协议的鲁棒抗除草剂试验在不同杂草
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
除草剂是最广泛使用的杂草控制措施，占全球植物保护市场1高达50％。他们是相对便宜的工具，避免劳动密集和费时的土壤耕作活动，并最终导致成本效益，安全和有利可图的粮食生产2。然而，巨大的物候和遗传变异存在于许多杂草，对除草剂的使用过度依赖在一起，经常导致抗除草剂杂草种群的选择。选择性除草剂的一个非常具体的目标代谢引进3-5急剧增加的阻力案件的数量多年来。到目前为止，世界各地的240种杂草（双子叶植物140和100单子叶植物）产生了抗性行动（SOA）4不同的除草剂站点。这是一个大问题杂草管理，更一般的可持续作物生产。
e_content“>早检测电阻的基础上，可靠的试验，在温室中频繁地进行，是管理除草剂抗性杂草的关键步骤。根据该目标，精度，时间和可利用的资源所需的水平不同的方法已被开发，如以及考虑6-12的杂草种类。然而，当确认一个新的杂草生物型的电阻状态是必需的（ 即，一组个人共享几个生理特征，包括生存属于一种或多种除草剂的能力在通常将控制它们的剂量用于特定基），一个强大的整株植物生物测定需要在受控环境4,11进行。一个生物型是刚刚一种除草剂抗性很少。因此，每个生物型的特征在于一定的阻力图案，SOA的即，数量和类型的除草剂是抗性，并且由给定的阻力级别每个除草剂13。早期的和可靠的决心交叉或多重耐药5的格局，14是现场管理性的重要。
值得一提的是，抗除草剂无关与自然容忍对某些除草剂， 例如，双子叶植物与ACC酶抑制除草剂，单子叶植物与2,4-D， 问荆对一些杂草展览草甘膦。
本文提出了一种可靠的方法来测试假定采样除草剂在控制通过除草剂（S）差已经报道领域耐生物型。相关的变体，以相对于所涉及的杂草种类的标准协议被提出。在仅使用一种除草剂剂量15或治疗种子在培养皿8根据任一整株植物生物测定法替代的技术/协议的优点都涉及到更高reliability和推断，因为在实验中列入两个除草剂剂量的阻力水平的可能性。然而，对于常规的电阻测试中，相同的方法可以在仅一种除草剂剂量施加，所以降低了成本。
以及允许确认电阻状态的，可以同时用于优化下面研究步骤和/或制定完善的抗性管理策略获得的信息。
Protocol
Representative Results
Discussion
内协议的几个步骤是抗除草剂的人口的成功关键评估：1）种子应当收集成熟时从幸存的除草剂处理（S）的植物。在母株的种子成熟是至关重要的，以避免在种子发芽后的困难; 2）种子的存放建议避免霉菌，防止发芽的扩散; 3）苗应在合适的生长阶段来处理，所报告的除草剂包装的标签上。必须小心，使得所处理的植物已经大致达到相同的生长阶段; 4）将除草剂溶液应准备并准确处理，以便苗用?…
Disclosures
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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