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      Diterpenoid compounds from Wedelia trilobata induce resistance to Tomato spotted wilt virus via the JA signal pathway in tobacco plants

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          Abstract

          Tomato spotted wilt virus (TSWV) causes major losses of many crops worldwide. Several strategies have been attempted to control disease caused by TSWV. However, many challenges for the effective control of this disease remain. A promising approach is the use of abiotic or biotic inducers to enhance plant resistance to pathogens. We screened a diterpenoid compound from Wedelia trilobata, 3α-Angeloyloxy-9β-hydroxy- ent-kaur-16-en-19-oic acid (AHK), which had higher curative and protective effects against TSWV than the ningnanmycin control. The rapid initiation of the expression of all the TSWV genes was delayed by more than 1d in the curative assay, and the expression of the NSs, NSm and RdRp genes was inhibited. In addition, the replication of all TSWV genes in systemic leaves was inhibited in the protective assay, with an inhibition rate of more than 90%. The concentrations of jasmonic acid (JA) and jasmonic acid isoleucine (JA-ILE) in the AHK-treated and systemic leaves of the treated plants were significantly higher than those observed in the control. The results suggested that AHK can induce systemic resistance in treated plants. The transcription of the NtCOI1 gene, a key gene in the JA pathway, was significantly higher in both the inoculated and systemic leaves of the AHK-treated plants compared to the control. The AHK-induced resistance to TSWV in Nicotiana benthamiana could be eliminated by VIGS-mediated silencing of the NtCOI1 gene. These results indicated that AHK can activate the JA pathway and induce systemic resistance to TSWV infection.

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          Most cited references42

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          COI1: an Arabidopsis gene required for jasmonate-regulated defense and fertility.

          The coi1 mutation defines an Arabidopsis gene required for response to jasmonates, which regulate defense against insects and pathogens, wound healing, and pollen fertility. The wild-type allele, COI1, was mapped to a 90-kilobase genomic fragment and located by complementation of coi1-1 mutants. The predicted amino acid sequence of the COI1 protein contains 16 leucine-rich repeats and an F-box motif. It has similarity to the F-box proteins Arabidopsis TIR1, human Skp2, and yeast Grr1, which appear to function by targeting repressor proteins for removal by ubiquitination.
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            Technical Advance. Tobacco rattle virus as a vector for analysis of gene function by silencing.

            Virus vectors carrying host-derived sequence inserts induce silencing of the corresponding genes in infected plants. This virus-induced gene silencing (VIGS) is a manifestation of an RNA-mediated defence mechanism that is related to post-transcriptional gene silencing (PTGS) in transgenic plants. Here we describe an infectious cDNA clone of tobacco rattle virus (TRV) that has been modified to facilitate insertion of non-viral sequence and subsequent infection to plants. We show that this vector mediates VIGS of endogenous genes in the absence of virus-induced symptoms. Unlike other RNA virus vectors that have been used previously for VIGS, the TRV construct is able to target host RNAs in the growing points of plants. These features indicate that the TRV vector will have wide application for gene discovery in plants.
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              The SCF(COI1) ubiquitin-ligase complexes are required for jasmonate response in Arabidopsis.

              Xie and colleagues previously isolated the Arabidopsis COI1 gene that is required for response to jasmonates (JAs), which regulate root growth, pollen fertility, wound healing, and defense against insects and pathogens. In this study, we demonstrate that COI1 associates physically with AtCUL1, AtRbx1, and either of the Arabidopsis Skp1-like proteins ASK1 or ASK2 to assemble ubiquitin-ligase complexes, which we have designated SCF(COI1). COI1(E22A), a single amino acid substitution in the F-box motif of COI1, abolishes the formation of the SCF(COI1) complexes and results in loss of the JA response. AtRbx1 double-stranded RNA-mediated genetic interference reduces AtRbx1 expression and affects JA-inducible gene expression. Furthermore, we show that the AtCUL1 component of SCF(COI1) complexes is modified in planta, where mutations in AXR1 decrease the abundance of the modified AtCUL1 of SCF(COI1) and lead to a reduction in JA response. Finally, we demonstrate that the axr1 and coi1 mutations display a synergistic genetic interaction in the double mutant. These results suggest that the COI1-mediated JA response is dependent on the SCF(COI1) complexes in Arabidopsis and that the AXR1-dependent modification of the AtCUL1 subunit of SCF(COI1) complexes is important for JA signaling.
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                Author and article information

                Contributors
                zhongkai99@sina.com
                dongjhn@126.com
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                26 February 2019
                26 February 2019
                2019
                : 9
                : 2763
                Affiliations
                [1 ]ISNI 0000 0004 1799 1111, GRID grid.410732.3, Institute of Biotechnology and Germplasm Resources, , Yunnan Academy of Agricultural Sciences, Yunnan Provincial Key Laboratory of Agricultural Biotechnology, Key Lab of Southwestern Crop Gene Resource and Germplasm Innovation, Ministry of Agriculture, ; 650204 Kunming, China
                [2 ]ISNI 0000000119573309, GRID grid.9227.e, Kunming Institute of Botany, , Chinese Academy of Science, ; 650201 Kunming, China
                [3 ]ISNI 0000 0004 1759 700X, GRID grid.13402.34, Institute of Biotechnology, , Zhejiang University, ; Hangzhou, 310058 Zhejiang China
                [4 ]ISNI 0000 0004 1761 2943, GRID grid.412720.2, Life Science College, , Southwest Forestry University, ; 650224 Kunming, China
                Article
                39247
                10.1038/s41598-019-39247-6
                6391457
                30808959
                5d5e62d6-6eda-4177-83b1-8856ce7e01b7
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 23 January 2018
                : 8 January 2019
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001809, National Natural Science Foundation of China (National Science Foundation of China);
                Award ID: 31470427; 31371919
                Award Recipient :
                Funded by: Yunnan Provincial Top Scientists Input Program(2013HA028)
                Funded by: FundRef https://doi.org/10.13039/501100005273, Natural Science Foundation of Yunnan Province (Yunnan Provincial Natural Science Foundation);
                Award ID: 2017FA019; 2012CH007
                Award Recipient :
                Funded by: Yunling Scholar Program (2015zhangzhongkai)
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