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新型四川链霉菌对古巴尖孢镰刀菌古巴专化型 4 号的生防潜力及其体外和体内的抑菌机制。

Biocontrol potential and antifungal mechanism of a novel Streptomyces sichuanensis against Fusarium oxysporum f. sp. cubense tropical race 4 in vitro and in vivo.

机构信息

Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Hainan Institute for Tropical Agricultural Resources, Haikou, 571101, China.

Ministry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Science, Hainan Normal University, Haikou, 570228, China.

出版信息

Appl Microbiol Biotechnol. 2022 Feb;106(4):1633-1649. doi: 10.1007/s00253-022-11788-3. Epub 2022 Feb 10.

Abstract

Most commercial banana cultivars are highly susceptible to Fusarium wilt caused by soilborne fungus Fusarium oxysporum f. sp. cubense (Foc), especially tropical race 4 (TR4). Biological control using antagonistic microorganism has been considered as an alternative method to fungicide. Our previous study showed that Streptomyces sp. SCA3-4 had a broad-spectrum antifungal activity from the rhizosphere soil of Opuntia stricta in a dry hot valley. Here, the sequenced genome of strain SCA3-4 contained 6614 predicted genes with 72.38% of G + C content. A polymorphic tree was constructed using the multilocus sequence analysis (MLSA) of five house-keeping gene alleles (atpD, gyrB, recA, rpoB, and trpB). Strain SCA3-4 formed a distinct clade with Streptomyces mobaraensis NBRC 13819 with 71% of bootstrap. Average nucleotide identity (ANI) values between genomes of strain SCA3-4 and S. mobaraensis NBRC 13819 was 85.83% below 95-96% of the novel species threshold, and named after Streptomyces sichuanensis sp. nov. The type strain is SCA3-4 (= GDMCC 4.214 = JCM 34964). Genomic analysis revealed that strain SCA3-4 contained 36 known biosynthetic gene clusters of secondary metabolites. Antifungal activity of strain SCA3-4 was closely associated with the production of siderophore and its extracts induced the apoptosis of Foc TR4 cells. A total of 12 potential antifungal metabolites including terpenoids, esters, acid, macrolides etc. were obtained by the gas chromatography-mass spectrometry (GC-MS). Greenhouse experiment indicated that strain SCA3-4 could significantly inhibit infection of Foc TR4 in the roots and corms of banana seedlings and reduce disease index. Therefore, strain SCA3-4 is an important microbial resource for exploring novel natural compounds and developing biopesticides to manage Foc TR4. KEY POINTS: • Strain SCA3-4 was identified as a novel species of Streptomyces. • Siderophore participates in the antifungal regulation. • Secondary metabolites of strain SCA3-4 improves the plant resistance to Foc TR4.

摘要

大多数商业香蕉品种对由土壤传播真菌尖孢镰刀菌古巴专化型(Foc)引起的枯萎病高度敏感,尤其是热带 4 号(TR4)。利用拮抗微生物进行生物防治已被认为是一种替代杀菌剂的方法。我们之前的研究表明,来自干热河谷仙人掌中的土壤的链霉菌 SCA3-4 具有广谱抗真菌活性。在这里,菌株 SCA3-4 的测序基因组包含 6614 个预测基因,G+C 含量为 72.38%。使用 5 个看家基因等位基因(atpD、gyrB、recA、rpoB 和 trpB)的多位点序列分析(MLSA)构建了一个多态性树。菌株 SCA3-4 与 Streptomyces mobaraensis NBRC 13819 形成一个独特的分支,支持率为 71%。菌株 SCA3-4 和 S. mobaraensis NBRC 13819 基因组之间的平均核苷酸同一性(ANI)值低于 95-96%的新种阈值为 85.83%,并被命名为四川链霉菌新种。模式菌株为 SCA3-4(=GDMCC 4.214=JCM 34964)。基因组分析表明,菌株 SCA3-4 含有 36 个已知的次级代谢产物生物合成基因簇。菌株 SCA3-4 的抗真菌活性与铁载体的产生密切相关,其提取物诱导 Foc TR4 细胞凋亡。通过气相色谱-质谱联用(GC-MS)共获得 12 种潜在的抗真菌代谢产物,包括萜类、酯类、酸类、大环内酯类等。温室实验表明,菌株 SCA3-4 能显著抑制香蕉幼苗根部和鳞茎中 Foc TR4 的感染,降低病情指数。因此,菌株 SCA3-4 是探索新型天然化合物和开发防治 Foc TR4 生物农药的重要微生物资源。 关键点: • SCA3-4 菌株被鉴定为链霉菌的一个新种。 • 铁载体参与抗真菌调节。 • SCA3-4 菌株的次生代谢产物提高了植物对 Foc TR4 的抗性。

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