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| Cloning and Expression Analysis of the Differentially Expressed VvWRKY20 Gene in the Development of Seedlessness Grape(Vitis vinifera) Ovule Abortion |
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Abstract 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.
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Received: 20 March 2012
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