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Transcriptome Profiling of the spl5 Mutant Reveals That SPL5 Has a Negative Role in the Biosynthesis of Serotonin for Rice Disease Resistance

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Transcriptome Profiling of the spl5 Mutant Reveals That SPL5 Has a Negative Role in the Biosynthesis of Serotonin for Rice Disease Resistance

Bin Jin et al. Rice (N Y).

Abstract

Background: Rice mutant, spl5 (spotted leaf 5), has spontaneous hypersensitive-like lesions on its leaves and shows enhanced resistance to pathogens, indicating that SPL5 plays a role in programmed cell death (PCD) and disease resistance. To understand the molecular mechanism of SPL5 gene, we investigated the transcriptome profiles of the spl5 mutant leaves with few lesions (FL) and leaves with many lesions (ML) compared to the wild-type (WT) leaves respectively by microarray.

Results: The data from microarray revealed that 243 and 896 candidate genes (Fold change ≥ 3.0) were up- or down-regulated in the spl5-FL and spl5-ML, respectively, and a large number of these genes involved in biotic defense responses or reactive oxygen species (ROS) metabolism. Interestingly, according to our microarray and real-time PCR assays, the expressions of a transcription factor OsWRKY14 and genes responsible for the biosynthesis of serotonin, anthranilate synthase (AS), indole-3-glycerolphosphate synthase (IGPS), tryptophan synthase (TS) and tryptophan decarboxylase (TDC) were significantly up-regulated in the spl5 mutant. It has been reported previously that TS and TDC expressions are regulated by OsWRKY14 in rice, which raises the possibility that OsWRKY14 regulates serotonin production through the up-regulation of TS and TDC. Our HPLC analysis further confirmed that serotonin levels were higher in the leaves of spl5 mutant than that in WT.

Conclusions: Since the serotonin plays a critical role in inducing disease-resistance, the increased serotonin level may contribute, at least partly, to the disease resistance in spl5. The SPL5 gene may act as a negative regulatory factor activating the serotonin metabolic pathway, and these results might provide a new insight into the spl5-induced defense response mechanisms in plants.

Keywords: Disease resistance; Lesion mimic; Microarray; Rice; Serotonin; spl5.

Figures

Figure 1
Figure 1
Expressions of OsWRKY14, AS, IGPS, TS and TDC by real-time PCR analysis. The OsWRKY14, AS (anthranilate synthase), IGPS (indole-3-glycerolphosphate synthase), TS (tryptophan synthase) and TDC (tryptophan decarboxylase) genes expressions are shown for the WT leaves and the spl5 leaves with different degrees of lesion development: NL (no lesions), FL (few lesions) and ML (many lesions). The accession number of gene is from the NCBI database (http://www.ncbi.nlm.nih.gov/). The significance of expression compared to WT with the P value less than 0.05 and 0.01 are marked by * and **, respectively.
Figure 2
Figure 2
In vivo tryptophan and serotonin levels by HPLC analysis. The tryptophan and serotonin concentrations are shown for the WT leaves and the spl5-NL, spl5-FL, and spl5-ML leaves, respectively. The significance of expression compared to WT with the P value less than 0.05 and 0.01 are marked by * and **, respectively.
Figure 3
Figure 3
SL and OsPR1a expressions by real-time PCR analysis. The SL and OsPR1a gene expressions are shown for the spl5 leaf parts with different lesion-mimic numbers and for WT. The significance of expression compared to WT with the P value less than 0.05 and 0.01 are marked by * and **, respectively.
Figure 4
Figure 4
A hypothetical model for the SPL5 signaling pathway in rice. The SPL5 may act as a negative regulatory factor activating the serotonin metabolic pathway, which is mediated by OsWRKY14. The accumulation of serotonin may lead to pathogen resistance in rice.

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