UK Centre for Ecology & Hydrology, Benson Lane, Crowmarsh Gifford, Wallingford, UK.
Warwick Crop Centre, School of Life Sciences, Wellesbourne Campus, The University of Warwick, Warwick, UK.
Pest Manag Sci. 2020 Jul;76(7):2513-2524. doi: 10.1002/ps.5794. Epub 2020 Mar 12.
Bioassays evaluating entomopathogenic fungi (EPF) isolates for effective microbial control of whitefly are a fundamental part of the screening process for bioprotectants, but development of repeatable, robust bioassays is not straightforward. Currently, there is no readily available standardised method to test the efficacy of EPF on whitefly. Here, we describe the calibration and use of a spray tower to deliver a standardised protocol to assess EPF activity; the method was validated using 18 EPF from four genera in tests against greenhouse whitefly, Trialeurodes vaporariorum (Westwood).
At 138 kPa, the sprayer delivered 0.062 mL mm (620 L ha ) and an even deposition of spray across the central 1590 mm of the spray area. Average conidial deposition for all EPF was 252 conidia mm and equivalent to 2.5 × 10 conidia ha at an application concentration of 1 × 10 conidia mL . Conidial deposition of a test Beauveria bassiana suspension increased with increasing application concentration. Egg laying by T. vaporariorum adults was restricted to 177 mm using clip cages specifically designed to ensure that third-instar T. vaporariorum received a uniform spray coverage. Nymphs occupied 373 ± 5 mm of the leaf after migrating during the first instar. Average T. vaporariorum mortality totaled 8-89% 14 days after application of 1 × 10 conidia mL of each EPF isolate.
Combining the calibrated sprayer and bioassay method provides a reliable, standardised approach to test the virulence of EPF against whitefly nymphs. This laboratory-based assay is affordable, replicable and allows the user to alter the dose of conidia applied to the target.
评估杀虫真菌(EPF)分离物对白粉虱有效微生物控制的生物测定是生物保护剂筛选过程的基本组成部分,但开发可重复、稳健的生物测定并不简单。目前,尚无现成的标准化方法来测试 EPF 对白粉虱的功效。在这里,我们描述了喷雾塔的校准和使用,以提供一种标准化方案来评估 EPF 活性;该方法使用来自四个属的 18 种 EPF 在温室白粉虱(Trialeurodes vaporariorum(Westwood))测试中得到了验证。
在 138 kPa 下,喷雾器输送 0.062 毫升毫米(620 升公顷),在喷雾区域的中心 1590 毫米范围内均匀沉积喷雾。所有 EPF 的平均分生孢子沉积量为 252 个分生孢子毫米,相当于应用浓度为 1×10 个分生孢子毫升时的 2.5×10 个分生孢子公顷。测试的球孢白僵菌悬浮液的分生孢子沉积量随着应用浓度的增加而增加。专门设计的夹笼将成虫产卵限制在 177 毫米以内,以确保三龄白粉虱幼虫获得均匀的喷雾覆盖。在第一龄期间迁移后,若虫占据 373±5 毫米的叶片。在应用每毫升 1×10 个分生孢子的各 EPF 分离物 14 天后,白粉虱幼虫的平均死亡率总计为 8-89%。
结合校准的喷雾器和生物测定方法为测试 EPF 对白粉虱若虫的毒力提供了一种可靠、标准化的方法。这种基于实验室的测定方法价格实惠、可重复,并且允许用户改变应用于目标的分生孢子剂量。