Identification of the ZTL and COP1 gene in Vitis vinifera and expression analysis of different light qualities and light transfers

Abstract: 【Objective】The ZTL and COP1 genes in Vitis vinifera were identified.Furthermore, the expression levels were detected after the plantlets were treated with different light conditions.【Methods】Fulllength cDNA sequences of ZTL and COP1 were successfully cloned from tissue cultural plantlets of the‘Red Globe’grape (Vitis vinifera) .To understand their structure and function, physical and chemical characterization, protein secondary structure, sub-cellular localization, secondary structure and phylogenetic relationships of the two genes above were determined by using bioinformatic analysis.The expressions of the two genes were analyzed by using qRT-PCR and applying different treatments.In addition, the fluorescence expressions of the grape test-tube plantlets were analyzed by using a fluorescence parameter and applying different treatments.【Results】The sequence analysis revealed that the open reading frame of ZTL was 1 239 bp in size, encoding 412 amino acids with ATG as the initiation codon and TAA astermination codon.COP1 showed that the open reading frame was 486 bp, encoding 160 amino acids with ATG as the initiation codon and TGA and TAA astermination codons.The results showed that ZTL exhibited the molecular formula of C1972H3107N553O576S20, which is a hydrophilic stable protein and also dis- played poor fat solubility.However, COP1 showed a molecular formula of C832H1291N209O225S9, which is a hydrophobic stable protein and showed favorable fat solublility.The protein of ZTL and COP1 showed the same secondary structure, which is comprised of alpha helix, bela turn, random coil and an extended strand.However, the ZTL showed random coil>extended strand>alpha helix>bela turn, the COP1showed that the extended strand>random coil>alpha helix>bela turn.Transmembrane domain and signal peptide showed that ZTL was the membrane protein and COP1 was the transmembrane protein.Subcellular localization analysis showed that the protein of the two genes was located in the cytoplasm.The functional domain showed that ZTL contains a highly conserved domain of F-box between the first to sixtieth amino acids.COP1 contains a highly conserved domain of WD40/YVTN between the 20th to 123rd amino acids.Homology analysis showed that the total similarity between ZTL and other plant homologues exhibited a 74.59%identity relationship in amino acid sequences.While the total similarity between COP1 and other plant homologues exhibited a 61.24%identity relationship in amino acid sequences.The phosphorylation site showed that ZTL consisted of 19 phosphorylation sites, including 9 serine phosphorylation sites, 9 threonine phosphorylation sites and 1 tyrosine phosphorylation site.COP1 consisted of 5phosphorylation sites, including 2 serine phosphorylation sites, 2 threonine phosphorylation sites and 1 tyrosine phosphorylation site.Phylogenetic analysis showed the most closely identified relationships between ZTL and the orthologous genes were in Prunus persica.While COP1 was much further from orthologous in the other species, which could be clustered into an individual subgroup.Compared with the control (white light) , the up-regulation of the ZTL gene were discovered within all treatments.Within them, the higher expressions were detected from white to red light, red light, red to white light and red to blue light treatments, which up-regulated 11.2, 22.3, 12.0, 32.9 and 19.6 folds, respectively.The different changes in COP1 expression were discovered from distinguishable light treatments.The expression was up-regulated 1.97 folds and treated by blue light with no significant changes in the red light treatment.Down-regulation was detected within other treatments.Within them, the expressions were down-regulated 18 folds from blue to white light and blue to red light treatments.The fluorescence parameter showed that the different changes in the q P expression were discovered from distinguishable light treatments.After red to blue, red, blue to red and red to white were significantly higher than the control (white light) , the expressions were up-regulated 1.97 folds treated by red to blue light.There was no significant difference between the expression level of the white light treatment and control (white) .The expression level was significantly lower than that in the control (white) after the blue treatment.The different changes in q N and NPQ expressions were discovered from distinguishable light treatments.After red to blue, red and blue to red were significantly higher than the control (white) , the expressions of q N and NPQ were up-regulated1.41 and 1.79 folds and treated by red to blue light.The expression level was significantly lower than that in the control (white) after the blue to white treatment.The different changes in Fv/Fm expression were discovered from distinguishable light treatments.After red to blue and blue to red were significantly higher than the control (white) , the expression of Fv/Fm was up-regulated 1.08 folds treated by red to blue light.There was no significant difference between the expression level of red, red to white, white to blue, blue and blue to white light treatment and the control (white) .The expression level was significantly lower than that in the control (white) after the white to red treatment.【Conclusion】Comprehensive analysis showed that the ZTL expression was active in the light of the grapes, and showed a significant upward trend, while COP1 showed a downward trend.Fluorescence parameter analysis showed that q P, q N, NPQ and Fv/Fm expressions were active in the red to blue light of the grapes, and showed a significant upward trend.