feeding on it1.Tolerance is the degree to which the plant can withstand or repair damage caused by the herbivore, without compromising the herbivore's growth and reproduction1. The durability of herbivore resistance in an agricultural setting depends to a great extent on the resistance mechanism favored during crop breeding efforts3.
We demonstrate a no-choice experiment designed to estimate the relative contributions of antibiosis and tolerance to spittlebug resistance in Brachiaria spp. Several species of African grasses of the genus Brachiaria are valuable forage and pasture plants in the Neotropics, but they can be severely challenged by several native species of spittlebugs (Hemiptera: Cercopidae)4.To assess their resistance to spittlebugs, plants are vegetatively-propagated by stem cuttings and allowed to grow for approximately one month, allowing the growth of superficial roots on which spittlebugs can feed. At that point, each test plant is individually challenged with six spittlebug eggs near hatching. Infestations are allowed to progress for one month before evaluating plant damage and insect survival. Scoring plant damage provides an estimate of tolerance while scoring insect survival provides an estimate of antibiosis. This protocol has facilitated our plant breeding objective to enhance spittlebug resistance in commercial brachiariagrases5.

Characterizing Herbivore Resistance Mechanisms: Spittlebugs on Brachiaria spp. as an Example

This video explains mechanisms of host plant resistance to herbivory and demonstrates a no-choice test that estimates the relative contributions of antibiosis and tolerance to spittlebug resistance in Brachiaria spp.

Karakterize Otobur Direnç Mekanizmaları: Spittlebugs Brachiaria Spp. Bir Örnek

Plants can resist herbivore damage through three broad mechanisms: antixenosis, antibiosis and tolerance1. Antixenosis is the degree to which the plant is ...

A PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica

Wild-type I. cylindrica (cogongrass) is one of the top ten worst invasive plants in the world, negatively impacting agricultural and natural resources in 73 different countries throughout Africa, Asia, Europe, New Zealand, Oceania and the Americas1-2. Cogongrass forms rapidly-spreading, monodominant stands that displace a large variety of native plant species and in turn threaten the native animals that depend on the displaced native plant species for forage and shelter. To add to the problem, an ornamental variety [I. cylindrica var. koenigii (Retzius)] is widely marketed under the names of Imperata cylindrica 'Rubra', Red Baron, and Japanese blood grass (JBG). This variety is putatively sterile and noninvasive and is considered a desirable ornamental for its red-colored leaves. However, under the correct conditions, JBG can produce viable seed (Carol Holko, 2009 personal communication) and can revert to a green invasive form that is often indistinguishable from cogongrass as it takes on the distinguishing characteristics of the wild-type invasive variety4 (Figure 1). This makes identification using morphology a difficult task even for well-trained plant taxonomists. Reversion of JBG to an aggressive green phenotype is also not a rare occurrence. Using sequence comparisons of coding and variable regions in both nuclear and chloroplast DNA, we have confirmed that JBG has reverted to the green invasive within the states of Maryland, South Carolina, and Missouri. JBG has been sold and planted in just about every state in the continental U.S. where there is not an active cogongrass infestation. The extent of the revert problem in not well understood because reverted plants are undocumented and often destroyed.
Application of this molecular protocol provides a method to identify JBG reverts and can help keep these varieties from co-occurring and possibly hybridizing. Cogongrass is an obligate outcrosser and, when crossed with a different genotype, can produce viable wind-dispersed seeds that spread cogongrass over wide distances5-7. JBG has a slightly different genotype than cogongrass and may be able to form viable hybrids with cogongrass. To add to the problem, JBG is more cold and shade tolerant than cogongrass8-10, and gene flow between these two varieties is likely to generate hybrids that are more aggressive, shade tolerant, and cold hardy than wild-type cogongrass. While wild-type cogongrass currently infests over 490 million hectares worldwide, in the Southeast U.S. it infests over 500,000 hectares and is capable of occupying most of the U.S. as it rapidly spreads northward due to its broad niche and geographic potential3,7,11. The potential of a genetic crossing is a serious concern for the USDA-APHIS Federal Noxious Week Program. Currently, the USDA-APHIS prohibits JBG in states where there are major cogongrass infestations (e.g., Florida, Alabama, Mississippi). However, preventing the two varieties from combining can prove more difficult as cogongrass and JBG expand their distributions. Furthermore, the distribution of the JBG revert is currently unknown and without the ability to identify these varieties through morphology, some cogongrass infestations may be the result of JBG reverts. Unfortunately, current molecular methods of identification typically rely on AFLP (Amplified Fragment Length Polymorphisms) and DNA sequencing, both of which are time consuming and costly. Here, we present the first cost-effective and reliable PCR-based molecular genotyping method to accurately distinguish between cogongrass and JBG revert.

A PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica

We provide a cost-effective and rapid molecular genotyping protocol that employs variety-specific PCR primers that target DNA sequence differences within the chloroplast trnL-F spacer region to differentiate between varieties of Imperata cylindrica (cogongrass) that cannot be distinguished by morphology alone. These varieties include the federally listed noxious weed, cogongrass and closely-related, wide-spread ornamental variety, I. cylindrica var. koenigii (Japanese blood grass).

Wild-tip ve Süs Çeşitler arasında ayırt A PCR tabanlı Genotiplendirme Yöntemi Imperata cylindrica</em

Wild-type I. cylindrica (cogongrass) is one of the top ten worst invasive plants in the world, negatively impacting agricultural and natural resources in ...

Repeatable Stair-step Assay to Access the Allelopathic Potential of Weedy Rice (Oryza sativa ssp.)

Weed competition contributes significantly to yield losses in cropping systems worldwide. The evolution of resistance in many weed species to continuously applied herbicides has presented the need for additional management methods. Allelopathy is a physiological process that some plant species possess that provide the plant with an advantage over its neighbors. Allelopathic crop varieties would be equipped with the ability to suppress the growth of surrounding competitors, thus reducing potential yield loss due to weed interference. This paper focuses on the construction and operation of a stair-step assay used for the screening of the allelopathic potential of a donor species (Oryza sativa) against a receiver weed species (Echinochloa crus-galli) in a greenhouse setting. The structure described in this paper serves as a stand for the plant samples and incorporates a timed watering system for the accumulation and distribution of allelochemicals. Allelochemicals produced by the plant roots are allowed to flow downward through a series of four pots separately into a collection tank and recycled back to the top plant through electric pumps. This method of screening provides an avenue for the allelochemicals from the donor plant to reach receiver plants without any resource competition, thus allowing quantitative measurement of the allelopathic potential of the selected donor plant. The allelopathic potential is measurable through the height reduction of the receiver plants. Preliminary screening data for the effectiveness of this method demonstrated height reduction in the receiver species, barnyardgrass (E. crus-galli), and thus the presence of allelopathic residues from the donor plant, weedy rice (Oryza sativa).

Repeatable Stair-step Assay to Access the Allelopathic Potential of Weedy Rice Oryza sativa ssp.

Allelopathy has shown promise as a useful supplemental weed control strategy in cropping systems. To determine the allelopathic potential of a desired plant specimen, a stair-step screening method is provided.

Weedy Rice Allelopathic Potansiyeli Erişim Tekrarlanabilir Merdiven adım Assay(Oryza sativa ssp.)

Weed competition contributes significantly to yield losses in cropping systems worldwide. The evolution of resistance in many weed species to continuously ...

Farklı Ot Türlerinin Sağlam Herbisit Direnci Testi için Protokoller

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