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Nonfunctional ingestion of plant miRNAs in silkworm revealed by digital droplet PCR and transcriptome analysis.

Jia L, Zhang D, Xiang Z, He N - Sci Rep (2015)

Bottom Line: The results of TA clone, Sanger sequencing and droplet digital PCR demonstrated that several mulberry-derived miRNAs could enter to silkworm hemolymph and multiple tested tissues.Mulberry miRNAs are convincingly transferred to the silkworm orally and no physiological process associated with the miRNAs was demonstrable.The results provided a new aspect of cross-kingdom miRNA transfer.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing 400715, P. R. China.

ABSTRACT
Since a plant miRNA (miR168) cross-regulating a mammalian transcript was reported, miRNA-mediated cross-kingdom communication has become one of the most compelling but controversial topics. In the present study, we used silkworm and mulberry, which is a model for studies on the interactions between the insect and its host plant, to address whether miRNA-mediated cross-kingdom communication is a common phenomenon. The results of TA clone, Sanger sequencing and droplet digital PCR demonstrated that several mulberry-derived miRNAs could enter to silkworm hemolymph and multiple tested tissues. Synthetic miR166b was also detected in hemolymph and fat body. However, the ingestion of synthetic miR166b did not play roles in silkworm physiological progress, which was revealed by RNA-seq analyses, RT-PCR, and phenotypic investigations. Mulberry miRNAs are convincingly transferred to the silkworm orally and no physiological process associated with the miRNAs was demonstrable. The results provided a new aspect of cross-kingdom miRNA transfer.

No MeSH data available.


The expression level of 11 potential miR166b target genes in whole body before and after silkworm ingested synthetic miR166b.FED-control represents the silkworm larvae were fed on only mulberry leaves. FED-miR166b means the silkworm larvae were fed on a piece of mulberry leaf containing synthetic miR166b. Statistical significance was determined by Student’s t test (*p < 0.05).
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f2: The expression level of 11 potential miR166b target genes in whole body before and after silkworm ingested synthetic miR166b.FED-control represents the silkworm larvae were fed on only mulberry leaves. FED-miR166b means the silkworm larvae were fed on a piece of mulberry leaf containing synthetic miR166b. Statistical significance was determined by Student’s t test (*p < 0.05).

Mentions: To explore the functions of miR166b in silkworm, we predicted the potential target genes for miR166b in silkworm full-length cDNA data using miranda software. With the criteria of energy lower than −25 kcal/mol and perfected “seed region” match, we identified 72 potential target genes of miR166b (data not shown). Eleven best matched target genes, whose energy were lower than −26.84 kcal/mol (supplementary Table S3) were subjected to RT-PCR verification. The results showed the expression levels of these genes were not significantly changed in silkworm fed on synthetic miR166b (Fig. 2, supplementary Fig. S2).


Nonfunctional ingestion of plant miRNAs in silkworm revealed by digital droplet PCR and transcriptome analysis.

Jia L, Zhang D, Xiang Z, He N - Sci Rep (2015)

The expression level of 11 potential miR166b target genes in whole body before and after silkworm ingested synthetic miR166b.FED-control represents the silkworm larvae were fed on only mulberry leaves. FED-miR166b means the silkworm larvae were fed on a piece of mulberry leaf containing synthetic miR166b. Statistical significance was determined by Student’s t test (*p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508662&req=5

f2: The expression level of 11 potential miR166b target genes in whole body before and after silkworm ingested synthetic miR166b.FED-control represents the silkworm larvae were fed on only mulberry leaves. FED-miR166b means the silkworm larvae were fed on a piece of mulberry leaf containing synthetic miR166b. Statistical significance was determined by Student’s t test (*p < 0.05).
Mentions: To explore the functions of miR166b in silkworm, we predicted the potential target genes for miR166b in silkworm full-length cDNA data using miranda software. With the criteria of energy lower than −25 kcal/mol and perfected “seed region” match, we identified 72 potential target genes of miR166b (data not shown). Eleven best matched target genes, whose energy were lower than −26.84 kcal/mol (supplementary Table S3) were subjected to RT-PCR verification. The results showed the expression levels of these genes were not significantly changed in silkworm fed on synthetic miR166b (Fig. 2, supplementary Fig. S2).

Bottom Line: The results of TA clone, Sanger sequencing and droplet digital PCR demonstrated that several mulberry-derived miRNAs could enter to silkworm hemolymph and multiple tested tissues.Mulberry miRNAs are convincingly transferred to the silkworm orally and no physiological process associated with the miRNAs was demonstrable.The results provided a new aspect of cross-kingdom miRNA transfer.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing 400715, P. R. China.

ABSTRACT
Since a plant miRNA (miR168) cross-regulating a mammalian transcript was reported, miRNA-mediated cross-kingdom communication has become one of the most compelling but controversial topics. In the present study, we used silkworm and mulberry, which is a model for studies on the interactions between the insect and its host plant, to address whether miRNA-mediated cross-kingdom communication is a common phenomenon. The results of TA clone, Sanger sequencing and droplet digital PCR demonstrated that several mulberry-derived miRNAs could enter to silkworm hemolymph and multiple tested tissues. Synthetic miR166b was also detected in hemolymph and fat body. However, the ingestion of synthetic miR166b did not play roles in silkworm physiological progress, which was revealed by RNA-seq analyses, RT-PCR, and phenotypic investigations. Mulberry miRNAs are convincingly transferred to the silkworm orally and no physiological process associated with the miRNAs was demonstrable. The results provided a new aspect of cross-kingdom miRNA transfer.

No MeSH data available.