A Proteomics Study On The Effects Of Transcription Factor LeERF2 On Enhancing The Sensitivity To Abscisic Acid In Arabidopsis Thaliana

ERFs(Etylene Responsive Factors) play the significant regulatory role in the plants growth, development and stress response. LeERF2, as a new member of ERFs transcription factor, which has 780bp coding domain, encoding 260 amino acids, was isolated from tomato expression cDNA library using Gcc-box as bait by the yeast one hybrid which posseses typical characterization of transcription factor. In previous study, LeERF2 was able to enhance abscisic acid sensitivity during seedling development. However, it is no clear understood that how the gene regulators ABA signaling response network.Two-dimensional electrophoresis was used to constrct the differentially expressed proteins maps of wildtype and transgenic LeERF2 Arabidopsis thaliana cultivated in 1/2 MS agar plates or 1/2 MS agar plates supplenmented with ABA. Important differentially expressed proteins identified by mass spectrometry provide basis for future study of molecular merchanism of LeERF2 enhancing the sensitivity to abscisic acid in Arabidopsis thaliana by bioinformatics.Main Results summerized as follows:(1)Two-dimensional electrophoresis based on immobiled pH gradient tenchnique was used to establish differentially expressed proteins patterns owing to effect of LeERF2 on ABA response pathway by analyzing of WT and ecotopic LeERF2 Arabidopsis thaliana grown in 1/2 MS medium which contains ABA phytohormone or not(2) Through gels analysis and mass spectrometry identification,50 proteins were expressedly changed by qualitative or quantitative method in WT vs OE analysis group.10 Of them was founded in wildtype, and 2 was detected in transgenic seedlings; analysis of WT vs WTABA indicated that 49 proteins were observed.9 and 13 of them was differentially expressed in WT and WTABA, respectively; the total number of differentially expressed proteins was 49, and up-regulated and down-regulated account for 71.4% of change proteins when wildtype and transgenic LeERF2 Arabidopsis thaliana both treated with 0.1μM ABA.36 differentially expressed proteins were investigated by analzing transgenic LeERF2 plants before and after under 0.1μM ABA treatment in proein lever change.5 and 13 proteins showed only in OE and OEABA palnt, respectively.(3)155 proteins regulated by LeERF2 of wildtype and transgenic Arabidopsis thaliana plated on 1/2 MS agar medium or 1/2 MS agar medium including ABA which were regulated by LeERF2 such as ATI G31180 3-isopropylmalate dehydrogenase 1,AT1G52690 late embryogenesis abundant protein, putative,XYL1 Alpha-xylosidase,ATMS1 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase were identified.(4)These low concentration ABA-responsive proteins in wildtype Arabidopsis thaliana:including ATPB ATP synthase subunit beta,AT3G04790 ribose 5-phosphate isomerase-related,AT3G63170 chalcone isomerase, etc were identified.(5)Such as ATPQ ATP synthase subunit d,AT3G52960 Peroxiredoxin-2E,CB5-E Cytochrome b5 isoform 1 is regulated by LeERF2 in normal condition and differentially expressed proteins such as AT5G02490 Heat shock cognate 70 kDa protein 2,GAPA Glyceraldehyde-3-phosphate dehydrogenase A,CA1 Isoform 3 of Carbonic anhydrase were regulated by LeERF2 under 0.1μM ABA.Other differentially expressed proteins remain to be identifified.