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. 2016 Mar 3;11(3):e0150711.
doi: 10.1371/journal.pone.0150711. eCollection 2016.

Insight on Genes Affecting Tuber Development in Potato upon Potato spindle tuber viroid (PSTVd) Infection

Konstantina Katsarou  1 , Yun Wu  2 , Runxuan Zhang  3 , Nicola Bonar  4 , Jenny Morris  4 , Pete E Hedley  4 , Glenn J Bryan  4 , Kriton Kalantidis  1   5 , Csaba Hornyik  4
Affiliations

Insight on Genes Affecting Tuber Development in Potato upon Potato spindle tuber viroid (PSTVd) Infection

Konstantina Katsarou et al. PLoS One. .

Abstract

Potato (Solanum tuberosum L) is a natural host of Potato spindle tuber viroid (PSTVd) which can cause characteristic symptoms on developing plants including stunting phenotype and distortion of leaves and tubers. PSTVd is the type species of the family Pospiviroidae, and can replicate in the nucleus and move systemically throughout the plant. It is not well understood how the viroid can affect host genes for successful invasion and which genes show altered expression levels upon infection. Our primary focus in this study is the identification of genes which can affect tuber formation since viroid infection can strongly influence tuber development and especially tuber shape. In this study, we used a large-scale method to identify differentially expressed genes in potato. We have identified defence, stress and sugar metabolism related genes having altered expression levels upon infection. Additionally, hormone pathway related genes showed significant up- or down-regulation. DWARF1/DIMINUTO, Gibberellin 7-oxidase and BEL5 transcripts were identified and validated showing differential expression in viroid infected tissues. Our study suggests that gibberellin and brassinosteroid pathways have a possible role in tuber development upon PSTVd infection.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Symptom development and PSTVd accumulation in infected potato plants.
(A) Mock inoculated Safari plant (right) and PSTVd infected plant (left) at 14 weeks post-inoculation (wpi). (B) Tubers of mock inoculated (WT) and infected (INF) plants at 14 wpi on the upper panels and 21 wpi on the lower panels. Numbers indicate the different individual plants. White arrows show which tubers were used for RNA extraction. (C) Northern blot analysis of mock inoculated (WT) and infected (INF) plants. Numbers indicate different individual plants. Upper panel shows viroid accumulation in early (14 wpi) samples and lower panel shows the presence of PSTVd in late (21 wpi) samples. Methylene blue staining of ribosomal RNAs was used as loading control; the arrow shows the 25S ribosomal RNA which was used for normalization. The quantification of viroid signal is shown as numbers under the viroid RNA gel blot panels. Relative normalisation was performed for each plant; normalised to leaf viroid level using ribosomal RNA as control.
Fig 2
Fig 2. Heatmap representation of differentially expressed genes upon PSTVd infection.
Visualization of differentially expressed genes of leaf (A) and tuber (B) tissues in mock inoculated and infected plants as indicated. Three biological replicates are shown, but for late infected tissues only two. Colour key indicates the expression change compared to mock inoculated samples.
Fig 3
Fig 3. Representation of GO term enrichment.
Selected GO term enrichments are visualized in early (A) and late (B) leaf samples. Figs (C) and (D) show GO term enrichment for early and late tuber samples, respectively. Categories are indicated on Y axis, log10[1/p] is on the X-axis where p is the p-value of the enrichment.
Fig 4
Fig 4. Quantitative RT-PCR analysis of candidate genes.
Mock infected plants were used to normalize fold change of early and late samples in leaf and tuber tissues as indicated. White bars represent the RT-qPCR analysis; grey bars show the microarray data. Error bars represent standard error (SE). L23 and PP2A genes were used as internal normalization controls. Asterisk indicates that the means of fold change of the mock and infected samples are significantly different (Student’s t-test).
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Grants and funding

This work was supported by the Scottish Government Rural and Environment Science and Analytical Services Division (RESAS). YW spent 15 months at The James Hutton Institute (Invergowrie, Scotland, UK) funded by China Scholarship Council (201306320084). KoK was partially funded by the Greek ministry for education and religious affairs (program Aristeia II, 4499, ViroidmiR; ESPA 2007-2013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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