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Abstract
The ATAF1 protein is a member of the NAC transcription factor family and plays an important regulatory role in plant development and stress response. In this study, using rapid amplification of cDNA ends, a full-length cDNA of ATAF1 homolog was cloned from Populus trichocarpa, and the corresponding gene was named PtATAF1-1 . The full-length cDNA sequence of PtATAF1-1 is 1 242 bp, which consists of 3 exons and 2 introns, and an 873 bp coding region. The multiple amino acid sequences alignment showed that PtATAF1-1 protein contains the conserved NAM domain of NAC family. Transient expressing with poplar protoplast shows PtATAF1-1 protein localization in the nucleus. In addition, with the treatment of N-terminal conserved region of bacterial flagellin (Flg22) and abscisic acid (ABA) altered the expression level of PtATAF1-1 gene. In this study, the homologous cloning and functional analysis of the PtATAF1-1 gene were performed, which provided a reference for the further development of the function of ATAF1 gene in poplar .
Highlights
Populus is widely distributed in the middle latitudes of the world, and is one of the important industrial wood and ecological protection tree species in China (An, 2013, Modern Horticulture, (14): 69-71)
The results showed that in the case of single-transformed GFP, green fluorescence existed in the cell membrane and nucleus, while the green fluorescence occurred in the PtATAF1-1-GFP fusion protein only appeared in the nucleus, indicating that PtATAF1-1 protein was a nuclear localization protein, in line with the characteristics of its transcription factor (Figure 5). 1.5 Determination of PtATAF1-1 gene expression under abscisic acid (ABA) and Flg22 treatment The conserved regions of bacterial flagellin N-terminal Flg22 (2 μmol/L) and ABA (0.01 mmol/L) were used to treat tissue culture seedlings of Populus trichocarpa
It has been found that ATAF1 and ATAF2 genes in Arabidopsis thaliana are attacked and induced by pathogens (Zheng et al, 2009)
Summary
1.1 Cloning and sequence analysis of PtATAF1-1 homolog in Populus trichocarpa Blastp analysis of Arabidopsis AtATAF1 (AT1G01720) amino acid sequences in Populus trichocarpa database revealed 8 highly similar proteins, identified as Potri.002G081000, Potri.011G046700, Potri.011G123500, Potri.001G404100, Potri.005G180200, Potri.004G038000, Potri.011G123300, Potri.001G404400, E-Value is 1e-116, 6e-063, 2e-062, 2e-062, 6e-063, 7e-112, 3e-063, 5e-063. Blastp analysis was performed using these homologous proteins in The Arabidopsis Information Resource (TAIR), it was found that Potri.005G180200 and Potri.002G081000 were most likely to be ATAF1 homologous proteins in poplars They were named PtATAF1-1 and PtATAF1-2, respectively. 1.2 Evolutionary analysis of PtATAF1-1 protein In order to investigate the evolutionary relationship of PtATAF1-1 transcription factor, the amino acid sequences of ATAF1 homologous proteins in 16 species such as Populus trichocarpa, Hevea brasiliensis, Jatropha curcas and Salix brachista were used to construct phylogenetic trees. The phylogenetic trees were constructed by MEGA X software It turns out, ATAF1 of Populus trichocarpa and Salix brachista in Salicaceae, and Ricinus communis, Hevea brasiliensis, and Jatropha curca in Euphorbiaceae constituted a highly supported clade (94%), which was consistent with the close relationship between Salicaceae and Euphorbiaceae.
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