Quantitative estimation of the effects of propionylshikonin on the binding of TMV RNA and tobacco mRNA to wheat germ ribosome in vitro

A method involving reverse transcription and real-time polymerase chain reaction (PCR) was developed in this study to detect the effects of the antiviral compound propionylshikonin on the binding of tobacco mosaic virus (TMV) RNA and tobacco mRNA to wheat germ ribosome in vitro. TMV RNA–wheat germ ribosome and tobacco mRNA–wheat germ ribosome binding systems were constructed, and the TMV RNA–ribosome and tobacco mRNA–ribosome complexes were isolated from the binding systems using 30% sucrose cushion. The target genes for the quantitative detection of TMV RNA and tobacco mRNA were the TMV coat protein gene and tobacco elongation factor-1α gene, respectively. The designed protocol was efficient for rapid and conclusive determination of the variations in the bound TMV RNA and tobacco mRNA from the complexes with and without propionylshikonin. The inhibition rates, ranging from 26.4% to 63.6%, were detected in the bound TMV RNA with 2–10 μg/mL propionylshikonin in the binding systems. The amount of bound tobacco mRNA did not decrease in the presence of propionylshikonin, indicating that propionylshikonin did not inhibit the binding of tobacco mRNA to wheat germ ribosome. To the best of our knowledge, this is the first study on the interactions among an anti-TMV agent, TMV RNA, and a host using real-time PCR to be reported.

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► TMVRNA or tobacco mRNA reacts with wheat germ ribosome in vitro.
► The bound RNA was detected by centrifugation/RT-PCR method.
► TMV coat protein gene or tobacco elongation factor-1α gene is as target genes for RT-PCR.
► Propionylshikonin inhibits TMV RNA binding with wheat germ ribosome.
► Propionylshikonin does not inhibit tobacco mRNA binding with wheat germ ribosome.