Abstract
A tobacco calmodulin-related protein, rgs-CaM, interacts with viral suppressors of RNA silencing and modulates host RNA silencing. Plants overexpressing the rgs-CaM gene were crossed with plants exhibiting sense transgene-induced RNA silencing (S-PTGS) or inverted repeat-induced RNA silencing (IR-PTGS). S44 plants harboring a sense transgene encoding a tobacco microsomal ω-3 fatty acide desaturase (NtFAD3) exhibited the S-PTGS phenotype. The frequency of the S-PTGS phenotype incidence was nearly 100 % in the hemizygous S44 plants, but was reduced to 30 % in crossbred plants with an rgs-CaM-overexpressing transgenic line. The remaining 70 % of crossbred plants successfully overexpressed the NtFAD3 transgene, and the amount of NtFAD3 small interfering RNAs (siRNAs) was largely decreased. In contrast, overexpression of rgs-CaM did not suppress siRNA production in the IR-PTGS that targeted the NtFAD3 gene. These results indicated that rgs-CaM suppresses RNA silencing at a step upstream of siRNA production and does not interfere with the later steps of RNA silencing, including siRNA-mediated RNA degradation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Endogene:
-
Endogenous gene
- HC-Pro:
-
Helper component-proteinase
- IR-PTGS:
-
Inverted repeat-induced post-transcriptional gene silencing
- PVX:
-
Potato virus X
- siRNA:
-
Small interfering RNA
- S-PTGS:
-
Sense transgene-induced post-transcriptional gene silencing
References
Akbergenov R, Si-Ammour A, Blevins T, Amin I, Kutter C, Vanderschuren H, Zhang P, Gruissem W, Meins F Jr, Hohn T, Poogin MM (2006) Molecular characterization of geminivirus-derived small RNAs in different plant species. Nucleic Acid Res 34:462–471. doi:10.1093/nar/gki447
Anandalakshmi R, Pruss GJ, Ge X, Marathe R, Mallory AC, Smith TH, Vance VB (1998) A viral suppressor of gene silencing in plants. Proc Natl Acad Sci USA 95:13079–13084. doi:10.1073/pnas.95.22.13079
Anandalakshmi R, Marathe R, Ge X, Herr JM Jr, Mau C, Mallory A, Pruss G, Bowman L, Vance VB (2000) A calmodulin-related protein that suppresses posttranscriptional gene silencing in plants. Science 290:142–144. doi:10.1126/science.290.5489.142
Baumberger N, Baulcombe DC (2005) Arabidopsis ARGONAUTE1 is an RNA slicer that selectively recruits microRNAs and short interfering RNAs. Proc Natl Acad Sci USA 102:11928–11933. doi:10.1073/pnas.0505461102
Burgyán J, Havelda Z (2011) Viral suppressors of RNA silencing. Trends Plant Sci 16:265–272. doi:10.1016/j.tplants.2011.02.010
Dalmay T, Hamilton A, Rudd S, Angell S, Baulcombe DC (2000) An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus. Cell 101:543–553. doi:10.1016/S0092-8674(00)80864-8
Elmayan T, Balzergue S, Béon F, Bourdon V, Daubremet J, Guénet Y, Mourrain P, Palauqui JC, Vernhettes S, Vialle T, Worstrikoff K, Vaucheret H (1998) Arabidopsis mutant impaired in cosuppression. Plant Cell 10:1747–1757. doi:10.1105/tpc.10.10.1747
Fukunaga R, Doudna JA (2009) dsRNA with 5′ overhangs contributes to endogenous and antiviral RNA silencing pathways in plants. EMBO J 28:545–555. doi:10.1038/emboj.2009.2
Gazzani S, Lawrenson T, Woodward C, Headon D, Sablowski R (2004) A link between mRNA turnover and RNA interference in Arabidopsis. Science 306:1046–1048. doi:10.1126/science.1101092
Goto K, Kanazawa A, Kusaba M, Masuta C (2003) A simple and rapid method to detect plant siRNA using nonradioactive probes. Plant Mol Biol Rep 21:51–58. doi:10.1007/BF02773396
Gy I, Gasciolli V, Lauressergues D, Morel J, Gombert J, Proux F, Proux C, Vaucheret H, Mallory AC (2007) Arabidopsis FIERY1, XRN2, and XRN3 are endogenous RNA silencing suppressors. Plant Cell 19:3451–3461. doi:10.1105/tpc.107.055319
Hamada T, Kodama H, Takeshita K, Utsumi H, Iba K (1998) Characterization of transgenic tobacco with an increased α-linolenic acid level. Plant Physiol 118:591–598. doi:10.1104/pp.118.2.591
Hirai S, Oka S, Adachi E, Kodama H (2007) The effects of spacer sequences on silencing efficiency of plant RNAi vectors. Plant Cell Rep 26:651–659. doi:10.1007/s00299-006-0277-4
Hirai S, Takahashi K, Abiko T, Kodama H (2010) Loss of sense-transgene induced gene silencing by sequential introduction of the same transgene sequences in tobacco. FEBS J 277:1695–1703. doi:10.1111/j.1742-4658.2010.07591.x
Kodama H, Ito M, Ohnishi N, Suzuka I, Komamine A (1991) Molecular cloning of the gene for plant proliferating-cell nuclear antigen and expression of this gene during the cell cycle in synchronized cultures of Catharanthus roseus cells. Eur J Biochem 197:495–503. doi:10.1111/j.1432-1033.1991.tb15937.x
Kodama H, Hamada T, Horiguchi T, Nishimura M, Iba K (1994) Genetic enhancement of cold tolerance by expression of a gene for chloroplast ω-3 fatty acid desaturase in transgenic tobacco. Plant Physiol 105:601–605. doi:10.1104/pp.105.2.601
Mlotshwa S, Pruss GJ, Peragine A, Endres MW, Li J, Chen X, Poethig RS, Bowman LH, Vance V (2008) DICER-LIKE2 plays a primary role in transitive silencing of transgenes in Arabidopsis. PLoS One 3:e1755. doi:10.1371/journal.pone.0001755
Morel J, Godon C, Mourrain P, Béclin C, Boutet S, Feuerbach F, Proux F, Vaucheret H (2002) Fertile hypomorphic ARGONAUTE (ago1) mutants impaired in post-transcriptional gene silencing and virus resistance. Plant Cell 14:629–639. doi:10.1105/tpc.010358
Mourrain P, Béclin C, Elmayan T, Feuerbach F, Godon C, Morel J, Jouette D, Lacombe A, Nikic S, Picault N, Rémoué K, Sanial M, Vo T, Vaucheret H (2000) Arabidopsis SGS2 and SGS3 genes are required for posttranscriptional gene silencing and natural virus resistance. Cell 101:533–542. doi:10.1016/S0092-8674(00)80863-6
Nakahara KS, Masuta C, Yamada S, Shimura H, Kashihara Y, Wada TS, Meguro A, Goto K, Tadamura K, Sueda K, Sekiguchi T, Shao J, Itchoda N, Matsumura T, Igarashi M, Ito K, Carthew RW, Uyeda I (2012) Tobacco calmodulin-like protein provides secondary defense by binding to and directing degradation of virus RNA silencing suppressors. Proc Natl Acad Sci USA 109:10113–10118. doi:10.1073/pnas.1201628109
Napoli C, Lemieux C, Jorgensen R (1990) Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2:279–289. doi:10.1105/tpc.2.4.279
Nishimura K, Hirai S, Kodama H (2009) Effects of siRNAs targeting the spacer sequence of plant RNAi vectors on the specificity and efficiency of RNAi. J Biosci Bioeng 108:435–437. doi:10.1016/j.jbiosc.2009.05.013
Oka S, Midorikawa K, Kodama H (2010) Cosuppression and RNAi induced by Arabidopsis ortholog gene sequences in tobacco. Plant Biotechnol Rep 4:185–192. doi:10.1007/s11816-010-0135-2
Parent JS, Martinez de Alba AE, Vaucheret H (2012) The origin and effect of small RNA signaling in plants. Front Plant Sci 3:179. doi:10.3389/fpls.2012.00179
Pruss G, Ge X, Shi XM, Carrington JC, Vance VB (1997) Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell 9:859–868. doi:10.1105/tpc.9.6.859
Tadamura K, Nakahara KS, Masuta C, Uyeda I (2012) Wound-induced rgs-CaM gets ready for counterresponse to an early stage of viral infection. Plant Signal Behav 7:1548–1551. doi:10.4161/psb.22369
Tomita R, Hamada T, Horiguchi G, Iba K, Kodama H (2004) Transgene overexpression with cognate small interfering RNA in tobacco. FEBS Lett 573:117–120. doi:10.1016/j.febslet.2004.07.063
van der Krol AR, Mur LA, Beld M, Mol JNM, Stuitje AR (1990) Flavonoid genes in petunia: addition of a limited number of gene copies may lead to a suppression of a gene expression. Plant Cell 2:291–299. doi:10.1105/tpc.2.4.291
Wang MB, Masuta C, Smith NA, Shimura H (2012) RNA silencing and plant viral diseases. Mol Plant Microbe Interact 25:1275–1285. doi:10.1094/MPMI-04-12-0093-CR
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Nakamura, H., Shin, MR., Fukagawa, T. et al. A tobacco calmodulin-related protein suppresses sense transgene-induced RNA silencing but not inverted repeat-induced RNA silencing. Plant Cell Tiss Organ Cult 116, 47–53 (2014). https://doi.org/10.1007/s11240-013-0381-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-013-0381-4