Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi di Bari Aldo Moro, Via Amendola 165/A, 70126 Bari, Italy.
Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi di Bari Aldo Moro, Via Amendola 165/A, 70126 Bari, Italy.
Int J Food Microbiol. 2020 Feb 2;314:108389. doi: 10.1016/j.ijfoodmicro.2019.108389. Epub 2019 Oct 22.
Most of diseases of pomegranate fruit are caused by fungal pathogens, which provoke postharvest yield and economical losses. Aspergillus and Penicillium sensu lato (s.l.) are the main wound pathogens of pomegranate fruit. In the present investigation, the populations of Aspergillus and Penicillium s.l. isolated from pomegranate fruit in Southern Italy were characterized. Since the morphological identification of species belonging to these genera is laborious, molecular approaches, such as PCR and High-Resolution Melting (HRM), were used. Particularly, a specific primer pair was designed to discriminate, within the Penicillium s.l. population, Penicillium sensu stricto (s.s.) from Talaromyces strains. Then, a new HRM assay for species identification within Penicillium s.s. according to SNPs present in a portion of the beta-tubulin gene was set up. Similarly, Aspergillus sect. nigri population was characterized arranging a HRM assay, whose primer pair was designed on a portion of the calmodulin gene. According to these assays, 10% of the Penicillium s.l. population proved to be made up of Talaromyces biverticillius strains. Furthermore, six species of Penicillium s.s. (P. adametzioides, P. brevicompactum, P. citrinum, P. glabrum, P. pagulum, and P. johnkrugii) and four of black aspergilli (A. tubingensis, A. welwitschiae, A. japonicus, and A. uvarum) were identified; all species belonging to both genera disclosed different incidences in postharvest rotted pomegranate fruit. Moreover, since Aspergillus and Penicillium are potentially producers of mycotoxins, like ochratoxin A and fumonisins, the presence/absence of genes involved in mycotoxin biosynthetic pathways was tested. Some Aspergillus strains belonging to species A. welwitschiae proved to possess fumonisin genes. The setup of molecular tools to characterize Penicillium s.l. and Aspergillus sect. nigri species infecting pomegranate fruit after harvest is of paramount importance for their effective control, even more considering the ability of these fungal genera to produce mycotoxins, which are hazardous for human health and potentially present also in by-products.
石榴果实的大多数疾病是由真菌病原体引起的,这些病原体导致了石榴果实的产后产量和经济损失。曲霉属和 Penicillium sensu lato(s.l.)是石榴果实的主要伤口病原体。在本研究中,对来自意大利南部石榴果实中的曲霉属和 Penicillium s.l. 种群进行了特征描述。由于这些属的物种形态鉴定非常繁琐,因此使用了分子方法,如 PCR 和高分辨率熔解(HRM)。特别是,设计了一对特定的引物来区分 Penicillium s.l. 种群中的 Penicillium sensu stricto(s.s.)和 Talaromyces 菌株。然后,根据β-微管蛋白基因的一部分中存在的 SNP,建立了一种用于鉴定 Penicillium s.s. 内种的新 HRM 检测方法。同样,通过设计一组钙调蛋白基因的引物,对黑曲霉属种群进行了特征描述。根据这些检测方法,10%的 Penicillium s.l. 种群被证明由 Talaromyces biverticillius 菌株组成。此外,还鉴定出了六个 Penicillium s.s.(P. adametzioides、P. brevicompactum、P. citrinum、P. glabrum、P. pagulum 和 P. johnkrugii)和四个黑曲霉(A. tubingensis、A. welwitschiae、A. japonicus 和 A. uvarum);这两个属的所有物种在产后腐烂的石榴果实中都有不同的发病率。此外,由于曲霉属和青霉属是赭曲霉毒素 A 和伏马菌素等真菌毒素的潜在生产者,因此测试了参与真菌毒素生物合成途径的基因的存在/缺失情况。一些属于 A. welwitschiae 种的曲霉属菌株被证明拥有伏马菌素基因。建立用于描述产后感染石榴果实的 Penicillium s.l. 和 Aspergillus sect. nigri 物种的分子工具对于它们的有效控制至关重要,特别是考虑到这些真菌属产生真菌毒素的能力,这些毒素对人类健康有害,并且也可能存在于副产品中。