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MicroRNAs in the exosome-like nanoparticles from orange juice inhibit Citrus blue mold caused by Penicillium italicum [miRNA-Seq]

ABSTRACT: Identification of miRNAs in citrus reticulata exosomes；Identification of potential target genes of exosomal miRNAs in penicillium italicum； Comparison of differentially expressed genes between citrus exosome-treatedpenicillium italicum and wild type

ORGANISM(S): Citrus reticulata

PROVIDER: GSE214036 | GEO | 2022/09/26

REPOSITORIES: GEO

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MicroRNAs in the exosome-like nanoparticles from orange juice inhibit Citrus blue mold caused by Penicillium italicum [degradome]

Project description:Identification of miRNAs in citrus reticulata exosomes；Identification of potential target genes of exosomal miRNAs in penicillium italicum； Comparison of differentially expressed genes between citrus exosome-treatedpenicillium italicum and wild type

2022-09-26 | GSE214037 | GEO

MicroRNAs in the exosome-like nanoparticles from orange juice inhibit Citrus blue mold caused by Penicillium italicum [RNA-Seq]

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MicroRNAs in the exosome-like nanoparticles from orange juice inhibit Citrus blue mold caused by Penicillium italicum

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Silencing Dicer-Like Genes Reduces Virulence and sRNA Generation in Penicillium italicum, the Cause of Citrus Blue Mold.

Project description:The Dicer protein is one of the most important components of RNAi machinery because it regulates the production of small RNAs (sRNAs) in eukaryotes. Here, Dicer1-like gene (Pit-DCL1) and Dicer2-like gene (Pit-DCL2) RNAi transformants were generated via pSilent-1 in Penicillium italicum (Pit), which is the causal agent of citrus blue mold. Neither transformant showed a change in mycelial growth or sporulation ability, but the pathogenicity of the Pit-DCL2 RNAi transformant to citrus fruits was severely impaired, compared to that of the Pit-DCL1 RNAi transformant and the wild type. We further developed a citrus wound-mediated RNAi approach with a double-stranded fragment of Pit-DCL2 generated in vitro, which achieved an efficiency in reducing Pi-Dcl2 expression and virulence that was similar to that of protoplast-mediated RNAi in P. italicum, suggesting that this approach is promising in the exogenous application of dsRNA to control pathogens on the surface of citrus fruits. In addition, sRNA sequencing revealed a total of 69.88 million potential sRNAs and 12 novel microRNA-like small RNAs (milRNAs), four of which have been predicated on target innate immunity or biotic stress-related genes in Valencia orange. These data suggest that both the Pit-DCL1 and Pit-DCL2 RNAi transformants severely disrupted the biogenesis of the potential milRNAs, which was further confirmed for some milRNAs by qRT-PCR or Northern blot analysis. These data suggest the sRNAs in P. italicum that may be involved in a molecular virulence mechanism termed cross-kingdom RNAi (ck-RNAi) by trafficking sRNA from P. italicum to citrus fruits.

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Deep sequencing discovery of novel and conserved microRNAs in trifoliate orange (Citrus trifoliata)

Project description:MicroRNAs (miRNAs) play a critical role in post-transcriptional gene regulation. miRNAs have been shown to control many genes involved in various biological and metabolic processes. Deep sequencing technologies have facilitated identification of species-specific or lowly expressed as well as conserved or highly expressed miRNAs in plants. In this research, we used Solexa sequencing to discover new microRNAs in trifoliate orange (Citrus trifoliata) an important rootstock of citrus. A total of 13,106,753 reads representing 4,876,395 distinct sequences were obtained from a short RNA library generated from small RNA extracted from C. trifoliata flower and fruit tissues, Based on sequence similarity and hairpin structure prediction, we found that 178,102 reads representing 89 sequences from 42 highly conserved miRNA families, have perfect matches to known miRNAs. We also identified 10 novel miRNA candidates, whose precursors were all potentially generated from citrus ESTs. And of them five miRNA* sequences were also sequenced. These sequences had not been described in other plant species and accumulation of these 10 novel miRNAs were confirmed by qRT-PCR analysis. Potential target genes were predicted for most conserved and novel miRNAs. Moreover, four target genes included one encoding IRX12 copper ion binding/ oxidoreductase and three genes encoding NB-LRR disease resistance protein have been experimentally verified by detection of the miRNA-mediated mRNA cleavage in C. trifoliata.

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