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以抗性葡萄品种‘F-242’组培苗为材料,利用同源克隆法克隆了葡萄VvBAP1基因。测序结果显示,VvBAP1扩增片段大小为531 bp,可编码176个氨基酸序列。利用生物信息学分析VvBAP1基因编码的蛋白序列显示,该蛋白分子量为19.43kDa,含有保守的钙离子依赖性的C2结构域;等电点pI为9.42;不稳定系数为37.09,推测为稳定的亲水性蛋白;含有多个丝氨酸/苏氨酸磷酸化位点。实时荧光定量PCR表明,该基因在根茎叶中均有表达,其中在叶片中表达量较高;盐胁迫、低温等逆境因子及逆境相关的信号物质,如水杨酸和一氧化氮均可诱导VvBAP1的表达,其中低温对其表达量影响更为显著,推测该基因参与了葡萄抵御逆境胁迫的过程,尤其是与低温相关的过程。
The resistant grape variety ’F-242’ was used to clone the VvBAP1 gene of grapevine by homologous cloning method. The sequencing results showed that the size of the amplified fragment of VvBAP1 was 531 bp, which encoded a 176 amino acid sequence. Bioinformatics analysis of the protein sequence encoded by the VvBAP1 gene showed that the protein had a molecular weight of 19.43 kDa and contained a conserved calcium-dependent C2 domain with an isoelectric point of 9.42 and an instability coefficient of 37.09, suggesting a stable pro- Aqueous protein; contains multiple serine / threonine phosphorylation sites. Real-time quantitative PCR showed that the gene was expressed in rhizomes and leaves, and the expression level was high in leaves. The stress factors such as salt stress, low temperature and other adverse signals such as salicylic acid and nitric oxide could induce VvBAP1 , Of which low temperature had a more significant impact on its expression level. It is speculated that the gene is involved in the process of grape resistance to stress, especially in the process of low temperature.