#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

The miR9863 Family Regulates Distinct Alleles in Barley to Attenuate NLR Receptor-Triggered Disease Resistance and Cell-Death Signaling


Plants rely on cell-surface and intracellular immune receptors to sense pathogen invasion and to mediate defense responses. However, uncontrolled activation of immune responses is harmful to plant growth and development. Small RNAs have recently been shown to fine-tune the expression of intracellular immune receptors and contribute to the regulation of defense signaling in dicot plants, while similar processes have not been well documented in monocot grain crops, such as barley and wheat. Here, we show that, in barley, some members of the miR9863 family target a subset of Mla alleles that confer race-specific disease resistance to the powdery mildew fungus. These miRNAs act on Mla transcripts by cleavage and translational repression. Production of a type of trans-acting small RNAs, designated as phasiRNAs, enhances the effects of miRNA regulation on Mla targets. We propose that Mla-mediated immune signaling is fine-tuned by the miRNAs at later stage of MLA activation to avoid overloading of immune responses in barley cells.


Vyšlo v časopise: The miR9863 Family Regulates Distinct Alleles in Barley to Attenuate NLR Receptor-Triggered Disease Resistance and Cell-Death Signaling. PLoS Genet 10(12): e32767. doi:10.1371/journal.pgen.1004755
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004755

Souhrn

Plants rely on cell-surface and intracellular immune receptors to sense pathogen invasion and to mediate defense responses. However, uncontrolled activation of immune responses is harmful to plant growth and development. Small RNAs have recently been shown to fine-tune the expression of intracellular immune receptors and contribute to the regulation of defense signaling in dicot plants, while similar processes have not been well documented in monocot grain crops, such as barley and wheat. Here, we show that, in barley, some members of the miR9863 family target a subset of Mla alleles that confer race-specific disease resistance to the powdery mildew fungus. These miRNAs act on Mla transcripts by cleavage and translational repression. Production of a type of trans-acting small RNAs, designated as phasiRNAs, enhances the effects of miRNA regulation on Mla targets. We propose that Mla-mediated immune signaling is fine-tuned by the miRNAs at later stage of MLA activation to avoid overloading of immune responses in barley cells.


Zdroje

1. JonesJD, DanglJL (2006) The plant immune system. Nature 444: 323–329.

2. ZipfelC (2009) Early molecular events in PAMP-triggered immunity. Curr Opin Plant Biol 12: 414–420.

3. DoddsPN, RathjenJP (2010) Plant immunity: towards an integrated view of plant–pathogen interactions. Nat Rev Genet 11: 539–548.

4. MaekawaT, KuferTA, Schulze-LefertP (2011) NLR functions in plant and animal immune systems: so far and yet so close. Nat Immunol 12: 817–826.

5. TsudaK, SatoM, StoddardT, GlazebrookJ, KatagiriF (2009) Network properties of robust immunity in plants. PLoS Genet 5: e1000772.

6. MeyersBC, KaushikS, NandetyRS (2005) Evolving disease resistance genes. Curr Opin Plant Biol 8: 129–134.

7. TianD, TrawMB, ChenJQ, KreitmanM, BergelsonJ (2003) Fitness costs of R-gene-mediated resistance in Arabidopsis thaliana. Nature 423: 74–77.

8. BombliesK, LempeJ, EppleP, WarthmannN, LanzC, et al. (2007) Autoimmune response as a mechanism for a dobzhansky-muller-type incompatibility syndrome in plants. PLoS Biol 5: e236.

9. Jones-RhoadesMW, BartelDP, BartelB (2006) MicroRNAs and their regulatory roles in plants. Annu Rev Plant Biol 57: 19–53.

10. MalloryA, VaucheretH (2006) Functions of microRNAs and related small RNAs in plants. Nat Genet 38: S31–S36.

11. DingSW (2010) RNA-based antiviral immunity. Nat Rev Immunol 10: 632–644.

12. Katiyar-AgarwalS, JinH (2010) Role of small RNAs in host-microbe interactions. Annu Rev Phytopathol 48: 225–246.

13. PumplinN, VoinnetO (2013) RNA silencing suppression by plant pathogens: defence, counter-defence and counter-counter-defence. Nat Rev Micro 11: 745–760.

14. NavarroL, DunoyerP, JayF, ArnoldB, DharmasiriN, et al. (2006) A plant miRNA contributes to antibacterial resistance by repressing auxin signaling. Science 312: 436–439.

15. NavarroL, JayF, NomuraK, HeSY, VoinnetO (2008) Suppression of the microRNA pathway by bacterial effector proteins. Science 321: 964–967.

16. ZhangX, ZhaoH, GaoS, WangW-C, Katiyar-AgarwalS, et al. (2011) Arabidopsis argonaute 2 regulates innate immunity via miRNA393*-mediated silencing of a golgi-localized SNARE gene, MEMB12. Mol Cell 42: 356–366.

17. XuW, MengY, WiseRP (2014) Mla- and Rom1-mediated control of microRNA398 and chloroplast copper/zinc superoxide dismutase regulates cell death in response to the barley powdery mildew fungus. New Phytol 201: 1396–1412.

18. Katiyar-AgarwalS, GaoS, Vivian-SmithA, JinH (2007) A novel class of bacteria-induced small RNAs in Arabidopsis. Genes Dev 21: 3123–3134.

19. Katiyar-AgarwalS, MorganR, DahlbeckD, BorsaniO, VillegasAJr, et al. (2006) A pathogen-inducible endogenous siRNA in plant immunity. Proc Natl Acad Sci U S A 103: 18002–18007.

20. EllendorffU, FradinEF, de JongeR, ThommaBPHJ (2009) RNA silencing is required for Arabidopsis defence against Verticillium wilt disease. J Exp Bot 60: 591–602.

21. QiaoY, LiuL, XiongQ, FloresC, WongJ, et al. (2013) Oomycete pathogens encode RNA silencing suppressors. Nat Genet 45: 330–333.

22. WeibergA, WangM, LinF-M, ZhaoH, ZhangZ, et al. (2013) Fungal small RNAs suppress plant immunity by hijacking host RNA interference pathways. Science 342: 118–123.

23. BaulcombeD (2004) RNA silencing in plants. Nature 431: 356–363.

24. RogersK, ChenX (2013) Biogenesis, turnover, and mode of action of plant microRNAs. Plant Cell 25: 2383–2399.

25. MalloryA, VaucheretH (2010) Form, function, and regulation of ARGONAUTE proteins. Plant Cell 22: 3879–3889.

26. VoinnetO (2009) Origin, biogenesis, and activity of plant microRNAs. Cell 136: 669–687.

27. TangG, ReinhartBJ, BartelDP, ZamorePD (2003) A biochemical framework for RNA silencing in plants. Genes Dev 17: 49–63.

28. BaumbergerN, BaulcombeDC (2005) Arabidopsis ARGONAUTE1 is an RNA slicer that selectively recruits microRNAs and short interfering RNAs. Proc Natl Acad Sci U S A 102: 11928–11933.

29. LlaveC, XieZ, KasschauKD, CarringtonJC (2002) Cleavage of scarecrow-like mRNA targets directed by a class of Arabidopsis miRNA. Science 297: 2053–2056.

30. AukermanM, SakaiH (2003) Regulation of flowering time and floral organ identity by a microRNA and its APETALA2-like target genes. Plant Cell 15: 2730–2741.

31. ChenX (2004) A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development. Science 303: 2022–2025.

32. BrodersenP, Sakvarelidze-AchardL, Bruun-RasmussenM, DunoyerP, YamamotoYY, et al. (2008) Widespread translational inhibition by plant miRNAs and siRNAs. Science 320: 1185–1190.

33. DugasD, BartelB (2008) Sucrose induction of Arabidopsis miR398 represses two Cu/Zn superoxide dismutases. Plant Mol Biol 67: 403–417.

34. LanetE, DelannoyE, SormaniR, FlorisM, BrodersenP, et al. (2009) Biochemical evidence for translational repression by Arabidopsis microRNAs. Plant Cell 21: 1762–1768.

35. BeauclairL, YuA, BouchéN (2010) microRNA-directed cleavage and translational repression of the copper chaperone for superoxide dismutase mRNA in Arabidopsis. Plant J 62: 454–462.

36. GandikotaM, BirkenbihlR, HohmannS, CardonG, SaedlerH, et al. (2007) The miRNA156/157 recognition element in the 3' UTR of the Arabidopsis SBP box gene SPL3 prevents early flowering by translational inhibition in seedlings. Plant J 49: 683–693.

37. LiS, LiuL, ZhuangX, YuY, LiuX, et al. (2013) MicroRNAs inhibit the translation of target mRNAs on the endoplasmic reticulum in Arabidopsis. Cell 153: 562–574.

38. ZhaiJ, JeongDH, De PaoliE, ParkS, RosenBD, et al. (2011) MicroRNAs as master regulators of the plant NB-LRR defense gene family via the production of phased, trans-acting siRNAs. Genes Dev 25: 2540–2553.

39. LiF, PignattaD, BendixC, BrunkardJO, CohnMM, et al. (2012) MicroRNA regulation of plant innate immune receptors. Proc Natl Acad Sci U S A 109: 1790–1795.

40. ShivaprasadPV, ChenHM, PatelK, BondDM, SantosBA, et al. (2012) A microRNA superfamily regulates nucleotide binding site-leucine-rich repeats and other mRNAs. Plant Cell 24: 859–874.

41. BoccaraM, SarazinA, ThiebeauldO, JayF, VoinnetO, et al. (2014) The Arabidopsis miR472-RDR6 silencing pathway modulates PAMP- and effector-triggered immunity through the post-transcriptional control of disease resistance genes. PLoS Pathog 10: e1003883.

42. KällmanT, ChenJ, GyllenstrandN, LagercrantzU (2013) A significant fraction of 21-nucleotide small RNA originates from phased degradation of resistance genes in several perennial species. Plant Physiol 162: 741–754.

43. FeiQ, XiaR, MeyersBC (2013) Phased, secondary, small interfering RNAs in posttranscriptional regulatory networks. Plant Cell 25: 2400–2415.

44. ChenHM, ChenLT, PatelK, LiYH, BaulcombeDC, et al. (2010) 22-Nucleotide RNAs trigger secondary siRNA biogenesis in plants. Proc Natl Acad Sci U S A 107: 15269–15274.

45. CuperusJT, CarbonellA, FahlgrenN, Garcia-RuizH, BurkeRT, et al. (2010) Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis. Nat Struct Mol Biol 17: 997–1003.

46. ZhaiJ, ZhaoY, SimonSA, HuangS, PetschK, et al. (2013) Plant microRNAs display differential 3′ truncation and tailing modifications that are ARGONAUTE1 dependent and conserved across species. Plant Cell 25: 2417–2428.

47. ManavellaPA, KoenigD, WeigelD (2012) Plant secondary siRNA production determined by microRNA-duplex structure. Proc Natl Acad Sci U S A 109: 2461–2466.

48. SeeholzerS, TsuchimatsuT, JordanT, BieriS, PajonkS, et al. (2010) Diversity at the Mla powdery mildew resistance locus from cultivated barley reveals sites of positive selection. Mol Plant Microbe Interact 23: 497–509.

49. ShenQH, Schulze-LefertP (2007) Rumble in the nuclear jungle: compartmentalization, trafficking, and nuclear action of plant immune receptors. EMBO J 26: 4293–4301.

50. ChangC, ZhangL, ShenQH (2013) Partitioning, repressing and derepressing: dynamic regulations in MLA immune receptor triggered defense signaling. Front Plant Sci 4: 396.

51. CaldoRA, NettletonD, WiseRP (2004) Interaction-dependent gene expression in Mla-specified response to barley powdery mildew. Plant Cell 16: 2514–2528.

52. ShenQH, SaijoY, MauchS, BiskupC, BieriS, et al. (2007) Nuclear activity of MLA immune receptors links isolate-specific and basal disease-resistance responses. Science 315: 1098–1103.

53. MengY, WiseRP (2012) HvWRKY10, HvWRKY19, and HvWRKY28 regulate Mla-triggered immunity and basal defense to barley powdery mildew. Mol Plant Microbe Interact 25: 1492–1505.

54. ChangC, YuD, JiaoJ, JingS, Schulze-LefertP, et al. (2013) Barley MLA immune receptors directly interfere with antagonistically acting transcription factors to initiate disease resistance signaling. Plant Cell 25: 1158–1173.

55. MoscouMJ, LauterN, CaldoRA, NettletonD, WiseRP (2011) Quantitative and temporal definition of the Mla transcriptional regulon during barley–powdery mildew interactions. Mol Plant Microbe Interact 24: 694–705.

56. BaiS, LiuJ, ChangC, ZhangL, MaekawaT, et al. (2012) Structure-function analysis of barley NLR immune receptor MLA10 reveals its cell compartment specific activity in cell death and disease resistance. PLoS Pathog 8: e1002752.

57. MaekawaT, KracherB, VernaldiS, Ver Loren van ThemaatE, Schulze-LefertP (2012) Conservation of NLR-triggered immunity across plant lineages. Proc Natl Acad Sci U S A 109: 20119–20123.

58. MaekawaT, ChengW, SpiridonLN, TollerA, LukasikE, et al. (2011) Coiled-coil domain-dependent homodimerization of intracellular barley immune receptors defines a minimal functional module for triggering cell death. Cell Host Microbe 9: 187–199.

59. HaltermanDA, WiseRP (2004) A single-amino acid substitution in the sixth leucine-rich repeat of barley MLA6 and MLA13 alleviates dependence on RAR1 for disease resistance signaling. Plant J 38: 215–226.

60. ShirasuK (2009) The HSP90-SGT1 chaperone complex for NLR immune sensors. Annu Rev Plant Biol 60: 139–164.

61. BieriS, MauchS, ShenQH, PeartJ, DevotoA, et al. (2004) RAR1 positively controls steady state levels of barley MLA resistance proteins and enables sufficient MLA6 accumulation for effective resistance. Plant Cell 16: 3480–3495.

62. KølsterP, MunkL, StølenO, LøhdeJ (1986) Near-Isogenic Barley Lines with Genes for Resistance to Powdery Mildew. Crop Sci 26: 903–907.

63. Mahalingam G, Meyers B (2010) Computational methods for comparative analysis of plant small RNAs. In: Meyers BC, Green PJ, editors. Plant MicroRNAs: Humana Press. pp. 163–181.

64. SchreiberA, ShiB-J, HuangC-Y, LangridgeP, BaumannU (2011) Discovery of barley miRNAs through deep sequencing of short reads. BMC Genomics 12: 129–149.

65. HackenbergM, ShiBJ, GustafsonP, LangridgeP (2012) A transgenic transcription factor (TaDREB3) in barley affects the expression of microRNAs and other small non-coding RNAs. PLoS One 7: e42030.

66. WeiB, CaiT, ZhangR, LiA, HuoN, et al. (2009) Novel microRNAs uncovered by deep sequencing of small RNA transcriptomes in bread wheat (Triticum aestivum L.) and Brachypodium distachyon (L.) Beauv. Funct Int Genomics 9: 499–511.

67. KozomaraA, Griffiths-JonesS (2011) miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res 39: D152–D157.

68. MeyersBC, AxtellMJ, BartelB, BartelDP, BaulcombeD, et al. (2008) Criteria for annotation of plant microRNAs. Plant Cell 20: 3186–3190.

69. PatersonAH, BowersJE, BruggmannR, DubchakI, GrimwoodJ, et al. (2009) The Sorghum bicolor genome and the diversification of grasses. Nature 457: 551–556.

70. BertoliniE, VerelstW, HornerDS, GianfranceschiL, PiccoloV, et al. (2013) Addressing the Role of microRNAs in Reprogramming Leaf Growth during Drought Stress in Brachypodium distachyon. Molecular Plant 6: 423–443.

71. JeongD-H, SchmidtS, RymarquisL, ParkS, GanssmannM, et al. (2013) Parallel analysis of RNA ends enhances global investigation of microRNAs and target RNAs of Brachypodium distachyon. Genome Biology 14: R145.

72. JeongDH, ThatcherSR, BrownRS, ZhaiJ, ParkS, et al. (2013) Comprehensive investigation of microRNAs enhanced by analysis of sequence variants, expression patterns, ARGONAUTE loading, and target cleavage. Plant Physiol 162: 1225–1245.

73. EhrenreichIM, PuruggananMD (2008) Sequence variation of MicroRNAs and their binding sites in Arabidopsis. Plant Physiol 146: 1974–1982.

74. SchwabR, OssowskiS, RiesterM, WarthmannN, WeigelD (2006) Highly specific gene silencing by artificial microRNAs in Arabidopsis. Plant Cell 18: 1121–1133.

75. HaltermanD, ZhouF, WeiF, WiseRP, Schulze-LefertP (2001) The MLA6 coiled-coil, NBS-LRR protein confers AvrMla6-dependent resistance specificity to Blumeria graminis f. sp. hordei in barley and wheat. Plant J 25: 335–348.

76. HaltermanDA, WeiF, WiseRP (2003) Powdery mildew-induced Mla mRNAs are alternatively spliced and contain multiple upstream open reading frames. Plant Physiol 131: 558–567.

77. ShenQH, ZhouF, BieriS, HaizelT, ShirasuK, et al. (2003) Recognition specificity and RAR1/SGT1 dependence in barley Mla disease resistance genes to the powdery mildew fungus. Plant Cell 15: 732–744.

78. ZhouF, KurthJ, WeiF, ElliottC, ValeG, et al. (2001) Cell-autonomous expression of barley Mla1 confers race-specific resistance to the powdery mildew fungus via a Rar1-independent signaling pathway. Plant Cell 13: 337–350.

79. YuanC, LiC, YanL, JacksonAO, LiuZ, et al. (2011) A high throughput barley stripe mosaic virus vector for virus induced gene silencing in monocots and dicots. PLoS One 6: e26468.

80. YanJ, GuY, JiaX, KangW, PanS, et al. (2012) Effective small RNA destruction by the expression of a short tandem target mimic in Arabidopsis. Plant Cell 24: 415–427.

81. MiS, CaiT, HuY, ChenY, HodgesE, et al. (2008) Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5' terminal nucleotide. Cell 133: 116–127.

82. YeR, WangW, IkiT, LiuC, WuY, et al. (2012) Cytoplasmic assembly and selective nuclear import of arabidopsis ARGONAUTE4/siRNA complexes. Mol Cell 46: 859–870.

83. JonesL, KeiningT, EamensA, VaistijFE (2006) Virus-induced gene silencing of argonaute genes in Nicotiana benthamiana demonstrates that extensive systemic silencing requires Argonaute1-like and Argonaute4-like genes. Plant Physiol 141: 598–606.

84. LiJF, ChungHS, NiuY, BushJ, McCormackM, et al. (2013) Comprehensive protein-based artificial microRNA screens for effective gene silencing in plants. Plant Cell 25: 1507–1522.

85. PanstrugaR (2004) A golden shot: how ballistic single cell transformation boosts the molecular analysis of cereal–mildew interactions. Mol Plant Pathol 5: 141–148.

86. CaldoRA, NettletonD, PengJ, WiseRP (2006) Stage-specific suppression of basal defense discriminates barley plants containing fast- and delayed-acting Mla powdery mildew resistance alleles. Mol Plant Microbe Interact 19: 939–947.

87. MaC, LuY, BaiS, ZhangW, DuanX, et al. (2014) Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted NBS–LRR Protein Class Gene in Apple (Golden Delicious). Mol Plant 7: 218–230.

88. LvS, NieX, WangL, DuX, BiradarSS, et al. (2012) Identification and characterization of microRNAs from barley (Hordeum vulgare L.) by high-throughput sequencing. Int J Mol Sci 13: 2973–2984.

89. PeriyannanS, MooreJ, AyliffeM, BansalU, WangX, et al. (2013) The gene Sr33, an ortholog of barley Mla genes, encodes resistance to wheat stem rust race Ug99. Science 341: 786–788.

90. JordanT, SeeholzerS, SchwizerS, TollerA, SomssichIE, et al. (2011) The wheat Mla homologue TmMla1 exhibits an evolutionarily conserved function against powdery mildew in both wheat and barley. Plant J 65: 610–621.

91. GuoX, GuiY, WangY, ZhuQ-H, HelliwellC, et al. (2008) Selection and mutation on microRNA target sequences during rice evolution. BMC Genomics 9: 454.

92. ChenK, RajewskyN (2006) Natural selection on human microRNA binding sites inferred from SNP data. Nat Genet 38: 1452–1456.

93. SaundersMA, LiangH, LiWH (2007) Human polymorphism at microRNAs and microRNA target sites. Proc Natl Acad Sci U S A 104: 3300–3305.

94. WangY, ShenD, BoS, ChenH, ZhengJ, et al. (2010) Sequence variation and selection of small RNAs in domesticated rice. BMC Evol Biol 10: 119.

95. TodescoM, BalasubramanianS, CaoJ, OttF, SureshkumarS, et al. (2012) Natural variation in biogenesis efficiency of individual Arabidopsis thaliana microRNAs. Curr Biol 22: 166–170.

96. WeiF, WingRA, WiseRP (2002) Genome dynamics and evolution of the Mla (powdery mildew) resistance locus in barley. Plant Cell 14: 1903–1917.

97. ShirasuK, Schulze-LefertP (2000) Regulators of cell death in disease resistance. Plant Mol Biol 44: 371–385.

98. HoltBF, BelkhadirY, DanglJL (2005) Antagonistic control of disease resistance protein stability in the plant immune system. Science 309: 929–932.

99. Schulze-LefertP, VogelJ (2000) Closing the ranks to attack by powdery mildew. Trends Plant Sci 5: 343–348.

100. Stirnweis D, Milani SD, Brunner S, Herren G, Buchmann G, et al. (2014) Suppression among alleles encoding nucleotide-binding–leucine-rich repeat resistance proteins interferes with resistance in F1 hybrid and allele-pyramided wheat plants. Plant J: doi:10.1111/tpj.12592. [Epub ahead of print].

101. Hurni S, Brunner S, Stirnweis D, Herren G, Peditto D, et al. (2014) The powdery mildew resistance gene Pm8 derived from rye is suppressed by its wheat ortholog Pm3. Plant J: doi:10.1111/tpj.12592. [Epub ahead of print].

102. NarusakaM, KuboY, HatakeyamaK, ImamuraJ, EzuraH, et al. (2013) Interfamily transfer of dual NB-LRR genes confers resistance to multiple pathogens. PLoS One 8: e55954.

103. ZukerM (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31: 3406–3415.

104. DongY, Burch-SmithTM, LiuY, MamillapalliP, Dinesh-KumarSP (2007) A ligation-independent cloning tobacco rattle virus vector for high-throughput virus-induced gene silencing identifies roles for NbMADS4-1 and -2 in floral development. Plant Physiol 145: 1161–1170.

105. ZhengL, BaumannU, ReymondJL (2004) An efficient one-step site-directed and site-saturation mutagenesis protocol. Nucleic Acids Res 32: e115.

106. ChenC, RidzonDA, BroomerAJ, ZhouZ, LeeDH, et al. (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33: e179.

107. LiuB, LiP, LiX, LiuC, CaoS, et al. (2005) Loss of function of OsDCL1 affects microRNA accumulation and causes developmental defects in rice. Plant Physiol 139: 296–305.

108. PallGS, HamiltonAJ (2008) Improved northern blot method for enhanced detection of small RNA. Nat Protoc 3: 1077–1084.

109. Van OoijenG, MayrG, KasiemMM, AlbrechtM, CornelissenBJ, et al. (2008) Structure-function analysis of the NB-ARC domain of plant disease resistance proteins. J Exp Bot 59: 1383–1397.

110. LaemmliUK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

111. LiuY, SchiffM, MaratheR, Dinesh-KumarSP (2002) Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 30: 415–429.

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2014 Číslo 12
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#