A Viral Satellite RNA Induces Yellow Symptoms on Tobacco by Targeting
a Gene Involved in Chlorophyll Biosynthesis using the RNA Silencing
Machinery
Symptoms on virus-infected plants are often very specific to the given virus. The
molecular mechanisms involved in viral symptom induction have been extensively
studied, but are still poorly understood. Cucumber mosaic virus
(CMV) Y satellite RNA (Y-sat) is a non-coding subviral RNA and modifies the
typical symptom induced by CMV in specific hosts; Y-sat causes a bright yellow
mosaic on its natural host Nicotiana tabacum. The Y-sat-induced
yellow mosaic failed to develop in the infected Arabidopsis and
tomato plants suggesting a very specific interaction between Y-sat and its host.
In this study, we revealed that Y-sat produces specific short interfering RNAs
(siRNAs), which interfere with a host gene, thus inducing the specific symptom.
We found that the mRNA of tobacco magnesium protoporphyrin chelatase subunit I
(ChlI, the key gene involved in chlorophyll synthesis) had
a 22-nt sequence that was complementary to the Y-sat sequence, including four
G-U pairs, and that the Y-sat-derived siRNAs in the virus-infected plant
downregulate the mRNA of ChlI by targeting the complementary
sequence. ChlI mRNA was also downregulated in the transgenic
lines that express Y-sat inverted repeats. Strikingly, modifying the Y-sat
sequence in order to restore the 22-nt complementarity to
Arabidopsis and tomato ChlI mRNA resulted
in yellowing symptoms in Y-sat-infected Arabidopsis and tomato,
respectively. In 5′-RACE experiments, the ChlI transcript
was cleaved at the expected middle position of the 22-nt complementary sequence.
In GFP sensor experiments using agroinfiltration, we further demonstrated that
Y-sat specifically targeted the sensor mRNA containing the 22-nt complementary
sequence of ChlI. Our findings provide direct evidence that the
identified siRNAs derived from viral satellite RNA directly modulate the viral
disease symptom by RNA silencing-based regulation of a host gene.
Vyšlo v časopise:
A Viral Satellite RNA Induces Yellow Symptoms on Tobacco by Targeting
a Gene Involved in Chlorophyll Biosynthesis using the RNA Silencing
Machinery. PLoS Pathog 7(5): e32767. doi:10.1371/journal.ppat.1002021
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002021
Souhrn
Symptoms on virus-infected plants are often very specific to the given virus. The
molecular mechanisms involved in viral symptom induction have been extensively
studied, but are still poorly understood. Cucumber mosaic virus
(CMV) Y satellite RNA (Y-sat) is a non-coding subviral RNA and modifies the
typical symptom induced by CMV in specific hosts; Y-sat causes a bright yellow
mosaic on its natural host Nicotiana tabacum. The Y-sat-induced
yellow mosaic failed to develop in the infected Arabidopsis and
tomato plants suggesting a very specific interaction between Y-sat and its host.
In this study, we revealed that Y-sat produces specific short interfering RNAs
(siRNAs), which interfere with a host gene, thus inducing the specific symptom.
We found that the mRNA of tobacco magnesium protoporphyrin chelatase subunit I
(ChlI, the key gene involved in chlorophyll synthesis) had
a 22-nt sequence that was complementary to the Y-sat sequence, including four
G-U pairs, and that the Y-sat-derived siRNAs in the virus-infected plant
downregulate the mRNA of ChlI by targeting the complementary
sequence. ChlI mRNA was also downregulated in the transgenic
lines that express Y-sat inverted repeats. Strikingly, modifying the Y-sat
sequence in order to restore the 22-nt complementarity to
Arabidopsis and tomato ChlI mRNA resulted
in yellowing symptoms in Y-sat-infected Arabidopsis and tomato,
respectively. In 5′-RACE experiments, the ChlI transcript
was cleaved at the expected middle position of the 22-nt complementary sequence.
In GFP sensor experiments using agroinfiltration, we further demonstrated that
Y-sat specifically targeted the sensor mRNA containing the 22-nt complementary
sequence of ChlI. Our findings provide direct evidence that the
identified siRNAs derived from viral satellite RNA directly modulate the viral
disease symptom by RNA silencing-based regulation of a host gene.
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