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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2015, Vol. 41 Issue (5): 567-576    DOI: 10.3785/j.issn.1008-9209.2015.03.061
食品科学     
花生中巨大芽孢杆菌对黄曲霉毒素合成相关基因的抑制
孔青, 迟晨, 单世华, 李琦玉
1.中国海洋大学食品科学与工程学院，山东 青岛266003；2.山东省花生研究所，山东 青岛 266100
Inhibitory effect of Bacillus megaterium on aflatoxin biosynthetic pathway genes expression of Aspergillus flavus in peanuts
Kong Qing, Chi Chen, Shan Shihua, Li Qiyu
(1. School of Food Science and Engineering, Ocean University of China, Qingdao 266003, Shandong, China; 2. Shandong Peanut Institute, Qingdao 266100, Shandong, China)
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摘要： 分别在UF 715133-1和Jinhua 1012花生仁上接种黄曲霉和巨大芽孢杆菌,高压液相色谱法测定花生中黄曲霉毒素含量,借助生物芯片和实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qRT-PCR)测定巨大芽孢杆菌对黄曲霉毒素生物合成途径基因表达的影响,并用基因本体(gene ontology, GO)功能分类和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路方法对基因进行分类分析。在实验组Jinhua 1012 和UF 715133-1中,黄曲霉毒素B1的含量降低了85%以上,一些编码蛋白酶的基因显著下调,如短链脱氢酶、非核糖体多肽合成酶等。结果表明,从海洋中分离出的巨大芽孢杆菌可显著抑制黄曲霉毒素在花生上的合成,黄曲霉中多种基因的表达受到抑制,但这些基因在黄曲霉毒素生物合成中的作用并未阐明,其作用机制还有待进一步深入研究。
Abstract: Previous studies in our laboratory demonstrated that a marine Bacillus megaterium could inhibit the growth of Aspergillus flavus and turn off the biosynthesis of aflatoxin B1. In order to investigate the mechanisms of inhibition by B. megaterium on A. flavus growth and aflatoxin production in peanuts and to evaluate the possible application of B. megaterium as a biocontrol agent, genes expression analysis using whole A. flavus genome gene chip when A. flavus was co-cultured with B. megaterium in peanuts were performed. The aim of this research was to study the inhibitory effect of B. megaterium on aflatoxin biosynthetic pathway genes expression in A. flavus. The effects of B. megaterium on aflatoxin biosynthesis and genes expression in A. flavus were tested in two types of peanut varieties, UF 715133-1 and Jinhua 1012 by the aid of high performance liquid chromatography (HPLC), gene chip and quantitative real-time polymerase chain reaction (qRT-PCR). UF 715133-1 is a kind of peanut that could resist to A. flavus invasion, while Jinhua 1012 is susceptible to the infection of A. flavus. The results showed that though more than 85% of aflatoxin B1 was inhibited by the treatment of B. megaterium in UF 715133-1 and Jinhua 1012, no important aflatoxin biosynthetic pathway gene was differentially expressed. Peanuts  unsuitable for the growth of B. megaterium was the main reason for B. megaterium  unthoroughly inhibit the biosynthesis of aflatoxin like in minimal medium (MM) or potato dextrose broth (PDB). Furthermore, the experimental condition (28 ℃, relative humidity 85%), which is the optimal condition for aflatoxin biosynthesis, wasn’t suitable for the growth of B. megaterium neither. So we inferred that B. megaterium inhibited aflatoxin biosynthesis by inhibiting the growth of A. flavus in peanuts, and the secondary metabolites in B. megaterium accounted for the inhibition. Furthermore, the results showed some coding protease genes were highly downregulated, such as short-chain dehydrogenase, and non-ribosomal peptide synthetase,  which may relate to aflatoxin biosynthesis. In conclusion, this strain of marine B. megaterium isolated from the Yellow Sea of East China could significantly inhibit the aflatoxin biosynthesis in peanuts through inhibiting genes expression in A. flavus. While the role of most highly downregulated genes in aflatoxin biosynthesis hasn’t been clarified, the mechanism remains to be further studied.
出版日期: 2015-09-20
CLC:  S 565.2  
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孔青
迟晨
单世华
李琦玉

引用本文:

孔青, 迟晨, 单世华, 李琦玉. 花生中巨大芽孢杆菌对黄曲霉毒素合成相关基因的抑制[J]. 浙江大学学报(农业与生命科学版), 2015, 41(5): 567-576.

Kong Qing, Chi Chen, Shan Shihua, Li Qiyu. Inhibitory effect of Bacillus megaterium on aflatoxin biosynthetic pathway genes expression of Aspergillus flavus in peanuts. Journal of Zhejiang University (Agriculture and Life Sciences), 2015, 41(5): 567-576.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2015.03.061    或    http://www.zjujournals.com/agr/CN/Y2015/V41/I5/567

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