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从中部墨西哥患有葡萄蔓枯病症状的葡萄藤中分离和酶特性分析真菌菌株。

Isolation and Enzymatic Characterization of Fungal Strains from Grapevines with Grapevine Trunk Diseases Symptoms in Central Mexico.

机构信息

Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las campanas S/N, Col. Las campanas, C. P. 76010, Querétaro, Mexico.

Red Manejo Biotecnológico de Recursos, Instituto de Ecología, A.C., Carretera antigua a Coatepec 351, El Haya, C. P. 91073, Xalapa, Veracruz, Mexico.

出版信息

Curr Microbiol. 2024 May 31;81(7):200. doi: 10.1007/s00284-024-03709-6.

Abstract

Grapevine production is economically indispensable for the global wine industry. Currently, Mexico cultivates grapevines across approximately 28 500 hectares, ranking as the 26th largest producer worldwide. Given its significance, early detection of plant diseases' causal agents is crucial for preventing outbreaks. Consequently, our study aimed to identify fungal strains in grapevines exhibiting trunk disease symptoms and assess their enzymatic capabilities as indicators of their phytopathogenic potential. We collected plant cultivars, including Malbec, Shiraz, and Tempranillo, from Querétaro, Mexico. In the laboratory, we superficially removed the plant bark to prevent external contamination. Subsequently, the sample was superficially disinfected, and sawdust was generated from the symptomatic tissue. Cultivable fungal strains were isolated using aseptic techniques from the recovered sawdust. Colonies were grown on PDA and identified through a combination of microscopy and DNA-sequencing of the ITS and LSU nrDNA regions, coupled with a BLASTn search in the GenBank database. We evaluated the strains' qualitative ability to degrade cellulose, starch, and lignin using specific media and stains. Using culture morphology and DNA-sequencing, 13 species in seven genera were determined: Acremonium, Aspergillus, Cladosporium, Dydimella, Fusarium, Sarocladium, and Quambalaria. Some isolated strains were able to degrade cellulose or lignin, or starch. These results constitute the first report of these species community in the Americas. Using culture-dependent and DNA-sequencing tools allows the detection of fungal strains to continue monitoring for early prevention of the GTD.

摘要

葡萄栽培在全球葡萄酒行业中具有重要的经济意义。目前,墨西哥种植葡萄的面积约为 28500 公顷,是世界上第 26 大葡萄种植国。鉴于其重要性,早期发现植物病害的病原体对于预防疾病爆发至关重要。因此,我们的研究旨在鉴定表现出树干病害症状的葡萄藤中的真菌菌株,并评估其酶活性作为其植物病原潜力的指标。我们从墨西哥的克雷塔罗州收集了包括马尔贝克、设拉子和丹魄在内的多个葡萄品种。在实验室中,我们轻轻去除植物的树皮,以防止外部污染。然后,对样本进行表面消毒,并从有症状的组织中产生木屑。我们使用无菌技术从回收的木屑中分离可培养的真菌菌株。将菌落接种在 PDA 上,并通过显微镜观察和 ITS 和 LSU nrDNA 区域的 DNA 测序相结合,以及在 GenBank 数据库中进行 BLASTn 搜索来鉴定。我们使用特定的培养基和染色剂评估了菌株降解纤维素、淀粉和木质素的定性能力。通过培养形态和 DNA 测序,确定了七个属中的 13 个种:节菱孢属、曲霉属、枝孢属、离蠕孢属、镰孢属、拟无梗囊霉属和壳球孢属。一些分离的菌株能够降解纤维素或木质素或淀粉。这些结果是这些物种在美洲的首次报道。使用基于培养的和 DNA 测序工具可以继续检测真菌菌株,以进行早期的 GTD 预防。

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