The Cucumber mosaic virus (CMV) Y-satellite RNA (Y-Sat) has a small non-protein-coding RNA genome that induces yellowing symptoms in infected Nicotiana tabacum (tobacco). How this RNA pathogen induces such symptoms has been a longstanding question. We show that the yellowing symptoms are a result of small interfering RNA (siRNA)-directed RNA silencing of the chlorophyll biosynthetic gene, CHLI. The CHLI mRNA contains a 22-nucleotide (nt) complementary sequence to the Y-Sat genome, and in Y-Sat-infected plants, CHLI expression is dramatically down-regulated. Small RNA sequencing and 5′ RACE analyses confirmed that this 22-nt sequence was targeted for mRNA cleavage by Y-Sat-derived siRNAs. Transformation of tobacco with a RNA interference (RNAi) vector targeting CHLI induced Y-Sat-like symptoms. In addition, the symptoms of Y-Sat infection can be completely prevented by transforming tobacco with a silencing-resistant variant of the CHLI gene. These results suggest that siRNA-directed silencing of CHLI is solely responsible for the Y-Sat-induced symptoms. Furthermore, we demonstrate that two Nicotiana species, which do not develop yellowing symptoms upon Y-Sat infection, contain a single nucleotide polymorphism within the siRNA-targeted CHLI sequence. This suggests that the previously observed species specificity of Y-Sat-induced symptoms is due to natural sequence variation in the CHLI gene, preventing CHLI silencing in species with a mismatch to the Y-Sat siRNA. Taken together, these findings provide the first demonstration of small RNA-mediated viral disease symptom production and offer an explanation of the species specificity of the viral disease.
Vyšlo v časopise: Viral Small Interfering RNAs Target Host Genes to Mediate Disease Symptoms in Plants. PLoS Pathog 7(5): e32767. doi:10.1371/journal.ppat.1002022 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002022
1. HuCCHsuYHLinNS
2009
Satellite RNAs and satellite viruses of plants.
Viruses
1
1325
1350
2. Garcia-ArenalFPalukaitisP
1999
Structure and functional relationship of satellite RNAs of
cucumber mosaic virus.
Curr Top Microbiol Immunol
239
37
63
3. PalukaitisPRoossinckMJ
1996
Spontaneous change of a benign satellite RNA of cucumber mosaic
virus to a pathogenic variant.
Nat Biotechnol
14
1264
1268
4. DevicMJaegleMBaulcombeD
1990
Cucumber mosaic virus satellite RNA (strain Y): analysis of
sequences which affect systemic necrosis on tomato.
J Gen Virol
71
1443
1449
5. SleatDEPalukaitisP
1992
A single nucleotide change within a plant virus satellite RNA
alters the host specificity of disease induction.
Plant J
2
43
49
6. ZhangLKimCHPalukaitisP
1994
The chlorosis-induction domain of the satellite RNA of cucumber
mosaic virus: identifying sequences that affect accumulation and the degree
of chlorosis.
Mol Plant Microbe Interact
7
208
213
7. WangMBWesleySVFinneganEJSmithNAWaterhousePM
2001
Replicating satellite RNA induces sequence-specific DNA
methylation and truncated transcripts in plants.
RNA
16
28
8. GhildiyalMZamorePD
Small silencing RNAs: an expanding universe.
Nat Rev Genet
10
94
108
9. WirthSCrespiM
Non-protein coding RNAs, a diverse class of gene regulators, and
their action in plants.
RNA Biol
6
161
164
10. WangMBBianXYWuLMLiuLXSmithNA
2004
On the role of RNA silencing in the pathogenicity and evolution
of viroids and viral satellites.
Proc Natl Acad Sci U S A
101
3275
3280
11. MasutaCTakanamiY
1989
Determination of sequence and structural requirements for
pathogenicity of a cucumber mosaic virus satellite RNA
(Y-satRNA).
Plant Cell
1165
1173
12. DevicMJaegleMBaulcombeD
Symptom production on tobacco and tomato is determined by two
distinct domains of the satellite RNA of cucumber mosaic virus (strain
Y).
70
2765
2774
13. JaegleMDevicMLongstaffMBaulcombeD
Cucumber mosaic virus satellite RNA (Y strain): analysis of
sequences which affect yellow mosaic symptoms on tobacco.
1905
1912
14. KuwataSMasutaCTakanamiY
1991
Reciprocal phenotype alterations between two satellite RNAs of
72
2385
2389
15. MasutaCSuzukiMKuwataSTakanamiYKoiwaiA
1993
Yellow mosaic induced by Y satellite RNA of cucumber mosaic virus
is regulated by a single incompletely dominant gene in wild
Nicotiana species.
Genetics
83
411
413
16. MasutaCSuzukiMMatsuzakiTHondaIKuwataS
Bright yellow chlorosis by cucumber mosaic virus Y satellite RNA
is specifically induced without severe chloroplast damage.
Physiol Mol Plant Pathol
42
267
278
17. HaleyBZamorePD
Kinetic analysis of the RNAi enzyme complex.
Nat Struct Mol Biol
11
599
606
18. YamaguchiKNishimuraM
2000
Reduction to below threshold levels of glycolate oxidase
activities in transgenic tobacco enhances photoinhibition during
irradiation.
Plant Cell Physiol
41
1397
1406
19. TsagrisEMMartínez de AlbaAEGozmanovaMKalantidisK
2008
Viroids.
Cell Microbiol
2168
2179
20. SchmitzARiesnerD
1998
Correlation between bending of the VM region and pathogenicity of
different Potato Spindle Tuber Viroid strains.
4
1295
1303
21. DingSWVoinnetO
2007
Antiviral immunity directed by small RNAs.
Cell
130
426
22. SleatDEPalukaitisP
Induction of tobacco chlorosis by certain cucumber mosaic virus
satellite RNAs is specific to subgroup II helper strains.
Virol
176
292
295
23. YeJQuJZhangJFGengYFFangRX
A critical domain of the Cucumber mosaic virus 2b protein for RNA
silencing suppressor activity.
FEBS Lett
583
24. ItayaAZhongXBundschuhRQiYWangY
A structured viroid RNA serves as a substrate for dicer-like
cleavage to produce biologically active small RNAs but is resistant to
RNA-induced silencing complex-mediated degradation.
J Virol
81
2980
2984
25. GottweinECullenBR
Viral and cellular microRNAs as determinants of viral
pathogenesis and immunity.
Cell Host Microbe
3
375
387
26. MoissiardGVoinnetO
2006
RNA silencing of host transcripts by cauliflower mosaic virus
requires coordinated action of the four Arabidopsis Dicer-like
proteins.
103
19593
19598
27. GreyFHookLNelsonJ
The functions of herpesvirus-encoded microRNAs.
Med Microbiol Immunol
197
261
28. QiXBaoFSXieZ
Small RNA deep sequencing reveals role for Arabidopsis thaliana
RNA-dependent RNA polymerases in viral siRNA biogenesis.
PLoS One
e4971
29. GreyFTirabassiRMeyersHWuGMcWeeneyS
2010
A viral microRNA down-regulates multiple cell cycle genes through
mRNA 5′ UTRs.
Plos Pathog
e1000967
30. LinKYChengCPChangBCWangWCHuangYW
Global analyses of small interfering RNAs derived from Bamboo
mosaic virus and its associated satellite RNAs in different
plants.
5
e11928
31. SteinhauerDAHollandJJ
1987
Rapid evolution of RNA viruses.
Ann Rev Microbiol
409
433
32. SmithNAEamensALWangMB
The presence of high-molecular-weight viral RNAs interferes with
the detection of viral small RNAs.
1062
1067
33. GleaveAP
A versatile binary vector system with a T-DNA organisational
structure conducive to efficient integration of cloned DNA into the plant
genome.
Plant Mol Biol
20
1203
1207
34. HelliwellCAWaterhousePM
2005
Constructs and methods for hairpin RNA-mediated gene silencing in
Methods Enzymol
392
24
35
35. SmithNASinghSPWangMBStoutjesdijkPAGreenAG
Total silencing by intron-spliced hairpin RNAs.
Nature
407
319
36. EllisJGLlewellynDJDennisESPeacockWJ
Maize Adh1 promoter sequences control anaerobic regulation:
Addition of upstream promoter elements from constitutive genes is necessary
for expression in tobacco.
EMBO J
Zvýšte si kvalifikáciu online z pohodlia domova
Nemáte účet? Registrujte sa
Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.
