Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto Río Cuarto, Córdoba, Argentina.
Braz J Microbiol. 2013 Oct 30;44(2):447-55. doi: 10.1590/S1517-83822013000200017. eCollection 2013.
The objectives of this study were to evaluate the ability to produce alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TA) by A. alternata and A. infectoria strains recovered from wheat kernels obtained from one of the main production area in Argentina; to confirm using AFLPs molecular markers the identify of the isolates up to species level, and to evaluate the intra and inter-specific genetic diversity of these two Alternaria species. Among all the Alternaria strains tested (254), 84% of them were able to produce mycotoxins. The most frequent profile of toxin production found was the co-production of AOH and AME in both species tested. TA was only produced by strains of A. alternata. Amplified fragment polymorphism (AFLPs) analysis was applied to a set of 89 isolates of Alternaria spp (40 were A. infectoria and 49 were A. alternata) in order to confirm the morphological identification. The results showed that AFLPs are powerful diagnostic tool for differentiating between A. alternata and A. infectoria. Indeed, in the current study the outgroup strains, A. tenuissima was consistently classified. Characteristic polymorphic bands separated these two species regardless of the primer combination used. Related to intraspecific variability, A. alternata and A. infectoria isolates evaluated seemed to form and homogeneous group with a high degree of similarity among the isolates within each species. However, there was more scoreable polymorphism within A. alternata than within A. infectoria isolates. There was a concordance between morphological identification and separation up to species level using molecular markers. Clear polymorphism both within and between species showed that AFLP can be used to asses genetic variation in A. alternata and A. infectoria. The most important finding of the present study was the report on AOH and AME production by A. infectoria strains isolated from wheat kernels in Argentina on a semisynthetic media for the first time. Also, specific bands for A. alternata and A. infectoria have been identified; these may be useful for the design of specific PCR primers in order to differentiate these species and to detect them in cereals.
本研究的目的是评估从阿根廷主要生产区之一收获的小麦籽粒中回收的交替镰刀菌(A. alternata)和黄萎镰刀菌(A. infectoria)菌株产生 alternariol(AOH)、alternariol monomethyl ether(AME)和 tenuazonic acid(TA)的能力;使用 AFLP 分子标记确认分离株的种水平鉴定,并评估这两种 Alternaria 物种的种内和种间遗传多样性。在所测试的所有交替镰刀菌菌株(254 株)中,84%能够产生真菌毒素。在所测试的两种物种中,发现最常见的产毒谱是 AOH 和 AME 的共同产生。TA 仅由 A. alternata 菌株产生。扩增片段多态性(AFLPs)分析应用于一组 89 株 Alternaria spp 分离株(40 株为 A. infectoria,49 株为 A. alternata),以确认形态学鉴定。结果表明,AFLPs 是区分 A. alternata 和 A. infectoria 的有效诊断工具。事实上,在本研究中,外群菌株 A. tenuissima 始终被分类。无论使用哪种引物组合,特征多态性条带均可将这两个物种分开。关于种内变异性,评估的 A. alternata 和 A. infectoria 分离株似乎形成了一个同质组,每个种内的分离株之间具有高度相似性。然而,A. alternata 分离株内的可评分多态性高于 A. infectoria 分离株内。形态学鉴定与使用分子标记进行种水平分离之间存在一致性。种内和种间的明显多态性表明,AFLP 可用于评估 A. alternata 和 A. infectoria 的遗传变异。本研究的一个重要发现是首次在半合成培养基上从阿根廷小麦籽粒中分离出的 A. infectoria 菌株产生 AOH 和 AME。此外,还鉴定了 A. alternata 和 A. infectoria 的特异性条带;这些可能有助于设计特定的 PCR 引物,以区分这些物种并在谷物中检测它们。