別の雑草種の堅牢な除草剤抵抗性試験のためのプロトコル
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
除草剤はグローバル植物保護市場1の最大50％を占め、雑草防除対策を最も広く使用されています。彼らは労働集約的で時間のかかる土壌栽培慣行を回避するには、比較的安価なツールであり、最終的に費用対効果の高い、安全かつ収益性の高い食糧生産2をもたらします。しかし、一緒に除草剤の使用上の過度の依存を持つ多くの雑草種に大きな生物季節と遺伝的変異の存在は、しばしば除草剤抵抗性雑草集団の選択をもたらします。非常に特定の代謝ターゲット3-5と選択性除草剤の導入が飛躍的に年間で抵抗症例数が増加しています。現在までに、世界中の240雑草種（140双子葉植物と単子葉植物は100）アクション（SOA）4の異なる除草剤サイトに対する抵抗性を進化させてきました。これは、持続可能な作物生産のための雑草管理と一般的に多くのための主要な関心事です。
頻繁に温室で行う信頼性試験、に基づいてe_content ">抵抗の早期検出は、除草剤抵抗性雑草を管理するための重要なステップである。別のアプローチがよう、目的、精度、時間と利用可能なリソースの要求レベルに応じて開発されていますよく6-12を考え雑草種とし て。しかし、新しい雑草バイオタイプの抵抗状態の確認が（必要とされる場合、すなわち、に属する1つまたは複数の除草剤に耐える能力を含むいくつかの生理学的特性を共有する個人のグループ、通常、それらを制御することになる用量で使用される特定の基）、強固な全植物バイオアッセイは、制御された環境4、11で実行される必要があります。バイオタイプはただ1つの除草剤に耐性めったにあります。各生物型は、従って、一定の抵抗パターンを特徴とする、除草剤、SOAの、すなわち、数と種類は、それは、与えられた抵抗によってに耐性であります各除草剤13のレベル。クロスまたは複数の抵抗5のパターンの早期かつ信頼性の高い決意、14は、圃場抵抗性管理のために重要です。
それは、その除草剤抵抗性は自然寛容とは何の関係もない言及する価値があるいくつかの除草剤、 例えば、ACCアーゼ阻害除草剤、単子葉植物種対2,4-D対双子葉植物種、 スギナarvense対に向けていくつかの雑草種の展示グリホサート。
本稿では、除草剤（複数可）によって制御不良が報告されていた分野でサンプリング推定される除草剤抵抗性生物型をテストするための堅牢なアプローチを提示します。関連する雑草種に関連する標準的なプロトコルに関連する変異体が提示されています。唯一の除草剤の投与量15を使用して全植物バイオアッセイのいずれかに基づいて、代替技術/プロトコル経由での利点、または8が高いreliabに関連しているペトリ皿に種子を処理しますilityとための実験における2つの除草剤の投与量を含めることの抵抗レベルを推定する可能性。しかし、ルーチン抵抗試験のため、同一の方法がそのようにコストを削減し、唯一の除草剤用量で適用することができます。
同様に、抵抗状態の確認を可能にするように、得られた情報は、両方の研究は、以下の工程を最適化し、および/または音響抵抗管理戦略を考案するために使用することができます。
Protocol
Representative Results
Discussion
1）種子が成熟した除草剤処理（複数可）を生き残った植物から収集する必要がありますプロトコル内のいくつかのステップは、集団における除草剤耐性の成功を評価するために重要です。マザー工場の種子の成熟は、後に種子の発芽における困難を回避することが重要です。 2）種子の適切な保管が発芽を妨げるカビの増殖を回避することをお勧めします。除草剤パッケージのラベルに報告…
Divulgazioni
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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