葡萄耐铝毒基因 STOP1 的克隆与表达分析.

[Objective] STOP1 is an important gene for tolerance to aluminum (Al) -toxicity in plants. The purpose of the present study was to explore the relationship between the structural characteristics, expression level of STOP1 gene and its tolerance to Al in grape. [Method] Leaf cDNA of Vitis amurensis with weak tolerance to Al and V. sinocinerea with strong tolerance to Al were used as reference materials. Sequences of two genes VaSTOP1 and VsSTOP1 were obtained by PCR and TA cloning technology, and then bioinformatics analysis and expression analysis under Al stress were carried out. The membrane lipid peroxidation indices under Al stress were determined. Finally, the relationship between the expression of STOP1 genes, the changes of membrane lipid peroxidation of two species and tolerance to Al were compared. [Result] The sequences of VaSTOP1 and VsSTOP1 were 1782 and 1800 bp respectively, encoding 594 and 600 amino acids. Their relative molecular weights were 67. 03 and 67. 48 kD, and the theoretical isoelectric points were 5. 21and6. 42, respectively. The secondary structure in two species were the law of irregular curl > α-helix > folding extension chain, and the tertiary spatial structure were composed of zinc protein, α-helix, β-folded sheet and β-rotation angle. The phylogenetic tree analysis found that the homology between VaSTOP1 and VsSTOP1 was as high as 96%, and the homology between them and CcSTOP1, PnSTOP1, NtSTOP1 and ErSTOP1 was more than 80%. The expression of VaSTOP1 gene increased first and then decreased within 28 days under Al stress, but the peak value appeared earlier, whereas the VsSTOP1 gene expression increased steadily, which was basically consistent with the change law of membrane lipid peroxidation. [Conclusion] The tolerance to Al of Vitis was positively correlated with the expression of STOP1 gene, which provided a reference for further exploring the molecular regulation mechanism of Vitis tolerance to Al. [ABSTRACT FROM AUTHOR]