Abstract Aphanomyces euteiches is an important root pathogen of several legume and forage crops worldwide. Disease management is challenging due to resilient oospores that can persist in the soil for long periods of time. Several research groups are working towards incorporating disease resistance into crop cultivars involving large-scale disease resistance screening and, for some crops, the use of molecular markers associated with quantitative trait loci for resistance. The refinement of laboratory protocols for efficient pathogen isolation, reliable and simple zoospore production, and a repeatable indoor screening assay are important tools needed to further research in this area. The standardization of these protocols among research groups would provide an efficient platform for research collaboration. The effect of different media and sporulation induction methods on zoospore production was assessed. The most effective and repeatable method of A. euteiches zoospore production involved culturing and incubation for 4 days on autoclaved wheat leaf segments placed on corn meal-yeast extract-phosphate buffer agar. Colonized wheat leaves were transferred into 100 mL of distilled water in 250 mL flasks and incubated at 100 rpm for 18 h at 24ºC to induce zoospore production. Five zoospore concentrations ranging from 100 to 10 000 zoospores mL−1 were evaluated using susceptible and partially resistant pea and lentil genotypes to optimize zoospore concentration for germplasm screening. The best differentiation of levels of resistance among lentil and pea genotypes was achieved with 1000 zoospores mL−1.

中文翻译：

---

豌豆和扁豆抗性筛选中真丝酵母游动孢子产量和接种浓度的优化

摘要 Aphanomyces euteiches 是世界范围内几种豆类和饲料作物的重要根病菌。由于具有弹性的卵孢子可以在土壤中长期存在，因此疾病管理具有挑战性。几个研究小组正致力于将抗病性纳入作物品种，包括大规模抗病性筛选，对于某些作物，使用与数量性状基因座相关的分子标记进行抗性。改进用于有效分离病原体、可靠和简单的游动孢子生产以及可重复的室内筛选试验的实验室协议是该领域进一步研究所需的重要工具。研究小组之间这些协议的标准化将为研究合作提供一个有效的平台。评估了不同培养基和孢子形成诱导方法对游动孢子产生的影响。最有效和可重复的 A. euteiches 游动孢子生产方法包括在置于玉米粉-酵母提取物-磷酸盐缓冲液琼脂上的高压灭菌小麦叶段上培养和孵育 4 天。将定植的小麦叶子转移到 250 mL 烧瓶中的 100 mL 蒸馏水中，并在 24ºC 下以 100 rpm 的速度培养 18 小时以诱导游动孢子的产生。使用易感和部分抗性豌豆和扁豆基因型评估了 100 到 10 000 个游动孢子 mL-1 的五种游动孢子浓度，以优化用于种质筛选的游动孢子浓度。用 1000 个游动孢子 mL-1 实现了扁豆和豌豆基因型之间抗性水平的最佳区分。