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研究微生物农药申嗪霉素对茎点霉属真菌的抑菌活性及其作用机制

Investigating the antifungal activity and mechanism of a microbial pesticide Shenqinmycin against Phoma sp.

机构信息

State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Huaxi District, Guiyang 550025, China.

State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Huaxi District, Guiyang 550025, China.

出版信息

Pestic Biochem Physiol. 2018 May;147:46-50. doi: 10.1016/j.pestbp.2017.08.014. Epub 2017 Aug 30.

Abstract

Tea white scab (TWS) is a major disease affecting tea trees in mid-elevation regions and often occurs during rainy seasons with low temperatures. This disease is caused by the fungal pathogen Phoma sp. TWS can infect young stems, tender leaves, and tender shoots and lead to the production of low-quality tea. Owing to the absence of an effective control, TWS can result in substantial loss in tea production. In this study, we isolated and identified the pathogen from tea leaves infected by TWS and then evaluated in vitro the antifungal activity of Shenqinmycin, polyoxin, azoxystrobin, oligosaccharins, and tebuconazole against Phoma sp. Our results indicated that Shenqinmycin can inhibit the growth of Phoma sp. mycelia, with the EC value of 0.74μg/mL. After Phoma sp. being incubated in PDB liquid medium with Shenqinmycin, its mycelia were distorted and distended at 1.56μg/mL of minimum inhibitory concentration for 6h. Crucial genes associated with cell redox homeostasis, proteins synthesis, energy metabolism, and cytoskeleton were studied at mRNA and protein levels through RT-qPCR and Nano-LC-MS/MS. The results showed that the genes of 3-phosphate-glyceraldehyde dehydrogenase, citrate synthase, NADH-ubiquinone oxidoreductase subunit (NADH-subunit), ribosomal protein, eukaryotic initiation factor 4A-I, β-tubulin, and α-tubulin were up-regulated. Meanwhile, the genes of formate dehydrogenase (FDH), malate dehydrogenase, mitochondrial heat shock protein, and protein disulfide-isomerase (PDI) were up-regulated at mRNA level but down-regulated at protein level. These results indicated that Shenqinmycin contribute to cell redox homeostasis by up- or down-regulating NADH-subunit, FDH, and PDI.

摘要

茶白星病(TWS)是一种影响中海拔地区茶树的主要病害,常发生在低温多雨季节。这种病害是由真菌病原菌 Phoma sp.引起的。TWS 可感染嫩茎、嫩叶和嫩梢,导致茶叶质量下降。由于缺乏有效的控制措施,TWS 可能导致茶叶产量大幅下降。在本研究中,我们从感染 TWS 的茶树叶片中分离和鉴定了病原菌,然后评估了申嗪霉素、多氧霉素、唑菌胺酯、寡糖素和戊唑醇对 Phoma sp.的体外抑菌活性。结果表明,申嗪霉素能抑制 Phoma sp.菌丝生长,EC 值为 0.74μg/mL。当 Phoma sp.在含申嗪霉素的 PDB 液体培养基中孵育时,其菌丝在 1.56μg/mL 的最低抑菌浓度下 6h 后发生扭曲和肿胀。通过 RT-qPCR 和 Nano-LC-MS/MS 研究了与细胞氧化还原稳态、蛋白质合成、能量代谢和细胞骨架相关的关键基因在 mRNA 和蛋白质水平上的表达。结果表明,3-磷酸甘油醛脱氢酶、柠檬酸合酶、NADH-泛醌氧化还原酶亚基(NADH-亚基)、核糖体蛋白、真核起始因子 4A-I、β-微管蛋白和α-微管蛋白的基因上调。同时,甲酸脱氢酶(FDH)、苹果酸脱氢酶、线粒体热休克蛋白和蛋白二硫键异构酶(PDI)的基因在 mRNA 水平上调,但在蛋白质水平下调。这些结果表明,申嗪霉素通过上调或下调 NADH-亚基、FDH 和 PDI 来维持细胞氧化还原稳态。

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