Abstract:
To explore the responses of physiological and biochemical and mechanism of
molecular regulatory for grape dormancy breaking with cyanamide, Vitis vinifera×V. labrusca
‘Shuijing’ was used as experimental materials in this study to determine the changes in SOD, POD,
CAT activity, MDA and H2O2 content, and oxygen free radical production rate, and RT-PCR technology was used to clone the full-length cDNA sequences of two FT (Flowering location T)
genes (VvFT1 and VvFT2) and one CBF (C-repeat binding factor) gene (VvCBF) from its buds,
then their physicochemical properties, phylogenetic evolution, conserved motifs and domains, and
differences expression levels in grape buds after cyanamide treatment were analyzed. The results
were as follows: (1) The analysis of physiological and biochemical indicators results showed that
SOD, POD and CAT activities, MDA and H2O2 contents, and the rate of oxygen free radical
production in grape buds were significantly increased after treated with cyanamide. (2) The
full-lengths cDNA sequences of VvFT1 and VvFT2 genes were 525 bp from Vitis vinifera×V.
labrusca ‘Shuijing’, encoding 174 aa, and the full-lengths cDNA sequences of VvCBF gene was
714 bp, encoding 237 aa. (3) The homology analysis results showed that VvFT1 of Vitis
vinifera×V. labrusca ‘Shuijing’ had the highest amino acid homology with Litchi chinensis (LcFT:
AEU08960.1) and Dimocarpus longan (DlFT2: ALA55998.1), VvFT2 had the highest amino acid
homology with LcFT (AEU08961.1) and DlFT2 (AHF27444.1). The phylogenetic analysis result
showed that VvFT1, VvFT2, LcFT (AEU08960.1; AEU08961.1) and DlFT2 (ALA55998.1;
AHF27444.1) clustered into a branch, with the most closes genetic relationship; VvCBF had the
highest amino acid homology with Prunus ledebouriana (PlCBF: AEB69782.1), and the
phylogenetic analysis showed that VvCBF and PlCBF clustered into a branch, with the most
closest genetic relationship. (4) qRT-PCR analysis showed that VvFT1 and VvFT2 expression
levels were significantly increased in buds after treated with cyanamide, while VvCBF expression
levels was significantly decreased. In summary, this study comprehensively analyzed the
phylogenetic evolution of VvFT1, VvFT2, and VvCBF genes, as well as the expression patterns of
these genes and physiological and biochemical indicators in grape buds after treated with
cyanamide, laying a theoretical foundation for the molecular and physiological mechanisms of
grape dormancy breaking with cyanamide.
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