广陈皮外源真菌的组成及产毒真菌分析

Toxin-producing fungi and exogenous fungi on Citri reticulatae pericarpium

摘要:

【背景】广陈皮为药食同源中药材，在高温、高湿且贮存不当的条件下容易发霉，从而产生毒素，严重威胁陈皮的质量安全。【目的】分析广陈皮表面外源真菌的组成及其产生毒素的真菌。【方法】采用平板稀释法分离广陈皮表面外源真菌，利用分生孢子形态特征及DNA序列分析进行真菌鉴定，采用高效液相色谱-三重串联四极杆质谱联用技术对青霉属和曲霉属进行产毒检测。【结果】从广陈皮表面分离外源真菌共132株，鉴定为子囊菌亚门(Ascomycota，98.48%)和毛霉菌亚门(Mucoromycota，1.52%)，包括散囊菌纲(Eurotiomycetes，95.45%)、座囊菌纲(Dothideomycetes，3.03%)和毛霉纲(Mucoromycetes，1.52%)。分离活动曲霉属(Aspergillus spp.)菌株共77株，为广陈皮药材外源真菌的优势菌，青霉属(Penicillium spp.)次之。毒素检测筛选出1株产毒真菌，鉴定为黄曲霉(Aspergillusflavus) JXCP1-3。该菌株产生黄曲霉毒素B₁ (aflatoxin B₁，AFB₁)，浓度为1.52 ng/mL。【结论】全面分析广陈皮表面的外源真菌及产毒真菌，为广陈皮表面产毒真菌的防控提供依据。

Abstract:

[Background] Citri reticulatae pericarpium is a widely used Chinese medicinal material with both medical and edible values.The growing of fungi and the production of mycotoxins by fungi during storage at proper humidity and temperature pose a threat to the quality and safety of Citri reticulatae pericarpium.[Objective] In order to analyze the composition of exogenous fungi and the toxin-producing fungi on Citri reticulatae pericarpium.[Methods] The exogenous fungi were isolated by plate dilution method and identified based on the morphological characteristics and sequence analysis.The toxin-producing fungi were detected by ultra-high performance liquid chromatography coupled to tandem quadrupole mass spectrometry (UPLC-MS/MS).[Results] A total of 132 strains of exogenous fungi were isolated from Citri reticulatae pericarpium.They were identified to belong to Ascomycota(98.48%) and Mucoromycota(1.52%),including Eurotiomycetes(95.45%),Dothideomycetes(3.03%) and Mucoromycetes(1.52%).Seventy-seven strains belonged to Aspergillus spp.,which was dominant on Citri reticulatae pericarpium,followed by Penicillium spp.The strains of Aspergillus spp.and Penicillium spp.were detected by UPLC-MS/MS.One strain of toxin-producing fungi was screened out and identified as A.flavus JXCP1-3,which can produce 1.52 ng/mL aflatoxin B₁.[Conclusion] We comprehensively analyzed the exogenous fungi and toxin-producing fungi on the surface of Citri reticulatae pericarpium,which provided a basis for the prevention and control of the toxin-producing fungi.