中文摘要：

WRKY转录因子是植物特有的一类超家族转录因子,在植物的生长、发育和衰老过程中都有WRKY转录因子的参与。本研究利用SRAP-cDNA技术,以无核葡萄（Vitis vinifera）品种无核白胚珠各发育时期的cDNA为材料,克隆获得了311bp的差异表达片段,进一步在有核葡萄（V. vinifera）品种黑比诺胚珠中进行验证。BLASTn分析表明,差异表达片段为葡萄基因组预测的WRKY20基因序列（XM_002272334.2）的部分序列。以序列XM_002272334.2为模版设计引物,通过RT-PCR技术在无核白胚珠cDNA中克隆到一个长1796bp的同源序列,其ORF为1653bp,编码550个氨基酸,命名为VvWRKY20（GenBank登录号：JQ782602）。生物信息学分析表明,VvWRKY20含两个WRKY保守结构域,两个C2H2锌指结构域,两个DNA结合位点,为WRKY家族中的Ⅰ类成员。其分子量为60140.4D,等电点为6.10,不稳定系数为51.01,属于不稳定蛋白。实时荧光定量PCR分析,该基因在无核白胚珠发育过程中表达量大且变化十分很明显,最大值与最小值之比为17.56,在有核品种黑比诺中,表达较低且表达水平保持相对稳定,最大值与最小值之比仅为2.52。在同一发育时期,该基因在无核白中的表达量也远高于在黑比诺葡萄中的表达量,10d时为15.77倍,30d时达到229.12倍。有核与无核品种VvWRKY20基因表达的差异表明,该基因可能参与无核葡萄胚珠败育。研究结果为进一步研究葡萄胚珠败育机制提供基础资料。

英文摘要：

WRKY proteins are a superfamily of plant specific transcription factors and take part in the growth, development and aging process. In our research, the SRAP-cDNA technology was used to scan the genes which have different expression patterns during ovule development between the V. vinifera cv. Thompson Seedless and V. vinifera cv. Pinot Noir. A 311 bp sequence was got which was a part of the predicted sequence of probable WRKY transcription factor 20-like （XM_002272334.2）. then, the full-length sequence （GenBank accession No. JQ782602） was cloned from the ovule of the V. vinifera cv. Thompson Seedless by the RT-PCR, named as VvWRKY20, which was 1 796 bp, with the open reading frame of 1 653 bp, encoding 550 amino acids, containing two WRKY conserved domains, two C2H2 zinc finger domains, two DNA binding sites, and belonged to groupⅠof WRKY family. Expasy-ProtParam analysis showed the molecular weight of VvWRKY20 was 60 140.4 D, the isoelectric point was 6.10, the instability coefficient was 51.01, which proved that VvWRKY20 was an unstable protein. The qRT-PCR was used to analyze the expression of VvWRKY20 in the ovule development of V. vinifera cv. Thompson Seedless and V. vinifera cv. Pinot Noir. The expression levels was relatively stable in V. vinifera cv. Pinot Noir, and the ratio of maximum to minimum was 2.52, while the same ratio was 17.56 in V. vinifera cv. Thompson Seedless. The expression level of VvWRKY20 in V. vinifera cv. Thompson Seedless were higher than that in V. vinifera cv.Pinot Noir at each stages. the ratio of expression in V. vinifera cv. Thompson Seedless to that in V. vinifera cv.Pinot Noir was 15.77 at 10 d, 229.12 at 30 d. These results indicated that VvWRKY20 may be involved in the process of grape ovule abortion.