Protokoller for Robust herbicidresistens Afprøvning i forskellige ukrudtsarter
Summary
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.
Abstract
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.
Introduction
Herbicider er de mest flittigt brugt ukrudtsbekæmpelse foranstaltning, der tegner sig for op til 50% for beskyttelse markedet for globale plant 1. De er relativt billige værktøjer, undgå arbejdsintensive og tidskrævende jord dyrkningsmetoder, og i sidste ende resultere i omkostningseffektive, sikker og lønsom fødevareproduktion 2. Men den store fænologiske og genetisk variation til stede i mange ukrudtsarter, sammen med en stor afhængighed af herbicid anvendelse, ofte resulterer i udvælgelsen af herbicidresistente ukrudt befolkninger. Indførelsen af selektive herbicider med en meget specifik metabolisk target 3-5 har dramatisk forøget antallet af resistens sager i årenes løb. Til dato har 240 ukrudtsarter (140 tokimbladede og 100 enkimbladede) i hele verden udviklet resistens over for forskellige herbicider steder i aktion (SOA) 4. Dette er et stort problem for ukrudt og mere generelt for bæredygtig produktion af afgrøder.
e_content "> Tidlig påvisning af resistens, baseret på pålidelige tests, der ofte udføres i et drivhus, er et vigtigt skridt til at håndtere herbicidresistente ukrudt. Forskellige tilgange er blevet udviklet i henhold til de mål, krævede niveau af nøjagtighed, tid og ressourcer til rådighed, som samt ukrudtsarterne betragtes 6-12. Men når der kræves bekræftelse af modstanden status for en ny ukrudt biotype (dvs. en gruppe af individer, der deler flere fysiologiske egenskaber, herunder evnen til at overleve en eller flere herbicider, der tilhører en særlig gruppe anvendes i en dosis, der normalt ville kontrollere dem), skal udføres i et kontrolleret miljø 4, 11 a robust hel plante bioassay.En biotype sjældent resistent over for blot ét herbicid. Hver biotype er derfor karakteriseret ved en vis modstand mønster, dvs, antallet og typen af SOA af herbiciderne er resistent over for, og ved en given modstandniveau til hver herbicid 13. Den tidlige og pålidelig bestemmelse af mønstret af indlægget eller multipel resistens 5, 14 er vigtig for feltet resistenshåndtering.
Det er værd at nævne, at herbicidresistens har intet at gøre med den naturlige tolerance, at nogle ukrudtsarter udstille mod nogle herbicider, fx tokimbladet arter vs. ACCase-hæmmende herbicider, enkimbladede arter vs. 2,4-D, Equisetum arvense vs. glyphosat.
Denne artikel præsenterer en robust tilgang til test formodede herbicidresistente biotyper stikprøven på områder, hvor var blevet indberettet dårlig kontrol af herbicid (r). Relevante varianter til standardprotokoller i forhold til de ukrudtsarter, der er involveret præsenteres. De fordele i forhold alternative teknikker / protokoller baseret på enten hele planten bioassays bruger kun en herbicid dosis 15 eller behandling frø i petriskåle 8 er relateret til højere reliability og muligheden for at udlede modstanden niveau på grund af inddragelsen af to herbicider doser i forsøgene. Men til test rutine modstand, kan anvendes de samme metoder på kun ét herbicid dosis, så at reducere omkostningerne.
Samt ved at lade bekræftelse af resistens status, kan de indsamlede oplysninger bruges til både at optimere de følgende forsknings- trin og / eller udarbejde fornuftige strategier modstand management.
Protocol
Representative Results
Discussion
Flere trin inden for de protokoller er afgørende for en vellykket vurdering af herbicidresistens i en population: 1) frø skal indsamles, når moden fra planter, der havde overlevet ukrudtsmidlet behandling (er). Modning af frø på moderplanten er afgørende at undgå vanskeligheder med frøspiring senere; 2) anbefales det korrekt opbevaring af frø for at undgå spredning af forme, der ville forhindre spiring; 3) kimplanter skal behandles på det rigtige vækststadium, som rapporteret på etiketten af herbicide…
Declarações
The authors have nothing to disclose.
Acknowledgements
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.
Materials
| Paper bags | Celcar SAS | ||
| Plastic dishes | ISI plast S.p.A. | SO600 | Transparent plastic |
| Sulfuric acid 95-98% | Sigma-Aldrich | 320501 | |
| Non-woven fabric | Carretta Tessitura | Art.TNT17 | Weight 17 gr m–² |
| Chloroform >99.5% | Sigma-Aldrich | C2432 | |
| Agar | Sigma-Aldrich | A1296 | |
| Potassium nitrate >99.0% | Sigma-Aldrich | P8394 | |
| Plastic containers | Giganplast | 1875/M | 600 x 400 x 110 mm |
| Plastic trays | Piber plast | G1210A | 325 x 265 x 95 mm |
| Polystyrene trays | Plastisavio | S24 | 537x328x72 mm, 24 round cells (6×4) |
| Copper sulfate | Sigma-Aldrich | 451657 | |
| Agriperlite | Blu Agroingross sas | AGRI100 | |
| Peat | Blu Agroingross sas | TORBA250 | |
| Germination cabinet | KW | W87R | |
| Nozzles | Teejet | XR11002-VK, TP11001-VH | The second type of nozzles are used only for glyphosate |
| Barcode generator | Toshiba TEC | SX4 | |
| Labels with barcode | Felga | TT20200 | Stick-in labels with rounded corners |
| Barcode reader | Cipherlab | 8300-L | Portable data terminal |
| Bench sprayer | – | – | Built in house |
| HERBICIDES INCLUDED IN THE RESULTS: | |||
| Commercial product | Active ingredient | Company | Comments |
| Altorex | imazamox | BASF | |
| Azimut | florasulam | Dow AgroSciences | |
| Biopower | Bayer Crop Science | Surfact to be used with Hussar WG | |
| Dash | BASF | Surfact to be used with Altorex | |
| Granstar | tribenuron-methyl | Dupont | |
| Gulliver | azimsulfuron | Dupont | |
| Hussar WG | iodosulfuron | Bayer Crop Science | |
| Nominee | bispyribac-Na | Bayer Crop Science | |
| Roundup | glyphosate | Monsanto | |
| Trend | Dupont | Surfact to be used with Granstar and Gulliver | |
| Viper | penoxsulam | Dow AgroSciences | |
| Weedone LV4 | 2,4-D | Isagro | |
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