RNA Silencing of Mycotoxin Production in Aspergillus and Fusarium Species

McDonald, Tami; Brown, Daren W.; Keller, Nancy P.; Hammond, Thomas M.

doi:10.1094/mpmi-18-0539

Cited by 111 publications

(81 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Localization of BioF to the Peroxisome and Complementation of the ⌬bioF Mutant-The GFP-bioF fragments were excised from plasmids pTRC/GFP-BioF-FL and pTRC/GFP-BioF-TR (see above) with NcoI-BamHI and cloned into the same restriction sites of plasmid pTMH44.2 (21), which placed the fusion proteins under the control of the A. nidulans gpdA promoter and trpC terminator. The promoter-GFP-bioF-terminator expression cassette was excised by EcoRI-XbaI and cloned into the EcoRI-SpeI restriction sites of vector pMT1612, which contains the bar gene from Streptomyces hygroscopicus, conferring resistance to the herbicide glufosinate (DL-phosphinothricin) (19).…”

Section: Complementation Of E Coli Biof-deficient Mutants With Gfp-bmentioning

confidence: 99%

Contributions of the Peroxisome and β-Oxidation Cycle to Biotin Synthesis in Fungi

Magliano

Flipphi

Arpat

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Pimeloyl-CoA is an intermediate of the biotin pathway, but it is unknown how it is synthesized in fungi. Results: Aspergillus nidulans mutants in ␤-oxidation are biotin-deficient, and the enzyme utilizing pimeloyl-CoA is targeted to the peroxisome. Conclusion: Pimeloyl-CoA is synthesized in the peroxisome via the ␤-oxidation cycle in filamentous fungi. Significance: This is the first implication of peroxisomal ␤-oxidation in biotin synthesis.

show abstract

Section: Complementation Of E Coli Biof-deficient Mutants With Gfp-bmentioning

confidence: 99%

Contributions of the Peroxisome and β-Oxidation Cycle to Biotin Synthesis in Fungi

Magliano

Flipphi

Arpat

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Recently, RNA interference (RNAi) has emerged as a powerful tool for gene targeting in fungi [26][27][28]. Conventional RNAi machinery components, including the RNA-dependent RNA polymerase, Argonaut proteins and Dicers are conserved in some fungi, e.g., Cryptococcus neoformans [29], Bipolaris oryzae [30], Aspergillus and Fusarium Species [31] and Cochliobolus sativus [32]. However, its application in C. globosum has not been reported.…”

mentioning

confidence: 99%

A PKS gene, pks-1, is involved in chaetoglobosin biosynthesis, pigmentation and sporulation in Chaetomium globosum

Hao

Lou

et al. 2012

Sci. China Life Sci.

View full text Add to dashboard Cite

Chaetomium globosum is one of the most common fungi in nature. It is best known for producing chaetoglobosins; however, the molecular basis of chaetoglobosin biosynthesis is poorly understood in this fungus. In this study, we utilized RNA interference (RNAi) to characterize a polyketide synthase gene, pks-1, in C. globosum that is involved in the production of chaetoglobosin A. When pks-1 was knocked down by RNAi, the production of chaetoglobosin A dramatically decreased. Knock-down mutants also displayed a pigment-deficient phenotype. These results suggest that the two polyketides, melanin and chaetoglobosin, are likely to share common biosynthetic steps. Most importantly, we found that pks-1 also plays a critical role in sporulation. The silenced mutants of pks-1 lost the ability to produce spores. We propose that polyketides may modulate cellular development via an unidentified action. We also suggest that C. globosum pks-1 is unique because of its triple role in melanin formation, chaetoglobosin biosynthesis and sporulation. This work may shed light on chaetoglobosin biosynthesis and indicates a relationship between secondary metabolism and fungal morphogenesis.

show abstract

“…Thus, aflR was one of the targets for silencing in vector p5XCAPD, but was not the only one. Inverted repeats of the aflR gene introduced into A. flavus and A. parasiticus by transformation resulted in silencing and minimal or no production of aflatoxins 47 ( McDonald et al, 2005b). Also, silencing aflD gene prevented aflatoxin production by up to 98% in A. flavus and A. parasiticus in direct transformation 48 .…”

Section: Discussionmentioning

confidence: 99%

RNAi-mediated Control of Aflatoxins in Peanut: Method to Analyze Mycotoxin Production and Transgene Expression in the Peanut/Aspergillus Pathosystem

Arias

Dang

Соболев

2015

JoVE

View full text Add to dashboard Cite

Citation: Arias, R.S., Dang, P.M., Sobolev, V.S. RNAi-mediated Control of Aflatoxins in Peanut: Method to Analyze Mycotoxin Production and Transgene Expression in the Peanut/Aspergillus Pathosystem. J. Vis. Exp. (106), e53398, doi:10.3791/53398 (2015). AbstractThe Food and Agriculture Organization of the United Nations estimates that 25% of the food crops in the world are contaminated with aflatoxins. That represents 100 million tons of food being destroyed or diverted to non-human consumption each year. Aflatoxins are powerful carcinogens normally accumulated by the fungi Aspergillus flavus and A. parasiticus in cereals, nuts, root crops and other agricultural products. Silencing of five aflatoxin-synthesis genes by RNA interference (RNAi) in peanut plants was used to control aflatoxin accumulation following inoculation with A. flavus. Previously, no method existed to analyze the effectiveness of RNAi in individual peanut transgenic events, as these usually produce few seeds, and traditional methods of large field experiments under aflatoxin-conducive conditions were not an option. In the field, the probability of finding naturally contaminated seeds is often 1/100 to 1/1,000. In addition, aflatoxin contamination is not uniformly distributed. Our method uses few seeds per transgenic event, with small pieces processed for real-time PCR (RT-PCR) or small RNA sequencing, and for analysis of aflatoxin accumulation by ultra-performance liquid chromatography (UPLC). RNAi-expressing peanut lines 288-72 and 288-74, showed up to 100% reduction (p≤0.01) in aflatoxin B 1 and B 2 compared to the control that accumulated up to 14,000 ng . This protocol describes the application of RNAi-mediated control of aflatoxins in transgenic peanut seeds and methods for its evaluation. We believe that its application in breeding of peanut and other crops will bring rapid advancement in this important area of science, medicine and human nutrition, and will significantly contribute to the international effort to control aflatoxins, and potentially other mycotoxins in major food crops.

show abstract

RNA Silencing of Mycotoxin Production in Aspergillus and Fusarium Species

Cited by 111 publications

References 32 publications

Contributions of the Peroxisome and β-Oxidation Cycle to Biotin Synthesis in Fungi

Contributions of the Peroxisome and β-Oxidation Cycle to Biotin Synthesis in Fungi

A PKS gene, pks-1, is involved in chaetoglobosin biosynthesis, pigmentation and sporulation in Chaetomium globosum

RNAi-mediated Control of Aflatoxins in Peanut: Method to Analyze Mycotoxin Production and Transgene Expression in the Peanut/<em>Aspergillus</em> Pathosystem

Contact Info

Product

Resources

About