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Screening and Identification of Novel Ochratoxin A-Producing Fungi From Grapes

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Screening and Identification of Novel Ochratoxin A-Producing Fungi From Grapes

Xiaoyun Zhang et al. Toxins (Basel).

Abstract

Ochratoxin A (OTA) contamination has been established as a world-wide problem. In this study, the strains with the ability of OTA production were screened by analyzing the green fluorescence of the isolates colonies from the grapes in Zhenjiang with 365 nm UV light and confirmed by HPLC with fluorescent detection (HPLC-FLD). The results showed that seven isolates acquired the characteristic of the fluorescence, of which only five showed the ability of OTA production as confirmed by HPLC-FLD analysis. The five OTA-producing strains were identified based on comparative sequence analysis of three conserved genes (ITS, BenA and RPB2) of the strains, and they are Talaromyces rugulosus (O1 and Q3), Penicillium commune (V5-1), Penicillium rubens (MQ-5) and Aspergillus aculeatus (MB1-1). There are two Penicillium species of the five OTA-producing strains and our study is the first to report that P. rubens, T. rugulosus and A. aculeatus can produce OTA. This work would contribute to comprehensively understanding the fungi with an OTA-producing ability in grapes before harvest and then take effective measures to prevent OTA production.

Keywords: fungi; grapes; identification; ochratoxin A (OTA); screening.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Green fluorescence (a) on the reverse of the colony of O1 (an Ochratoxin A (OTA)-producing strain) grown on Czapek’s Agar medium (CA) when was exposed to long wavelength UV light (365 nm). No fluorescence (b) was displayed by a non-OTA-producing strain in the same conditions.
Figure 2
Figure 2
The phylogenetic tree of the O1 strain by the Neighbor-Joining method based on analysis of ITS, BenA and RPB2 sequences. (a) the phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.
Figure 2
Figure 2
The phylogenetic tree of the O1 strain by the Neighbor-Joining method based on analysis of ITS, BenA and RPB2 sequences. (a) the phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.
Figure 3
Figure 3
The phylogenetic tree of the Q3 strain by the Neighbor-Joining method based on analysis of ITS, BenA and RPB2 sequences. (a) the phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.
Figure 4
Figure 4
The phylogenetic tree of the V5-1 strain by the Neighbor-Joining method based on analysis of ITS, BenA. and RPB2 sequences. (a) The phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.
Figure 5
Figure 5
The phylogenetic tree of the MQ5 strain by the Neighbor-Joining method based on analysis of ITS, BenA and RPB2 sequences. (a) the phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.
Figure 6
Figure 6
The phylogenetic tree of the MB1-1 strain by the Neighbor-Joining method based on analysis of ITS, BenA and RPB2 sequences. (a) The phylogenetic tree based on analysis of ITS; (b) the phylogenetic tree based on analysis of BenA; and (c) the phylogenetic tree based on analysis of RPB2.

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