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High‐throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli
Plant Pathology ( IF 2.7 ) Pub Date : 2020-04-03 , DOI: 10.1111/ppa.13177
Yu Lu 1 , Loïc Deblais 2, 3 , Gireesh Rajashekara 2 , Sally A. Miller 3 , Yorsa A. Helmy 2 , Haijun Zhang 1 , Ping Wu 1 , Yanhong Qiu 1 , Xiulan Xu 1
Affiliation  

Acidovorax citrulli is a seed‐transmitted gram‐negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small‐molecule libraries containing 4,952 compounds were selected for high‐throughput screening against A. citrulli wild‐type strain Xu3‐14 by evaluation of growth inhibition. One hundred and twenty‐seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed‐to‐seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed‐to‐seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli‐specific bactericide.

中文翻译:

高通量筛选揭示小分子调节剂抑制酸食瓜

Acidovorax citrulli 是一种通过种子传播的革兰氏阴性细菌,可导致全球西瓜和甜瓜生产的经济产量大幅下降。通过生长抑制评估,选择了包含 4,952 种化合物的四个小分子文库,用于针对 A. citrulli 野生型菌株 Xu3-14 的高通量筛选。127 个分子(2.5% 的命中率)在 100 μM 下被鉴定为对 A. citrulli 具有杀菌或抑菌作用。二次筛选表明,27 种候选化合物对 A. citrulli Group II 菌株比 Group I 菌株(使用重复元件 PCR 分类)更有效。几种化合物对其他病原菌有抑制作用,包括 Clavibacter michiganensis subsp。michiganensis 和 Xanthomonas campestris pv。油菜,但不影响植物有益菌荧光假单胞菌和枯草芽孢杆菌的生长。超过一半的化合物不抑制拟南芥或西瓜种子的萌发。通过将每种化合物应用于接种的西瓜种子并评估幼苗感染来评估小分子对 A. citrulli 种子到幼苗传播的影响。九种化合物被选择用于进一步研究,因为它们降低了幼苗感染的百分比,并在二次筛选中获得了对它们的特异性、敏感性和植物毒性的综合评分。选择了五种最好的化合物(噻吩菌素、那氟沙星、哌啶酸、环吡酮和吡硫锌)进行温室试验,发现它们可以有效地减少 A. 人工和自然感染的种子中的瓜。这些排名前五的化合物为未来开发柑橘类杆菌特异性杀菌剂奠定了基础。
更新日期:2020-04-03
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