State Key Laboratory of Agricultural Microbiology, College of Life Sciences, Huazhong Agricultural University, 430070, Wuhan, Hubei Province, China.
Center for Advances in Vaccinology & Biotechnology, University of Baluchistan, Quetta, Pakistan.
Mol Biol Rep. 2022 Jul;49(7):6313-6324. doi: 10.1007/s11033-022-07439-5. Epub 2022 May 9.
Plant-parasitic nematodes (PPNs) are severe threats to agricultural yields and continue to be challenging to treat in several crops worldwide. Microbial-based control has been suggested as a better alternative to chemical control. In this study, we aimed to identify and characterize nematicidal virulence factors of a common phytopathogenic bacterium, Pseudomonas syringae, mainly focusing on the nematicidal and suppressive activities of an NlpC/P60 family peptidase, namely, Peptidase03, against the model nematode Caenorhabditis elegans and an agriculturally important PPN, Meloidogyne incognita.
Genome-wide virulence factor prediction of the P. syringae wild-type strain MB03 revealed numerous nematode pathogenic determinants. We selected 11 predicted nematicidal genes for cloning and induced expression in an Escherichia coli expression system and then performed comparative nematicidal bioassays on the model nematode C. elegans. The recombinant strain expressing Peptidase03 showed the highest level of toxicity against C. elegans, with 75.9% mortality, compared to the other tested strains. Purified Peptidase03 showed significant toxicity against C. elegans and M. incognita, with half lethal concentration (LC) values of 147.9 µg/mL and 211.50 µg/mL, respectively. We also demonstrated that Peptidase03 could damage the intestinal tissues of C. elegans and exhibit detrimental effects on its growth, brood size, and locomotion.
The Peptidase03 protein from P. syringae MB03 had significant nematicidal and suppressive activities against C. elegans and M. incognita, thereby showing potential for the development of an effective PPN-controlling agent for use in agricultural practice.
植物寄生线虫(PPNs)对农业产量构成严重威胁,在全球范围内的几种作物中仍然难以防治。微生物防治被认为是化学防治的更好替代品。在这项研究中,我们旨在鉴定和表征一种常见植物病原菌丁香假单胞菌的杀线虫毒力因子,主要关注 NlpC/P60 家族肽酶(即肽酶 03)对模式线虫秀丽隐杆线虫和农业上重要的 PPN 南方根结线虫的杀线虫和抑制活性。
丁香假单胞菌野生型菌株 MB03 的全基因组毒力因子预测揭示了许多线虫致病决定因素。我们选择了 11 个预测的杀线虫基因进行克隆,并在大肠杆菌表达系统中诱导表达,然后在模式线虫秀丽隐杆线虫上进行比较杀线虫生物测定。与其他测试菌株相比,表达肽酶 03 的重组菌株对秀丽隐杆线虫表现出最高的毒性,死亡率为 75.9%。纯化的肽酶 03 对秀丽隐杆线虫和南方根结线虫表现出显著的毒性,半数致死浓度 (LC) 值分别为 147.9 µg/mL 和 211.50 µg/mL。我们还证明肽酶 03 可以损伤秀丽隐杆线虫的肠道组织,并对其生长、产卵量和运动能力产生不利影响。
丁香假单胞菌 MB03 的肽酶 03 蛋白对秀丽隐杆线虫和南方根结线虫具有显著的杀线虫和抑制活性,因此具有开发用于农业实践的有效 PPN 控制剂的潜力。