Sustainable Agro-Ecosystems and Bioresources Department, IASMA Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige, Italy.
Appl Environ Microbiol. 2012 Jun;78(12):4308-17. doi: 10.1128/AEM.07655-11. Epub 2012 Apr 6.
We studied the distribution of fungal endophytes of grapevine (Vitis vinifera L.) plants in a subalpine area of northern Italy, where viticulture is of high economic relevance. We adopted both cultivation-based and cultivation-independent approaches to address how various anthropic and nonanthropic factors shape microbial communities. Grapevine stems were harvested from several locations considering organic and integrated pest management (IPM) and from the cultivars Merlot and Chardonnay. Cultivable fungi were isolated and identified by internal-transcribed-spacer sequence analysis, using a novel colony-PCR method, to amplify DNA from fungal specimens. The composition of fungal communities was assessed using a cultivation-independent approach, automated ribosomal intergenic spacer analysis (ARISA). Multivariate statistical analysis of both culture-dependent and culture-independent data sets was convergent and indicated that fungal endophytic communities in grapevines from organically managed farms were different from those from farms utilizing IPM. Fungal communities in plants of cv. Merlot and cv. Chardonnay overlapped when analyzed using culture-dependent approaches but could be partially resolved using ARISA fingerprinting.
我们研究了意大利北部亚高山地区葡萄(Vitis vinifera L.)植物内生真菌的分布,该地区的葡萄种植业具有重要的经济意义。我们采用了基于培养和非培养的方法,以研究各种人为和非人为因素如何塑造微生物群落。从多个考虑有机和综合虫害管理(IPM)的地点以及 Merlot 和 Chardonnay 品种收获葡萄藤茎。使用新型的菌落-PCR 方法从真菌标本中扩增 DNA,通过内部转录间隔区序列分析对可培养真菌进行分离和鉴定。使用非培养方法自动核糖体基因间 spacer 分析(ARISA)评估真菌群落的组成。基于培养和非培养数据集的多元统计分析结果一致,表明有机管理农场的葡萄内生真菌群落与采用 IPM 的农场的内生真菌群落不同。使用基于培养的方法分析 Merlot 和 Chardonnay 品种的植物时,真菌群落重叠,但使用 ARISA 指纹图谱可部分解决。