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金属离子调节霉菌毒素黄曲霉毒素和玉米赤霉烯酮的基因表达及积累。

Metal ions modulate gene expression and accumulation of the mycotoxins aflatoxin and zearalenone.

作者信息

Cuero R, Ouellet T

机构信息

Prairie View A & M University, CARC, Texas A & M University System, Prairie View, TX 77446, USA.

出版信息

J Appl Microbiol. 2005;98(3):598-605. doi: 10.1111/j.1365-2672.2004.02492.x.

Abstract

AIMS

To determine the modulating action of some metal ions (Zn+2, Fe+2, Cu+2) on gene expression of enzymes related to fungal growth and accumulation of the mycotoxins aflatoxin and zearalenone.

METHODS AND RESULTS

The effect of the metal ions, as single or mixed treatments, was observed in submerged cultures of toxigenic Aspergillus flavus or Fusarium graminearum, which produce the mycotoxins aflatoxin or zearalenone, respectively. The enzyme-linked immunosorbent assay results showed that the single metals Zn+2 or Cu+2 stimulated aflatoxin accumulation while Cu+2 or Fe+2 stimulated zearalenone in fungal cultures. Single Zn+2 treatment also affected conidial differentiation and pigmentation. A cDNA suppression subtractive library was also produced and followed by sequencing of potential metal treatment-specific clones, thus determining induced genes. The genes uncovered included enzymes and regulators of cell growth and division, including many genes with unknown functions were uncovered. A Northern blot analysis was used to verify the expression pattern of the corresponding genes under metal treatment. The metal ions enhanced the expression of alcohol dehydrogenase Adh1 homologue by up to 33-fold in A. flavus and ca fourfold in F. graminearum. Encoding homologues of a neutral amino acid permease, were also used in the Northern analysis. However, the expression of the permease was not significantly affected by metal ion treatments.

CONCLUSIONS

The results showed a significant effect of metal ions on expression of gene related to fungal growth, development, conidiation and production of both aflatoxin and zearalenone.

SIGNIFICANT AND IMPACT OF THE STUDY

At the molecular and cellular level, the significant effects of metal ions on fungal growth and development, conidiation, and production of both aflatoxin and zearalenone were demonstrated.

摘要

目的

确定某些金属离子(Zn²⁺、Fe²⁺、Cu²⁺)对与真菌生长以及霉菌毒素黄曲霉毒素和玉米赤霉烯酮积累相关的酶的基因表达的调节作用。

方法与结果

在分别产生霉菌毒素黄曲霉毒素或玉米赤霉烯酮的产毒黄曲霉或禾谷镰刀菌的液体培养物中,观察了金属离子作为单一处理或混合处理的效果。酶联免疫吸附测定结果表明,单一金属Zn²⁺或Cu²⁺刺激了黄曲霉毒素的积累,而Cu²⁺或Fe²⁺刺激了真菌培养物中玉米赤霉烯酮的产生。单一Zn²⁺处理还影响了分生孢子分化和色素沉着。还构建了一个cDNA抑制消减文库,随后对潜在的金属处理特异性克隆进行测序,从而确定诱导基因。发现的基因包括细胞生长和分裂的酶和调节因子,其中包括许多功能未知的基因。使用Northern印迹分析来验证金属处理下相应基因的表达模式。金属离子使黄曲霉中乙醇脱氢酶Adh1同源物的表达增强了高达33倍,在禾谷镰刀菌中增强了约4倍。编码中性氨基酸通透酶同源物的基因也用于Northern分析。然而,通透酶的表达不受金属离子处理的显著影响。

结论

结果表明金属离子对与真菌生长、发育、分生孢子形成以及黄曲霉毒素和玉米赤霉烯酮产生相关的基因表达有显著影响。

研究的意义和影响

在分子和细胞水平上,证明了金属离子对真菌生长、发育、分生孢子形成以及黄曲霉毒素和玉米赤霉烯酮产生具有显著影响。

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