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Ethylene response factors (ERFs) play an important role in plants by regulating defense response through interaction with various stress pathways. After harvest, table grapes ( Vitis vinifera  L.) are subject to a range of problems associated with postharvest storage at 0°C, such as fungal attack, water loss and rachis browning. The application of a 3-day high CO 2  treatment maintained fruit quality and activated the induction of transcription factors belonging to different families such as ERF. In this paper, we have isolated five  VviERFs  from table grapes cv. Cardinal, whose deduced amino acid sequence contained the conserved apetalous (AP2)/ERF domain. The phylogeny and putative conserved motifs in VviERFs were analyzed and compared with those previously reported in  Vitis .  VviERFs-c  gene expression was studied by quantitative real-time RT-PCR in the different tissues of bunches stored at low temperature and treated with high levels of CO 2 . The results showed that in most of the tissues analyzed,  VviERFs-c  gene expression was induced by the storage under normal atmosphere although the application of high levels of CO 2  caused a greater increase in the  VviERFs-c  transcript accumulation. The promoter regions of two PRs (pathogenesis related proteins),  Vcchit1b  and  Vcgns1 , were obtained and the  in silico  analysis revealed the presence of a  cis -acting ethylene response element (GCC box). In addition, expression of these two PR genes was analyzed in the pulp and rachis of CO 2 -treated and non-treated table grapes stored at 0°C and results showed significant correlations with  VviERF2-c  and  VviERF6L7-c  gene expression in rachis, and between  VviERF11-c  and  Vcchit1b  in pulp. Finally by using electro mobility shift assays, we denoted differences in binding of VviERFs to the GCC sequences present in the promoters of both PRs, with VviERF6L7-c being the only member which did not bind to any tested probe. Overall, our results suggest that the beneficial effect of high CO 2  treatment maintaining table grape quality seems to be mediated by the regulation of  ERFs  and in particular  VviERF2-c  might play an important role by modulating the expression of PR genes.

frontiers in plant science

Expression Profiles and DNA-Binding Affinity of Five ERF Genes in Bunches of Vitis vinifera cv. Cardinal Treated with High Levels of CO2 at Low Temperature