The Application of siRNA Interference Technique in Aflatoxin Inhibition

doi:10.11924/j.issn.1000-6850.casb18100089

Abstract

Abstract:

Traditional methods for constructing fungal gene knockout mutants would take a long experimental period. In order to rapidly detect and high-throughput screen target genes with specific functions, small molecule RNA interference technique was used to silence the key genes of aflatoxin biosynthesis in this study, which effectively reduced the biosynthesis of aflatoxin B₁ (AFB₁). By designing and synthesizing artificial small interfering RNAs (artificial siRNAs) which specifically targeted the key gene aflR in aflatoxin synthesis pathway, artificial siRNAs were directly transformed into the protoplast of Aspergillus flavus, which effectively silenced the key regulator aflR gene of aflatoxin synthesis, and led to significantly decreasing AFB₁ biosynthesis. The siRNAs of the other 11 target genes in the aflatoxin synthesis pathway were used in this study, protoplast transformation could directly silence these genes, and the transcription levels of these genes were significantly reduced, and among them, the amount of AFB₁ biosynthesis decreased significantly after silencing hypC and aflW gene, compared to the control. In addition, using siRNA specifically targeting aflatoxin synthesize gene clusters, such as the pks-nrps gene, silencing of pks-nrps gene by siRNA interference could also significantly reduce the synthesis of AFB₁. Therefore, this study relies on small molecule RNA interference technique and established a reverse genetics method for rapid detection and high-throughput screening of aflatoxin biosynthesis pathway-related genes.

Key words: small RNA, gene silencing, aflatoxin, RNA interference, reverse genetics technology

CLC Number:

Jiao Jian, Du Peng. The Application of siRNA Interference Technique in Aflatoxin Inhibition[J]. Chinese Agricultural Science Bulletin, 2020, 36(5): 30-36.

Figures/Tables 9

References 22

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