| Transcriptional regulation plays a critical role in the process of plant-pathogen interaction.Transcriptional factors(TFs),as functional proteins of transcriptional regulation,mediate plant disease resistance by activating or inhibiting the expression of resistance relatively genes.The conserved plant-specific ERF(Ethylene Responsive Factor)and BT(BTB and TAZ protein)play an important role in the development,abiotic stress and biotic stress.The reports which regarding the role of ERF TFs and BTs protein in the plant-biotrophic pathogen interaction rarely were reported.Therefore,this research screened and identified ERF TFs and BT proteins involved in Arabidopsis-Pseudomonas syringae interaction,and investigated the function of them in the plant resistance.Meanwhile,these results provided the importace theory for the breeding of varieties with improved resistance pathogen tolerance of crop.Main results in this paper were as follows:1.The AtERF11 was identified in the three absolute transcriptome datas about plant immune from GEO database.The expression of AtERF11 was induced by salicylic acid,Pseudomonas syringae pv.tomato(Pst)DC3000 infected transcriptome data,decreased in the sid2 related transcriptome data,suggesting that AtERF11 involve in SA regulated disease resistance pathways.The AtERF11 promoter contained some SA,JA,ABA,GA,ET,defense and stress responsiveness elements.The expression of AtERF11 was induced by SA,JA,ACC,and Pst DC3000.The related-induced was compromised in sid2,npr1-1,NahG,ein2,ein3 and ein3 eil1 plants.These indicate that AtERF11 is regulated by SA and ET signaling pathway.To compare with Col-0,the erf11 mutant exhibited a susceptible to Pst DC3000,whereas overexpression of ERF11 transgenic plants showed significant resistance phenotype.After Pst DC3000 infection,the expression of pathogen related genes PR1,PR2,PR5 in erf11 mutant is weakened,but expression of PR1,PR2,PR5 were intensified in the ERF11-OE transgenic lines.In Pst DC3000 infection,the content of ROS in the erf11 mutant and the content of AsA in the ERF11-OE plants were attenuated,but the content of ROS in the ERF11-OE plants and the content of AsA in the erf11 mutant were strengthened.Altered expression of AtERF11 affected the expression of ROS synthesis and AsA synthesis genes RBOHD,VTC1,and MIOX4.These indicate that AtERF11 play a positive role in the resistance to Pst DC3000.Loss function of AtERF11 affect SA-induced resistance and the expression of the pathogen related genes PR1,PR2,PR5.2.The characteristic analysis of AtERF11 showed that AtERF11 possessed essential characters of transcription factor,i.e nucleus location and transactivation activity.Transient assay and yeast one-hybird assay proved that AtERF11 interactd with GCC-box and DRE motif which were aslo specific binding sequnence of ERF transcription factor.The expression of AtBT4 was weakened in the erf11 mutant,and was strengthened in the ERF11-OE plants,suggesting that AtERF11 TFs mediate the expression of AtBT4.Tobacco transient assay confirmed that AtERF11 activated the expression of AtBT4.The yeast one-hybrid assay and EMSA proved the N-terminal of At ERF11 bound to the GCC-box site in-2065 bp from AtBT4 promoter.3.The expression of AtBT4 was induced by SA,JA,ACC,and Pst DC3000.The related-induced of AtBT4 were compromised in sid2,npr1-1,NahG,ein2,ein3 and ein3 eil1 plants.These indicated that At BT4 was regulated by SA,JA and ET signaling pathway.To compare with Col-0,the bt4-1 and bt4-2 mutants exhibited susceptible to Pst DC3000,whereas overexpression of BT4 transgenic plants showed significant resistance phenotype.After Pst DC3000 infection,the expression of pathogen related genes PR1,PR2,PR5 in bt4-1 and bt4-2 mutants were weakened,but expression of PR1,PR2,PR5 in the ERF11-OE transgenic lines was intensified.In Pst DC3000 infection,the content of ROS in the bt4-1 and bt4-2 mutants and the content of AsA in the BT4-OE plants were attenuated,but the content of ROS in the BT4-OE plants and the content of AsA in the bt4-1 and bt4-2 mutants were strengthened.Altered expression of AtBT4 affected the expression of AsA synthesis genes VTC1,and MIOX4.Loss function of AtBT4 affected SA-induced resistance and the expression of the pathogen related genes PR1,PR2,PR5. |
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