Abstract
Ethylene response factor (ERF) transcription factors play important roles in regulating immune responses in plants. In our study, we characterized a member of the ERF transcription factor family, VaERF20, from the Chinese wild Vitis genotype, V. amurensis Rupr “Shuangyou”. Phylogenetic analysis indicated that VaERF20 belongs to group IXc of the ERF family, in which many members are known to contribute to fighting pathogen infection. Consistent with this, expression of VaERF20 was induced by treatment with the necrotrophic fungal pathogen Botrytis cinerea (B. cinerea) in “Shuangyou” and V. vinifera “Red Globe”. Arabidopsis thaliana plants over-expressing VaERF20 displayed enhanced resistance to B. cinerea and the bacterium Pseudomonas syringae pv. tomato (Pst) DC3000. Patterns of pathogen-induced reactive oxygen species (ROS) accumulation were entirely distinct in B. cinerea and PstDC3000 inoculated plants. Examples of both salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) responsive defense genes were up-regulated after B. cinerea and PstDC3000 inoculation of the VaERF20-overexpressing transgenic A. thaliana plants. Evidence of pattern-triggered immunity (PTI), callose accumulation and stomatal defense, together with increased expression of PTI genes, was also greater in the transgenic lines. These data indicate that VaERF20 participates in various signal transduction pathways and acts as an inducer of immune responses.
Highlights
In order to protect themselves from a wide variety of pathogens, plants have evolved an innate immunity system, comprising two inter-connected components: pathogen/microbe-associated molecular pattern (PAMP or MAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) [1,2]
ETI is often accompanied by programmed cell death (PCD), called the hypersensitive response (HR), which reduces the spread of an invading pathogen [2]
The AP2/Ethylene response factor (ERF) gene family has previously been reported to be involved in the regulation of plant diseaTsheereAsiPst2a/nEcReFpgaethnwe afayms [il1y8,h29a,s42p]r.eIvniothuisslystbuedeyn, wreepcolrotnededtothbeeERinFv2o0lvgeedneinfrothme Crehgiunleasteiownilodf Vp.laanmtudreisnesaisseRurepsris“tSahnuceanpgaythowu”a,yasn[d18s,h2o9w,4e2d]. bIny tsheiqsusetnucdeya, nwaelycsliosntehdattthheeEpRroFt2e0inghenase tfhroemcoCnsheirnveesde EwRilFddVo.maaminusre(nSsuisppRluempren“tSahruyaDnagtyaoSu1”,anadndS2s)haonwdesdhabrye ssiemquileanricteiesanwailtyhsiitss tchoautnttheerpparrot tieninV.hvainsiftehrea “cPoinnsoetrvNeodirE”R
Summary
In order to protect themselves from a wide variety of pathogens, plants have evolved an innate immunity system, comprising two inter-connected components: pathogen/microbe-associated molecular pattern (PAMP or MAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) [1,2]. We cloned a grapevine ERF gene, VaERF20, which was chosen from the RNA-seq experiment, and over-expressed it in A. thaliana to examine its potential function in resistance to B. cinerea and PstDC3000. Expression levels of defense-related genes were assessed by qPCR in response to B. cinerea inoculation. The expression of the pathogenesis-related gene 1 (AtPR1) and lipoxygenase-3 (AtLOX3), which play major roles in the SA-dependent disease resistance response and JA biosynthesis, respectively, increased post infection (Figure 4G). WWhheenn tthhee DC3000 bacteria or the defense response elicitors flg and LPS were separately applied to the transgenic lines and Col-0, and the plants treated with the callose-binding stain aniline blue, we observed more callose in the transgenic lines than in WT (Figure 7). Asasmterpilsek.s iAnsdtiecraitseksstaitnisdtiiccaaltesigsntaifiticsatinccael (s*i*gnpi
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