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Roles and Programming of Arabidopsis ARGONAUTE Proteins during Infection


RNA silencing is a primary, adaptive defense system against viruses in plants. Viruses have evolved counter-defensive mechanisms that inhibit RNA silencing through the activity of silencing suppressor proteins. Understanding how antiviral silencing is controlled, and how suppressor proteins function, is essential for understanding how plants normally resist viruses, why some viruses are highly virulent in different hosts, and how sustainable antiviral resistance strategies can be deployed in agricultural settings. We used a mutant version of Turnip mosaic virus lacking a functional silencing suppressor (HC-Pro) to understand the genetic requirements for resistance in the model plant Arabidopsis thaliana. We focused on ARGONAUTE proteins, which have long been hypothesized to bind short interfering RNAs (siRNAs) derived from virus genomes for use as sequence-specific guides to recognize and target viral RNA for degradation or repression. We demonstrated specialized antiviral roles for specific ARGONAUTES and showed that several can bind viral siRNAs from across the entire viral genome. However, ARGONAUTE proteins are only loaded with virus-derived siRNAs in the absence of HC-Pro, which we showed binds siRNAs from the viral genome. This indicates that several AGO proteins, which collectively are necessary for full anti-TuMV defense, need to properly load virus-derived siRNAs to execute their antiviral roles.


Vyšlo v časopise: Roles and Programming of Arabidopsis ARGONAUTE Proteins during Infection. PLoS Pathog 11(3): e32767. doi:10.1371/journal.ppat.1004755
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004755

Souhrn

RNA silencing is a primary, adaptive defense system against viruses in plants. Viruses have evolved counter-defensive mechanisms that inhibit RNA silencing through the activity of silencing suppressor proteins. Understanding how antiviral silencing is controlled, and how suppressor proteins function, is essential for understanding how plants normally resist viruses, why some viruses are highly virulent in different hosts, and how sustainable antiviral resistance strategies can be deployed in agricultural settings. We used a mutant version of Turnip mosaic virus lacking a functional silencing suppressor (HC-Pro) to understand the genetic requirements for resistance in the model plant Arabidopsis thaliana. We focused on ARGONAUTE proteins, which have long been hypothesized to bind short interfering RNAs (siRNAs) derived from virus genomes for use as sequence-specific guides to recognize and target viral RNA for degradation or repression. We demonstrated specialized antiviral roles for specific ARGONAUTES and showed that several can bind viral siRNAs from across the entire viral genome. However, ARGONAUTE proteins are only loaded with virus-derived siRNAs in the absence of HC-Pro, which we showed binds siRNAs from the viral genome. This indicates that several AGO proteins, which collectively are necessary for full anti-TuMV defense, need to properly load virus-derived siRNAs to execute their antiviral roles.


Zdroje

1. Ding SW, Voinnet O. Antiviral immunity directed by small RNAs. Cell. 2007;130(3):413–26. doi: 10.1016/j.cell.2007.07.039 17693253

2. Pumplin N, Voinnet O. RNA silencing suppression by plant pathogens: defence, counter- defence and counter-counter-defence. Nat Rev Microbiol. 2013;11(11):745–60. doi: 10.1038/nrmicro3120. 24129510

3. Deleris A, Gallego-Bartolome J, Bao J, Kasschau KD, Carrington JC, Voinnet O.Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense. Science. 2006;313(5783):68–71. doi: 10.1126/science.1128214. 16741077

4. Diaz-Pendon JA, Li F, Li WX, Ding SW. Suppression of antiviral silencing by cucumber mosaic virus 2b protein in Arabidopsis is associated with drastically reduced accumulation of three classes of viral small interfering RNAs. Plant Cell. 2007;19(6):2053–63. doi: 10.1105/tpc.106.047449 17586651

5. Curtin SJ, Watson JM, Smith NA, Eamens AL, Blanchard CL, Waterhouse PM. The roles of plant dsRNA-binding proteins in RNAi-like pathways. FEBS Lett. 2008;582(18):2753–60. doi: 10.1016/j.febslet.2008.07.004 18625233

6. Qu F, Ye X, Morris TJ. Arabidopsis DRB4, AGO1, AGO7, and RDR6 participate in a DCL4-initiated antiviral RNA silencing pathway negatively regulated by DCL1. Proc Natl Acad Sci U S A. 2008;105(38):14732–7. doi: 10.1073/pnas.0805760105 18799732

7. Bologna NG, Voinnet O. The diversity, biogenesis, and activities of endogenous silencing small RNAs in Arabidopsis. Annu Rev Plant Biol. 2014;65:473–503. 10.1146/annurev- arplant-050213–035728 24579988

8. Schuck J, Gursinsky T, Pantaleo V, Burgyan J, Behrens SE. AGO/RISC-mediated antiviral RNA silencing in a plant in vitro system. Nucleic Acids Res. 2013;41(9):5090–103. 10.1093/nar/gkt193 23535144

9. Brodersen P, Sakvarelidze-Achard L, Bruun-Rasmussen M, Dunoyer P, Yamamoto YY, Sieburth L, et al. Widespread translational inhibition by plant miRNAs and siRNAs. Science. 2008;320(5880):1185–90. 10.1126/science.1159151 18483398

10. Ciomperlik JJ, Omarov RT, Scholthof HB. An antiviral RISC isolated from Tobacco rattle virus-infected plants. Virology. 2011;412(1):117–24. 10.1016/j.virol.2010.12.018 21272908

11. Iwakawa HO, Tomari Y. Molecular Insights into microRNA-Mediated Translational Repression in Plants. Mol Cell. 2013;52(4):591–601. Epub 2013/11/26. 10.1016/j.molcel.2013.10.033 24267452

12. Huntzinger E, Izaurralde E. Gene silencing by microRNAs: contributions of translational repression and mRNA decay. Nature reviews Genetics. 2011;12(2):99–110. 10.1038/nrg2936 21245828

13. Szittya G, Burgyan J. RNA interference-mediated intrinsic antiviral immunity in plants. Current topics in microbiology and immunology. 2013;371:153–81. 10.1007/978–3- 642–37765–5_6 23686235

14. Incarbone M, Dunoyer P. RNA silencing and its suppression: novel insights from in planta analyses. Trends Plant Sci. 2013;18(7):382–92. 10.1016/j.tplants.2013.04.001 23684690

15. Baumberger N, Tsai CH, Lie M, Havecker E, Baulcombe DC. The Polerovirus silencing suppressor P0 targets ARGONAUTE proteins for degradation. Curr Biol. 2007;17(18):1609–14. 10.1016/j.cub.2007.08.039 17869110

16. Bortolamiol D, Pazhouhandeh M, Marrocco K, Genschik P, Ziegler-Graff V. The Polerovirus F box protein P0 targets ARGONAUTE1 to suppress RNA silencing. Curr Biol. 2007;17(18):1615–21. 10.1016/j.cub.2007.07.061 17869109

17. Chiu MH, Chen IH, Baulcombe DC, Tsai CH. The silencing suppressor P25 of Potato virus X interacts with Argonaute1 and mediates its degradation through the proteasome pathway. Mol Plant Pathol. 2010;11(5):641–9. 10.1111/j.1364–3703.2010.00634.x 20696002

18. Csorba T, Lozsa R, Hutvagner G, Burgyan J. Polerovirus protein P0 prevents the assembly of small RNA-containing RISC complexes and leads to degradation of ARGONAUTE1. Plant J. 2010;62(3):463–72. 10.1111/j.1365–313X.2010.04163.x 20128884

19. Derrien B, Baumberger N, Schepetilnikov M, Viotti C, De Cillia J, Ziegler-Graff V, et al. Degradation of the antiviral component ARGONAUTE1 by the autophagy pathway. Proc Natl Acad Sci U S A. 2012;109(39):15942–6. 10.1073/pnas.1209487109 23019378

20. Zhang X, Yuan YR, Pei Y, Lin SS, Tuschl T, Patel DJ, et al. Cucumber mosaic virus- encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense. Genes Dev. 2006;20(23):3255–68. 10.1101/gad.1495506 17158744

21. Giner A, Lakatos L, Garcia-Chapa M, Lopez-Moya JJ, Burgyan J. Viral protein inhibits RISC activity by argonaute binding through conserved WG/GW motifs. PLoS Pathog. 2010;6(7):e1000996. 10.1371/journal.ppat.1000996 20657820

22. Nakahara KS, Masuta C. Interaction between viral RNA silencing suppressors and host factors in plant immunity. Curr Opin Plant Biol. 2014;20:88–95. 10.1016/j.pbi.2014.05.004 24875766

23. Garcia-Ruiz H, Takeda A, Chapman EJ, Sullivan CM, Fahlgren N, Brempelis KJ, et al. Arabidopsis RNA-dependent RNA polymerases and dicer-like proteins in antiviral defense and small interfering RNA biogenesis during Turnip Mosaic Virus infection. Plant Cell. 2010;22(2):481–96. 10.1105/tpc.109.073056 20190077

24. Vaucheret H. Plant ARGONAUTES. Trends Plant Sci. 2008;13(7):350–8. Epub 2008/05/30. S1360–1385(08)00138–6[pii] 10.1016/j.tplants.2008.04.007 18508405

25. Morel JB, Godon C, Mourrain P, Beclin C, Boutet S, Feuerbach F, et al. Fertile hypomorphic ARGONAUTE (ago1) mutants impaired in post-transcriptional gene silencing and virus resistance. Plant Cell. 2002;14(3):629–39. 11910010

26. Harvey JJ, Lewsey MG, Patel K, Westwood J, Heimstadt S, Carr JP, et al. An antiviral defense role of AGO2 in plants. PLoS One. 2011;6(1):e14639. 10.1371/journal.pone.0014639 21305057

27. Jaubert MJ, Bhattacharjee S, Mello AF, Perry KL, Moffett P. AGO2 mediates RNA silencing anti-viral defenses against Potato virus X in Arabidopsis. Plant physiology. 2011. Epub 2011/05/18. 10.1104/pp.111.178012

28. Zhang X, Singh J, Li D, Qu F. Temperature-dependent survival of Turnip crinkle virus- infected arabidopsis plants relies on an RNA silencing-based defense that requires dcl2, AGO2, and HEN1. Journal of virology. 2012;86(12):6847–54. Epub 2012/04/13. 10.1128/JVI.00497–12 22496240

29. Wang XB, Jovel J, Udomporn P, Wang Y, Wu Q, Li WX, et al. The 21-Nucleotide, but Not 22-Nucleotide, Viral Secondary Small Interfering RNAs Direct Potent Antiviral Defense by Two Cooperative Argonautes in Arabidopsis thaliana. The Plant cell. 2011;23(4):1625–38. Epub 2011/04/07. 10.1105/tpc.110.082305 21467580

30. Dzianott A, Sztuba-Solinska J, Bujarski JJ. Mutations in the antiviral RNAi defense pathway modify Brome mosaic virus RNA recombinant profiles. Molecular plant-microbe interactions: MPMI. 2012;25(1):97–106. Epub 2011/09/23. 10.1094/MPMI-05–11- 0137 21936664

31. Carbonell A, Fahlgren N, Garcia-Ruiz H, Gilbert KB, Montgomery TA, Nguyen T, et al. Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants. The Plant cell. 2012;24(9):3613–29. Epub 2012/10/02. 10.1105/tpc.112.099945 23023169

32. Takeda A, Iwasaki S, Watanabe T, Utsumi M, Watanabe Y. The mechanism selecting the guide strand from small RNA duplexes is different among argonaute proteins. Plant Cell Physiol. 2008;49(4):493–500. 10.1093/pcp/pcn043 18344228

33. Azevedo J, Garcia D, Pontier D, Ohnesorge S, Yu A, Garcia S, et al. Argonaute quenching and global changes in Dicer homeostasis caused by a pathogen-encoded GW repeat protein. Genes Dev. 2010;24(9):904–15. 10.1101/gad.1908710 20439431

34. Wei W, Ba Z, Gao M, Wu Y, Ma Y, Amiard S, et al. A role for small RNAs in DNA double- strand break repair. Cell. 2012;149(1):101–12. Epub 2012/03/27. 10.1016/j.cell.2012.03.002 22445173

35. Zhang X, Zhao H, Gao S, Wang WC, Katiyar-Agarwal S, Huang HD, et al. Arabidopsis Argonaute 2 regulates innate immunity via miRNA393 (*)-mediated silencing of a Golgi- localized SNARE gene, MEMB12. Mol Cell. 2011;42(3):356–66. 10.1016/j.molcel.2011.04.010 21549312

36. Zhu H, Hu F, Wang R, Zhou X, Sze SH, Liou LW, et al. Arabidopsis Argonaute10 specifically sequesters miR166/165 to regulate shoot apical meristem development. Cell. 2011;145(2):242–56. 10.1016/j.cell.2011.03.024 21496644

37. Mallory AC, Hinze A, Tucker MR, Bouche N, Gasciolli V, Elmayan T, et al. Redundant and specific roles of the ARGONAUTE proteins AGO1 and ZLL in development and small RNA-directed gene silencing. PLoS Genet. 2009;5(9):e1000646. 10.1371/journal.pgen.1000646 19763164

38. Kasschau KD, Cronin S, Carrington JC. Genome amplification and long-distance movement functions associated with the central domain of tobacco etch potyvirus helper component-proteinase. Virology. 1997;228(2):251–62. 10.1006/viro.1996.8368 9123832

39. Lakatos L, Csorba T, Pantaleo V, Chapman EJ, Carrington JC, Liu YP, et al. Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors. EMBO J. 2006;25(12):2768–80. 10.1038/sj.emboj.7601164 16724105

40. Mallory AC, Reinhart BJ, Bartel D, Vance VB, Bowman LH. A viral suppressor of RNA silencing differentially regulates the accumulation of short interfering RNAs and micro- RNAs in tobacco. Proc Natl Acad Sci U S A. 2002;99(23):15228–33. 10.1073/pnas.232434999 12403829

41. Chapman EJ, Prokhnevsky AI, Gopinath K, Dolja VV, Carrington JC. Viral RNA silencing suppressors inhibit the microRNA pathway at an intermediate step. Genes Dev. 2004;18(10):1179–86. 10.1101/gad.1201204 15131083

42. Kasschau KD, Xie Z, Allen E, Llave C, Chapman EJ, Krizan KA, et al. P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA unction. Dev Cell. 2003;4(2):205–17. 12586064

43. Schott G, Mari-Ordonez A, Himber C, Alioua A, Voinnet O, Dunoyer P. Differential effects of viral silencing suppressors on siRNA and miRNA loading support the existence of two distinct cellular pools of ARGONAUTE1. The EMBO journal. 2012;31(11):2553–65. Epub 2012/04/26. 10.1038/emboj.2012.92 22531783

44. Shiboleth YM, Haronsky E, Leibman D, Arazi T, Wassenegger M, Whitham SA, et al. Theconserved FRNK box in HC-Pro, a plant viral suppressor of gene silencing, is required for small RNA binding and mediates symptom development. J Virol. 2007;81(23):13135–48. 10.1128/JVI.01031–07 17898058

45. Endres MW, Gregory BD, Gao Z, Foreman AW, Mlotshwa S, Ge X, et al. Two plant viral suppressors of silencing require the ethylene-inducible host transcription factor RAV2 to block RNA silencing. PLoS Pathog. 2010;6(1):e1000729. 10.1371/journal.ppat.1000729 20084269

46. Ala-Poikela M, Goytia E, Haikonen T, Rajamaki ML, Valkonen JP. Helper component proteinase of the genus Potyvirus is an interaction partner of translation initiation factors eIF(iso)4E and eIF4E and contains a 4E binding motif. J Virol. 2011;85(13):6784–94. 10.1128/JVI.00485–11 21525344

47. Anandalakshmi R, Marathe R, Ge X, Herr JM Jr., Mau C, Mallory A, et al. A calmodulin- related protein that suppresses posttranscriptional gene silencing in plants. Science. 2000;290(5489):142–4. Epub 2000/10/06. 11021800

48. Iki T, Yoshikawa M, Nishikiori M, Jaudal MC, Matsumoto-Yokoyama E, Mitsuhara I, et al. In vitro assembly of plant RNA-induced silencing complexes facilitated by molecular chaperone HSP90. Mol Cell. 2010;39(2):282–91. 10.1016/j.molcel.2010.05.014 20605502

49. Ballut L, Drucker M, Pugniere M, Cambon F, Blanc S, Roquet F, et al. HcPro, a multifunctional protein encoded by a plant RNA virus, targets the 20S proteasome and affects its enzymic activities. J Gen Virol. 2005;86(Pt 9):2595–603. 10.1099/vir.0.81107–0 16099919

50. Soitamo AJ, Jada B, Lehto K. HC-Pro silencing suppressor significantly alters the gene expression profile in tobacco leaves and flowers. BMC Plant Biol. 2011;11:68. 10.1186/1471–2229–11–68 21507209

51. Lellis AD, Kasschau KD, Whitham SA, Carrington JC. Loss-of-susceptibility mutants of Arabidopsis thaliana reveal an essential role for eIF(iso)4E during potyvirus infection. Curr Biol. 2002;12(12):1046–51. Epub 2002/07/19. 12123581

52. Montgomery TA, Howell MD, Cuperus JT, Li D, Hansen JE, Alexander AL, et al. Specificity of ARGONAUTE7-miR390 interaction and dual functionality in TAS3 trans- acting siRNA formation. Cell. 2008;133(1):128–41. Epub 2008/03/18. 10.1016/j.cell.2008.02.033 18342362

53. Mi S, Cai T, Hu Y, Chen Y, Hodges E, Ni F, et al. Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5' terminal nucleotide. Cell. 2008;133(1):116–27. 10.1016/j.cell.2008.02.034 18342361

54. Wang H, Zhang X, Liu J, Kiba T, Woo J, Ojo T, et al. Deep sequencing of small RNAs specifically associated with Arabidopsis AGO1 and AGO4 uncovers new AGO functions. Plant J. 2011;67(2):292–304. Epub 2011/04/05. 10.1111/j.1365–313X.2011.04594.x 21457371

55. Merai Z, Kerenyi Z, Kertesz S, Magna M, Lakatos L, Silhavy D. Double-stranded RNA binding may be a general plant RNA viral strategy to suppress RNA silencing. J Virol. 2006;80(12):5747–56. 10.1128/JVI.01963–05 16731914

56. Cao M, Du P, Wang X, Yu YQ, Qiu YH, Li W, et al. Virus infection triggers widespread silencing of host genes by a distinct class of endogenous siRNAs in Arabidopsis. Proc Natl Acad Sci U S A. 2014;111(40):14613–8. 10.1073/pnas.1407131111 25201959

57. Omarov RT, Ciomperlik JJ, Scholthof HB. RNAi-associated ssRNA-specific ribonucleases in Tombusvirus P19 mutant-infected plants and evidence for a discrete siRNA-containing effector complex. Proc Natl Acad Sci U S A. 2007;104(5):1714–9. 10.1073/pnas.0608117104 17244709

58. Pantaleo V, Szittya G, Burgyan J. Molecular bases of viral RNA targeting by viral small interfering RNA-programmed RISC. J Virol. 2007;81(8):3797–806. 10.1128/JVI.02383–06 17267504

59. Schmid M, Davison TS, Henz SR, Pape UJ, Demar M, Vingron M, et al. A gene expression map of Arabidopsis thaliana development. Nat Genet. 2005;37(5):501–6. 10.1038/ng1543 15806101

60. Baumberger N, Baulcombe DC. Arabidopsis ARGONAUTE1 is an RNA Slicer that selectively recruits microRNAs and short interfering RNAs. Proc Natl Acad Sci U S A. 2005;102(33):11928–33. 10.1073/pnas.0505461102 16081530

61. Donaire L, Barajas D, Martinez-Garcia B, Martinez-Priego L, Pagan I, Llave C. Structural and genetic requirements for the biogenesis of tobacco rattle virus-derived small interfering RNAs. J Virol. 2008;82(11):5167–77. 10.1128/JVI.00272–08 18353962

62. Cao M, Ye X, Willie K, Lin J, Zhang X, Redinbaugh MG, et al. The capsid protein of Turnip crinkle virus overcomes two separate defense barriers to facilitate systemic movement of the virus in Arabidopsis. J Virol. 2010;84(15):7793–802. 10.1128/JVI.02643–09 20504923

63. Blevins T, Rajeswaran R, Shivaprasad PV, Beknazariants D, Si-Ammour A, Park HS, et al. Four plant Dicers mediate viral small RNA biogenesis and DNA virus induced silencing. Nucleic Acids Res. 2006;34(21):6233–46. 10.1093/nar/gkl886 17090584

64. Wang N, Zhang D, Wang Z, Xun H, Ma J, Wang H, et al. Mutation of the RDR1 gene caused genome-wide changes in gene expression, regional variation in small RNA clusters and localized alteration in DNA methylation in rice. BMC Plant Biol. 2014;14:177. 10.1186/1471–2229–14–177 24980094

65. Montgomery TA, Yoo SJ, Fahlgren N, Gilbert SD, Howell MD, Sullivan CM, et al. AGO1- miR173 complex initiates phased siRNA formation in plants. Proc Natl Acad Sci U S A. 2008;105(51):20055–62. Epub 2008/12/11. 0810241105[pii]10.1073/pnas.0810241105 19066226

66. Cuperus JT, Carbonell A, Fahlgren N, Garcia-Ruiz H, Burke RT, Takeda A, et al. Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis. Nature structural & molecular biology. 2010;17(8):997–1003. Epub 2010/06/22. 10.1038/nsmb.1866

67. Rajeswaran R, Aregger M, Zvereva AS, Borah BK, Gubaeva EG, Pooggin MM. Sequencing of RDR6-dependent double-stranded RNAs reveals novel features of plant siRNA biogenesis. Nucleic Acids Res. 2012;40(13):6241–54. 10.1093/nar/gks242 22434877

68. Chen HM, Chen LT, Patel K, Li YH, Baulcombe DC, Wu SH. 22-Nucleotide RNAs trigger secondary siRNA biogenesis in plants. Proc Natl Acad Sci U S A. 2010;107(34):15269–74. 10.1073/pnas.1001738107 20643946

69. Allen E, Xie Z, Gustafson AM, Carrington JC. microRNA-directed phasing during trans- acting siRNA biogenesis in plants. Cell. 2005;121(2):207–21. 10.1016/j.cell.2005.04.004 15851028

70. Bhattacharjee S, Zamora A, Azhar MT, Sacco MA, Lambert LH, Moffett P. Virus resistance induced by NB-LRR proteins involves Argonaute4-dependent translational control. The Plant journal: for cell and molecular biology. 2009;58(6):940–51. Epub 2009/02/18. 10.1111/j.1365–313X.2009.03832.x

71. Curtis MD, Grossniklaus U. A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol. 2003;133(2):462–9. 10.1104/pp.103.027979 14555774

72. Lobbes D, Rallapalli G, Schmidt DD, Martin C, Clarke J. SERRATE: a new player on the plant microRNA scene. EMBO Rep. 2006;7(10):1052–8. 10.1038/sj.embor.7400806 16977334

73. Agorio A, Vera P. ARGONAUTE4 is required for resistance to Pseudomonas syringae in Arabidopsis. Plant Cell. 2007;19(11):3778–90. 10.1105/tpc.107.054494 17993621

74. Hunter C, Sun H, Poethig RS. The Arabidopsis heterochronic gene ZIPPY is an ARGONAUTE family member. Curr Biol. 2003;13(19):1734–9. 14521841

75. Kleinboelting N, Huep G, Kloetgen A, Viehoever P, Weisshaar B. GABI-Kat SimpleSearch: new features of the Arabidopsis thaliana T-DNA mutant database. Nucleic Acids Res. 2012;40(Database issue):D1211–5. 10.1093/nar/gkr1047 22080561

76. Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, et al. Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science. 2003;301(5633):653–7. 10.1126/science.1086391 12893945

77. Clough SJ, Bent AF. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 1998;16(6):735–43. 10069079

78. Varallyay E, Valoczi A, Agyi A, Burgyan J, Havelda Z. Plant virus-mediated induction of miR168 is associated with repression of ARGONAUTE1 accumulation. EMBO J. 2010;29(20):3507–19. 10.1038/emboj.2010.215 20823831

79. Gilbert K, Faber N, Kasschau K, Chapman EJ, Carrington JC, Carbonell A. Preparation of Multiplexed Small RNA Libraries From Plants. Bioprotocol. 2014;4(21).

80. Fahlgren N, Sullivan CM, Kasschau KD, Chapman EJ, Cumbie JS, Montgomery TA, et al. Computational and analytical framework for small RNA profiling by high-throughput sequencing. RNA. 2009;15(5):992–1002. 10.1261/rna.1473809 19307293

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