Transcriptome analysis provides insights into the delayed sticky disease symptoms in Carica papaya

  • Johana Madroñero
  • Silas P. Rodrigues
  • Tathiana F. S. Antunes
  • Paolla M. V. Abreu
  • José A. Ventura
  • A. Alberto R. Fernandes
  • Patricia Machado Bueno Fernandes
Original Article

Abstract

Key message

Global gene expression analysis indicates host stress responses, mainly those mediated by SA, associated to the tolerance to sticky disease symptoms at pre-flowering stage in Carica papaya.

Abstract

Carica papaya plants develop the papaya sticky disease (PSD) as a result of the combined infection of papaya meleira virus (PMeV) and papaya meleira virus 2 (PMeV2), or PMeV complex. PSD symptoms appear only after C. papaya flowers. To understand the mechanisms involved in this phenomenon, the global gene expression patterns of PMeV complex-infected C. papaya at pre-and post-flowering stages were assessed by RNA-Seq. The result was 633 and 88 differentially expressed genes at pre- and post-flowering stages, respectively. At pre-flowering stage, genes related to stress and transport were up-regulated while metabolism-related genes were down-regulated. It was observed that induction of several salicylic acid (SA)-activated genes, including PR1, PR2, PR5, WRKY transcription factors, ROS and callose genes, suggesting SA signaling involvement in the delayed symptoms. In fact, pre-flowering C. papaya treated with exogenous SA showed a tendency to decrease the PMeV and PMeV2 loads when compared to control plants. However, pre-flowering C. papaya also accumulated transcripts encoding a NPR1-inhibitor (NPR1-I/NIM1-I) candidate, genes coding for UDP-glucosyltransferases (UGTs) and several genes involved with ethylene pathway, known to be negative regulators of SA signaling. At post-flowering, when PSD symptoms appeared, the down-regulation of PR-1 encoding gene and the induction of BSMT1 and JA metabolism-related genes were observed. Hence, SA signaling likely operates at the pre-flowering stage of PMeV complex-infected C. papaya inhibiting the development of PSD symptoms, but the induction of its negative regulators prevents the full-scale and long-lasting tolerance.

Keywords

Carica papaya Papaya meleira virus Transcriptome Plant–virus interaction Defense responses SA signaling 

Notes

Acknowledgements

This work was supported by the Fundação de Amparo à Pesquisa do Estado do Espírito Santo, FAPES, Grants #48497231 and #59899549/12. A.A.R. Fernandes, P.M.B. Fernandes and José A. Ventura were granted with the research productivity award from the Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, Grant #303902/2013-2, #304719/2014-5 and #307752/2012-7, respectively. L.J. Madroñero, T.F.S. Antunes and P.M.V. Abreu acknowledge the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES, for their schollarships.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

299_2018_2281_MOESM1_ESM.tif (37 kb)
Fig. S1 Field experimental time line. One month post germination (MPG), C. papaya seedlings (n=6) were transferred to an experimental field. Two months later, plants (n=3) were either injected in the stem apex with a suspension (1:1, v/v) of latex collected from sticky-diseased papaya fruits in 50 mM sodium phosphate buffer, pH 7.0 (treatment), or injected with phosphate buffer (control). The plants remained in the field until floral buds formed (at 4 MPG) (TIF 36 KB)
299_2018_2281_MOESM2_ESM.tif (95 kb)
Fig. S2 Venn diagram representing the relationship between all transcripts present in all samples. The reads mapped to 19,518 transcripts from 27,796 annotated transcripts for C. papaya. There were 18,392 in common in treatment (PMeV complex-infected plants) and control (uninfected plants) conditions at both times. There were 525 and 601 transcripts found only at pre-flowering (195 in treatment, 224 in control and 106 in both) and at post-flowering (269 in treatment, 188 in control and 144 in both), respectively (TIF 94 KB)
299_2018_2281_MOESM3_ESM.tif (442 kb)
Fig. S3 Protein sequence alignment of cpNPR1-I, jcNIM1-I and rNRR. C. papaya NPR1 interacting protein (cpNPR1-I, evm.TU.supercontig _1096.2), Jatropha curcas NIM1 interacting protein (jcNIM1-I) and the rice negative regulator of resistance (rNRR) were aligned using CLUSTAW. Alignmenet gaps are filled with hifens. Identical aminoacids are shaded. Putative nuclear localization motifs are in bold/underlined. Putative ERF-associated amphiphilic repression (EAR) motifs are in bold/underlined and shaded (TIF 442 KB)
299_2018_2281_MOESM4_ESM.tif (64 kb)
Fig. S4 Correlation between C. papaya mRNA abundance levels obtained by RNA-seq and qRT-PCR. The log2 ratios of PMeV complex-infected/control C. papaya mRNA abundances were compared for twenty-two genes at pre-flowering and nine genes at post-flowering (Supplementary Table S9) (TIF 64 KB)
299_2018_2281_MOESM5_ESM.tif (861 kb)
Fig. S5 Abundance levels of PMeV and PMeV2 in C. papaya plants after inoculation. Viral load of PMeV and PMeV2 were assessed after inoculation in pre-flowering (solid bars) and post-flowering (patterned bars) plants. Data shown are mean of Log10 number of cDNA viral copies ± SE from three biological replicates. p-values shown in the figure were obtained using t test (TIF 860 KB)
299_2018_2281_MOESM6_ESM.xls (6.8 mb)
Supplementary material 6 (XLS 7003 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Núcleo de BiotecnologiaUniversidade Federal do Espírito SantoVitóriaBrazil
  2. 2.Núcleo Multidisciplinar de Pesquisa-Polo de XerémUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  3. 3.Instituto Capixaba de PesquisaAssistência Técnica e Extensão RuralVitóriaBrazil

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