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Cloning and Expression Analysis of CqWRKY1 Gene Induced by Fusaric Acid in chieh-qua[Benincasin hispida(Thunb.)Cogn.var.chieh-qua]

Zhao Qin;Zhang Jinping;Ma Sanmei;He Xiaoming;Xie Dasen

  Based on ESTs of SSH library from interaction analysis between chieh-qua and Fusarium oxysporum f.sp.chieh-qua,full-length cDNA sequence of a WRKY transcription factor(CqWRKY1)was cloned fromA39 FAcultivar treated with fusaric acid by RACE.The GenBank accession number was KX702278.Sequence analysis showed that cDNA sequence length of CqWRKYl was 2 088 bp with one open reading frame of 1 746 bp,encoding 581 amino acids.The deduced amino acids sequence contained two typical conserved domains of WRKY transcription factors and one Zinc finger motif of C_2 H_2(C-X_4-C-X_(22).23-H-X_1-H),which was classified into Class I WRKY transcription factor.Tertiary structure of itsencoding protein included four antiparallel β sheets,three random coils and a zinc-binding pocket at theend of β sheet.CqWRKY1 transcription factor was predicted to be localized in nucleus with two nuclear localization signals of PTKKKVE and PEAKRWR,and possessed transcription,transcription regulation functions.Amino acids sequence alignment and phylogenetic analysis indicated that CqWRKY1 shared high similarities with WRKY transcription factors from other plants,in which the closest evolutionary relationship was shared with WRKYP2 from Cucumis sativus with homology of 92%.The expression patterns of CqWRKY1 in different tissues and treatments were analyzed by real-time quantitative PCR.The results showed that CqWRKY1 expression in chieh-qua exhibited tissue specificity,and the highest level was in roots,followed by stem and leaves.Expression level of CqWRKY1 was strongly induced under fusaric acid stress,signal molecules salicylic acid(SA) and methyl jasmonate(MeJA),suggesting that CqWRKY1 may involved in chieh-qua defense response of fusarium wilt disease,and its regulation may depend on signal pathways mediated by SA or jasmonic acid(JA).……