Abstract
Key message
Cowpea miRNAs and Argonaute genes showed differential expression patterns in response to CPSMV challenge
Abstract
Several biotic stresses affect cowpea production and yield. CPSMV stands out for causing severe negative impacts on cowpea. Plants have two main induced immune systems. In the basal system (PTI, PAMP-triggered immunity), plants recognize and respond to conserved molecular patterns associated with pathogens (PAMPs). The second type (ETI, Effector-triggered immunity) is induced after plant recognition of specific factors from pathogens. RNA silencing is another important defense mechanism in plants. Our research group has been using biochemical and proteomic approaches to learn which proteins and pathways are involved and could explain why some cowpea genotypes are resistant whereas others are susceptible to CPSMV. This current study was conducted to determine the role of cowpea miRNA in the interaction between a resistant cowpea genotype (BRS-Marataoã) and CPSMV. Previously identified and deposited plant microRNA sequences were used to find out all possible microRNAs in the cowpea genome. This search detected 617 mature microRNAs, which were distributed in 89 microRNA families. Next, 4 out of these 617 miRNAs and their possible target genes that encode the proteins Kat-p80, DEAD-Box, GST, and SPB9, all involved in the defense response of cowpea to CPSMV, had their expression compared between cowpea leaves uninoculated and inoculated with CPSMV. Additionally, the differential expression of genes that encode the Argonaute (AGO) proteins 1, 2, 4, 6, and 10 is reported. In summary, the studied miRNAs and AGO 2 and AGO4 associated genes showed differential expression patterns in response to CPSMV challenge, which indicate their role in cowpea defense.
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This work was financially supported by the Council for Advanced Professional Training (CAPES), the National Council for Scientific and Technological Development (CNPq, grant numbers 431511/2016-0 and 306202/2017-4 to JTAO, and Project Finance Code 431511/2016-0 to IMV). CAPES sponsored TFM with a doctoral grant.
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Fig. S1 Average expression stability of insulinase, fosfolipase A2, protein L23A1, L23A3, and PhaF-box genes using cowpea cDNA from leaves mock-inoculated and inoculated with CPSMV (a). Average expression stability was determined by GeNorm. More stably expressed genes are positioned on the right side of the diagram and the less stably expressed on the right side. Determination of the optimal number of V. unguiculata reference genes for qRT-PCR data normalization by pairwise variation (geNorm V) was done using the GeNorm algorithm (b). A pairwise variation (Vn/Vn+1) below 0.15 indicates that the optimum number of reference gene for qRT-PCR data normalization is n.
Fig. S2 Relative expression levels (qRT-PCR) of vun-miR156 (a) and its respective target gene SPB9 (b) in leaves of BRS-Marataoã inoculated with CPSMV. Expression levels were calculated at 0, 1, 2, and 6 DAI in relation to reference genes. Comparison of the transcript and gene expression levels was done between cowpea plants inoculated with CPSMV in relation to the respective mock-inoculated control.
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Martins, T.F., Souza, P.F.N., Alves, M.S. et al. Identification, characterization, and expression analysis of cowpea (Vigna unguiculata [L.] Walp.) miRNAs in response to cowpea severe mosaic virus (CPSMV) challenge. Plant Cell Rep 39, 1061–1078 (2020). https://doi.org/10.1007/s00299-020-02548-6
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DOI: https://doi.org/10.1007/s00299-020-02548-6