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. 종자 시료 채취 및 보관 <ol><li> 불리한 기후 조건 또는 품질이 낮은 제초제 처리로 인해하지 즉 부당한 가난한 제초제 성능을 재배 필드, 모니터링합니다. </li><li> 한 번에 한 종의 종자 샘플을 수집하고 고유 한 코드를 할당합니다. 성숙한 종자는 일반적으로 제초제 처리 (들)을 살아남은 식물에서 작물 수확하기 전에 수집됩니다. 씨앗이 때 어머니 식물에 의해 성숙 창고 경우 적시에 모니터를 관찰합니다. </li><li> 종, 수집 날짜, GPS 좌표, 지방 자치 단체, 농부의 이름, 필드 크기, 감염 수준, 작물, 제초제 (들) 시즌 동안 사용 및 필드의 역사적 기록의 할당 된 고유 코드, 이름을 나타내는 각 샘플에 대한 양식을 작성 . </li><li> 필드 침략의 대표 적어도 30 무작위로 선택된 식물에서 종자를 수집합니다. 종자 샘플이 적어도 5,000 성숙한 씨앗이 포함되어 있는지 확인합니다. 의무적 교잡 잡초 종(예를 들어, 종 Lolium. 또는 Amaranthus 종은.), 5000 (11) 주위에 종자의 수를 유지하면서 10-15 식물의 수를 감소시킨다. </li><li> 잡초의 패치 (헥타르 이상) 넓은 지역에 흩어져있는 것처럼 다른 제초제 저항성 생물 형을 선택하는 필드를 하위 샘플. </li><li> 할당 된 고유 코드로 표시된 봉인되지 않은 종이 봉투에 보관 씨앗. </li><li> 수분이 증발하지만 차 휴면 유도를 방지하기 위해 높은 온도 (즉, 태양 아래 차를 떠나 피) 또는 극단적 인 온도 변화에 씨앗을 노출되지 않도록 할 수 있습니다. </li><li> (왕겨, 드 선체 씨앗 등을 제거)하고 건조한 장소에 상온에서 보관 청소. 첫 번째 저항 시험을 수행 한 후, 진공 밀봉 된 비닐 봉지에 바람직하게는 4 ° C에서 어두운 방에 오랜 시간 동안 씨앗을 저장합니다. 이러한 방식 씨앗 상당히 긴 시간 동안 그들의 생존을 유지. </li></ol> 참고 : 종자 휴면 적응과 농업 생태계에 지속 잡초를 할 수있는 유연하고 효율적인 메커니즘을 제공합니다. 종자 발아를 휴면을 중단하고 허용하기 위해 다른 프로토콜 즉 잡초 종, 수면 (16)의 종류에 따라 사용할 수있다. 휴면을 제거하는 세 가지 방법이 있습니다 : <ol><li> 춘화 참고 :. 예를 들어, Amaranthus의 retroflexus, 명아주 앨범, Lolium 종, 귀리 fatua, 폴리 고눔 여뀌 속 : 동시 발아 및 유묘 출현, 일주일에 몇 일에 이르기까지 종자 춘화의 기간을 얻으려면 많은 종에서 생리적 휴면을 제거하기 위해 필요 , Phalaris는 17 ~ 19 paradoxa. 최대 15 일 이상 기간이 Schoenoplectus mucronatus 위해 30 일 개양귀비, 방동사니의 difformis 및 Ammania의 coccinea 필요하고 <suP&gt; 20. <ol><li> 플라스틱 접시에 약간의 탈 물을 넣습니다. 초과를 제거, 필터 종이의 두 층을 잘라 물을 흡수. 종이에 공기 건조 씨앗을 놓습니다. 시간이 필요한 기간 동안 4 ° C에서 냉장고에 플라스틱 접시를 전송합니다. </li></ol></li><li> 혹평 주 : 일부 잡초 종 인해 기계적 휴면 즉 종피 특성에 다른 것보다 더 발아 분해성이며, 21 발아 황산을 사용하여 화학적 혹평의 사용을 필요로한다. <ol><li> 진한 황산 (95~98%)와 비커를 준비합니다. 물이 가득 비커를 준비합니다. 부직포 봉투에 씨앗을 넣습니다. </li><li> 진한 황산에 각각 20 분 또는 5 분 동안 예를 들어, Echinochloa 종. 또는 수수 halepense 씨앗을 만끽 해보세요. </li><li> 핀셋을 사용하여 비커에서 봉투를 타고 물이 가득 비커에 넣어. 열기봉투, 작은 소쿠리에 씨앗을 넣고 철저하게 흐르는 물을 씻어. </li></ol> 변종 : A. 질경이-aquatica는 다른 프로토콜을 필요로 22, 23 <ol><li> 클로로포름의 2 분의 씨앗을 만끽. 탈 이온수와 씨앗을 씻어 흡수 종이를 건조. 5 분 동안 80 % 황산 속에서 종자 덩크. </li><li> 작은 소쿠리에 씨앗을 넣고 철저하게 흐르는 물을 씻어. </li></ol></li><li> 수확 후 종자의 성숙 참고 : 다른 잡초 종의 종자에 관계없이 휴면을 중단하는 데 사용되는 방법, 만기 후 몇 개월 동안 전혀 발아하지 않습니다. <ol><li> RT와 낮은 습도에서 적어도 3~4개월의 기간 동안 씨앗을 저장 한 후 휴면 파괴 (예를 들어, 오리 자 사티 VAR. sylvatica 또는 피 속 rhoeas)에 대해 위의 프로토콜을 따르십시오. </li></ol></li></ol> 3. 종자 발아 <ol><li> 장소 씨는 PLAS에 발아한다0.1 %의 질산 칼륨 (KNO 3)와 0.6 % (M / V) 한천을 포함 TIC 요리 추가 : <ol><li> 탈 이온수를 사용하여 0.6 % + 0.1 %의 KNO 3에 한천 용액을 준비한다. 전자 레인지에서 용해한. </li><li> 플라스틱 접시에 한천 용액을 붓고. 기판을 냉각 한 후 씨앗에 넣어. </li></ol> 변형 토양은 플라스틱 접시에 기질로서 사용될 수있다. 이 연습은 Echinochloa 종에 특히 효율적입니다., S. halepense 및 Lolium 종. </li><li> 각각의 잡초 종을위한 최적의 조건에 따라 빛과 온도 조건을 일주일 정도 발아 캐비닛에 플라스틱 접시를 놓습니다. 가장 겨울 종의 경우, 온도 범위는 25분의 15 ° C의 밤 / 낮의 15-30 μmol m -2 초 -1의 광합성 광자 플럭스 밀도 (PPFD)을 제공하는 네온 튜브 12 시간 광주 기이다. 여름 많은 종의 경우, 온도 범위는 15/30 ° C의 밤 / 일이다. 변형 : S이러한 S. 등 오메 종, halepense, 열처리가 필요합니다. S의 따라서 혹평 후, 씨앗 정상적인 조건에서 다음 삼일 발아 캐비닛에 사흘 동안 24 ° C에서 45 ° C와 20 시간에서 4 시간의주기를, 그리고 : halepense는 다음과 같은 조건을 실시한다. </li></ol> 4. 모종 이식과 성장 <ol><li> 이식 표준 포팅 믹스로 가득 플라스틱 트레이 (325 X 265 X 95mm)에 15-20 모종 (60 % 미사 질 양토, 15 % 모래, 15 % agriperlite 10 % 이탄 - 부피). 주 : 이식 대신 직접 파종, 제초제 처리의 성능을 최적화하는 중요한 전제 조건 인, 동일한 성장 단계에서 식물의 균일 스탠드 얻을 수있다. </li><li> 수와 진보적 인 트레이 번호를 복제, 인구 코드, 제초제가 테스트중인 &lt;: 고유 식별에 대한 모든 정보를 포함하는 바코드 각 트레이를 식별합니다./ 리&gt; <li> 또는 필드의 용량이 거의 기판을 유지하기 위해 가열 된 물을 식물, 온실에서 찾는 트레이. 주 : 성장 온도가 잡초 종류에 따라 달라집니다. 종종 시험은 가을 / 겨울 / 봄에 완료, 그래서 빛이 약 150 μmol 분 -2 초 PPFD을 제공하는 400 W 메탈 할라이드 램프, -1과 12 시간의 광주 (24), (19). 여름 잡초 종을 사용하여 보충 C 4 광합성주기는 일반적으로 더 높은 빛의 강도를 필요로하기 때문에 시험은 늦은 봄 - 여름에서 수행하거나 보충 빛의 세기가 약 400 μmol 분 -2 초 -1 14 시간의 광주에있다된다. </li><li> 패 디 쌀, 예를 들어, A를 감염시키고 일부 잡초 종에 대해 서로 다른 프로토콜을 사용하여 질경이-aquatica, S. mucronatus와 C 22에 기술 된 바와 같이 difformis. <ol><li> 24 라운드 세포 (55mm 직경, 64mm 깊이) FIL와 폴리스티렌 트레이에 모종을 이식60 % 미사 질 양토, 30 %의 모래와 (부피 기준) 10 % 이탄으로 이끌었다. </li><li> 12cm의 트레이 깊은 플라스틱 용기는 물이 가득하고 (그림 1) 부동 방지하기 위해 나사 스테인리스 봉에 의해 아래로 battened을 설정합니다. </li><li> 토양 표면의 레벨 아래 1~2cm에서 용기의 수위를 유지하고 조류의 증식을 방지하기 (10 ~ 12 L의 물을 포함)은 각 컨테이너에 황산구리 1.5 g을 추가한다. </li></ol></li></ol> 5. 제초제 트리트먼트 <ol><li> 사전 출현 제초제와 치료 : <ol><li> 발아 캐비닛에 3 일 정도는 3 장에서 설명한대로 한 후, 위에 설명 된 기판을 포함하는 플라스틱 트레이에 발아 씨앗을 이식 및 토양의 층 (약 1cm) 커버. 이 모종 제초제 효과보다는 과도한 매몰 깊이로 인해 나타날되지 않도록하기 위해서 중요한 공정이다. </li><li> 필드 CAPA에 기판을 가지고물이 가득 접시에, 바닥에 약간의 구멍이 트레이를 배치하여 도시. </li><li> 어느 날 이식 후 사전 출현 제초제 (25) 트레이를 취급합니다. </li><li> 접시에서 모세관 현상에 의해 위와​​ 아래 모두에 필요한 물을 첨가하여 상기 기판 또는 근접장 용량을 유지. 이 절차는 좋은 치료 효과에 대한 (즉, 발아 씨앗이있는 곳) 적절한 깊이에 제초제의 영속성을 선호한다. </li></ol></li><li> 포스트 출현 제초제와 치료 : <ol><li> 그들은 2-3 잎 단계에 도달 할 때 식물 스프레이 (즉, 성장 단계 확장 BBCH 성장 규모 (26)의 12 ~ 13). </li><li> 치료 다음 날부터 시작, 잡초 종의 물 요구 사항 및 Echinochloa 종의 계절 (예에 따라 관개 시스템을 설정합니다.이 9 오전 9 시부 터 일정한 간격으로, 3 분 동안 하루에 4 번 물을 제공합니다 PM). 물은 DIS입니다자동 스프링클러 관개 시스템을 사용하여 tributed. 변종 : 제초제는 식물 무대 BBCH 14-21에 적용됩니다. </li></ol></li><li> 제초제 준비 및 배포. 주 : (전후 출현) 모든 제초제 두 용량으로 권장 활성제로 상업적 제제로서 적용되어 추천 필드 도즈 (1X) 및 세 배 (3X)있다. <ol><li> 필요한 경우, 라벨 지시에 따라 대량의 계면 활성제 용액을 제조 하였다; 최종 농도는 일반적으로 최종 부피의 백분율 (예를 들어, 0.3 %)로 표현하거나 볼륨으로 단위 면적 (예를 들면, 1 L의 HA-1)별로 분배 될. </li><li> 활성 성분의 적절한 농도를 유지하기 위하여 제초제 (용질) 용액 용 용매로서 계면 활성제 용액을 사용한다. 첫 번째 제초제 솔루션을 준비합니다 (3 배). 상업적 제품의 수량이 계면 활성제 용액에 용해 (또는 경우 탈 이온수으로 계산할계면 활성제는 다음의 방정식을 사용하여)가 필요하지 않다 : 용량 허브 = [(용량 필드 X 선량 최대) × V 핀] / V 델 여기서 용량 허브 = 제초제 용량 (㎖), 용량 필드 = 제초제 필드 용량 (하 ㎖를-1), 용량 최대 = 최대 용량 제공, V 지느러미는 = 솔루션 (L)의 최종 부피는 V 델은에 의해 전달 볼륨을 = 벤치 분무기 (L 하 -1). </li><li> 덜 집중 하나 (1 배)을 제조 : (1, V / V 2) 제초제 솔루션 배 희석. 이 절차는 계량 또는 제초제를 피펫 팅 할 때 실수의 가능성을 감소시킨다. 볼륨 단위 영역 (L 헥타르 -1) 당 분산로서 제초제 용액 농도가 발현된다. </li><li> 낮은 제초제 량 (1X) 치료의 순서를 시작합니다. 이러한 방식으로 동일한 두 제초제먼트 사이 분무 캐비넷 세척 할 필요가 없다. </li><li> 제초제 solutio 배포N 세 평면 팬 (확장 범위) 유압 노즐 장착 붐 215 kPa의 압력에서 300 L의 HA-1 (± 1 %), 전달 정밀도 벤치 분무기, 0.75 m 초 -1의 속도를 사용하여 . </li><li> 제초제는 표백제를 1 %를 사용하여 변경 될 때 두 번 분사 캐비닛을 씻으 (v / v)의 다음 씻어. 변종 : 글 리포 세이트 200 L 하 -1 (27)의 스프레이 볼륨에 적용됩니다. 참고 : 특별한 관심이 높은 생물 제초제가, 같은 설 포닐 우레아의 sulfometuron 또는 flazasulfuron가 사용되는 경우 지불해야합니다. 후자의 경우에는 암모니아로 한 표백제 용액으로 세척하고 다른 작업을 수행 (2.5 % v / v)의 물로 조심 린스. </li></ol></li></ol> 6. 수집 및 데이터 분석 <ol><li> 각 트레이를 자동 식별 바코드 판독기를 통해, 치료뿐만 아니라 비주얼 기준 매스 (VEB)을 생존 식물의 수를 기록한다. 식물은 엉덩이입니다그들이 상관없이 색상이나 다른 모양의 활성 증가를 보여 경우 죽은 것으로 ESSID와. <ol><li> 테스트 제초제에 따라 세 사주 치료 (와트) 후 평가를 (예를 들어, ACCase 억제제 세 와트 및 루게릭 병 억제제 또는 글 리포 세이트에 대한 네 와트). </li><li> 즉, 취약 인구 (S 확인) 모든 실험에서, 결코 또는 드물게 제초제로 처리하지 않는 사이트에서 수집 된 인구를 포함하여 일반적으로 치료 효과를 평가합니다. </li><li> 처리 된 식물들의 수의 비율로 익스프레스 식물 생존율, (두 반복 실험의 평균값) 다만 제초제 처리 전의 계산하고, 평균값에 따라 표준 에러 (SE)를 계산한다. </li><li> 처리와 같은 인구 (25), (28)의 확인을 처리하지 사이에 VEB 식물 바이오 매스를 시각적으로 비교를 통해 얻을 수있다. 점수를, 0 (하지 처리 검사에 비해) 제초제의 영향을받지 않는 식물 (10)에 이르기까지 언제식물이 명확하게 죽은, 각 처리 트레이로 제공됩니다. </li></ol></li><li> S 생존자 이상 제초제 도즈 1X, R에서 20 %로 5 % 내지 원거리 때 식물의 5 % 미만이 제초제 도즈 1X, SR 살아남은 두 제초제 투여 량으로 치료에서 얻어진 결과에 기초하여 네 가지 범주로 돌리다 개체군 생존자 제초제 도즈 1X에서 20 % 이상 및 제초제 도즈 3X 17에서 10 % 이상이있는 경우 식물의 20 % 제초제 도즈 1X 및 RR 살아 남았다. </li></ol>

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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) 종자가 성숙 제초제 처리 (들)을 살아남은 식물에서 수집되어야한다 : 프로토콜 내에서 몇 가지 단계가 인구에 제초제 저항의 성공적인 평가를 위해 중요하다. 어머니 식물 종자의 성숙 나중에 종자 발아에 어려움을 피하기 위해 매우 중요하다; 2) 씨의 적절한 보관은 발아를 방지 할 수 금형의 확산을 방지하는 것이 좋습니다; 제초제 패키지의 라벨에보고 3) 묘목은 오른쪽 성장 단계에서 처리?...

<html> 제초제는 글로벌 식물 보호 시장 (1)의 50 %를 차지, 가장 광범위하게 사용 잡초 측정 한 것입니다. 그들은 노동 집약적이고 시간이 많이 소요되는 토양 재배 관행을 방지하고, 궁극적으로 비용 효율적이고 안전하고 유익한 식품 생산이 결과, 상대적으로 저렴한 도구입니다. 그러나 함께 제초제 사용에 이상 의존 많은 잡초 종의 큰 생물 계절 및 유전 적 다양성의 존재는, 자주 제초제 저항성 잡초 집단의 선택을 초래한다. 매우 특정한 대사 표적 선택적 제초제 도입 3-5 년간 저항 경우의 수를 극적으로 증가하고있다. 현재까지 전 세계적으로 240 잡초 종 (140 dicots 100 외떡잎은) 액션 (SOA) 4의 다른 제초제 사이트에 대한 저항을 진화했다. 이것은 지속 가능한 작물 생산을위한 주요 잡초 관리를위한 관심과 일반적으로 더 많은 것이다. 자주 온실에서 수행 신뢰성 시험에 기초 e_content &quot;&gt; 저항의 조기 발견, 제초제 내성 잡초를 관리하는 중요한 단계이다. 다른 접근법으로, 목표 정확도, 시간 및 이용 가능한 자원의 요구 수준에 따라 개발되어왔다 잘 6-12 간주 잡초 종으로. 그러나, 새로운 잡초 생물 형의 저항 상태의 확인이 (필요한 경우 즉,에 속하는 하나 이상의 제초제 생존 할 수있는 능력 등 여러 가지 생리 학적 특성을 공유하는 사람들의 그룹, 일반적으로 이들을 제어 할 도즈 사용 특정 그룹), 견고한 전체 식물 생물 검정법은 제어 된 환경 4, 11에서 수행 될 필요가있다. 생물 형이 하나의 제초제에 내성이 드물게있다. 각 생물 형 따라서이에 강한 제초제의 특정 저항 패턴, SOA의 즉, 수와 유형을 특징으로, 주어진 저항입니다각 제초제 13 수준. 간 또는 복수의 저항 (5)의 패턴의 조기 판정 및 신뢰성은 14 필드 저항성 관리에 중요하다. 그것은 그 제초제 저항이 자연의 허용 오차와 아무 상관이 언급 할 가치가 일부 제초제, 예를 들어, ACCase 억제 제초제, 단자엽 종 대 2,4-D 대 쌍자엽 종, 속새 arvense 대 대한 몇 가지 잡초 종 전시 글 리포 세이트. 이 논문은 제초제 (들)에 의해 가난한 제어가보고되었다 분야에서 샘플링 추정 제초제 저항성 생물 형을 테스트하기위한 강력한 방법을 제공합니다. 관련된 잡초 종과 관련하여 표준 프로토콜에 관련 변형되게됩니다. 페트리 접시 (8)에 단 하나의 제초제 용량 (15), 또는 치료 씨앗을 사용하거나 전체 공장 생물 검정을 기반으로 다른 기술 / 프로토콜을 통해 장점은 높은 reliab 관련된ility 때문에 실험에서 두 제초제 선량 개재물 저항 레벨을 추정하는 가능성. 그러나, 루틴 저항 테스트를 위해, 동일한 방법이 때문에 비용을 절감 하나만 제초제 용량으로 적용 할 수있다. 뿐만 아니라 저항 상태의 확인을 허용하는대로, 얻어진 정보는 모두 다음의 연구 단계를 최적화 및 / 또는 음향 저항 관리 전략을 고안에 사용될 수있다.

<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.

추정 내성 인구의 저항 상태를 평가하기 위해, 제초제의 효과를 확인하기 위해 분석에서 감수성 검사를 포함하는 기본이다. P. 실시 스크리닝 테스트 결과 속 rhoeas 집단은, 밀 필드를 감염시키고 잡초는, 감수성 검사 (09-36)에와 의심에 강한 하나 (10-91)에 사 후 출현 제초제의 효능에 나타난 그림 2에보고됩니다. 인구 09-36 완전히 하나의 식물, 플로라와 tribenuron 메틸 ...

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.

다른 잡초 종의 강력한 제초제 저항 테스트를위한 프로토콜

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

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14.5K 조회수.  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.

비디오: 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.

플랜트 병원체의 유전자 조작<em&gt; Ustilago maydis</em&gt; 곰팡이 생물과 식물 미생물 상호 작용을 연구하는

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 ...

연구

유전학

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.

절제 잎 분석과 쌍자엽 잡초의 제초제 대사의 요금을 측정

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

환경과학

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.

잔디가 시스템에 양도 잔류 농약을 정량화하기위한 프로토콜

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.

르니포름 선충 저항을 위한 면 유전자형 검열

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.

글리포세이트 기반 제품이 마이크로바이옴에 미치는 잠재적 영향의 정량화

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.

모니터링 살충제 저항 Bioassays

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

생물학

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.

구현<em&gt; 체외</em&gt; 약물 저항 분석법 : 임상 적 저항 메커니즘을 폭로 가능성을 극대화

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

의학

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.

특성화 Herbivore 저항 메커니즘 : Spittlebugs에 Brachiaria 종. 예로

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).

의 야생 - 타입과 장식 품종을 구별하기 PCR 기반 Genotyping 방법 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.

잡초 쌀의 알수 로병성 잠재력에 액세스하는 반복 계단 단계 분석(오리자 sativa ssp.)

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

다른 잡초 종의 강력한 제초제 저항 테스트를위한 프로토콜

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