Cloning,expression,and bioinformatics analysis of KoWRKY43 gene in Kandelia obovata
The transcription factor WRKY is widespread in flowering plants,and regulates plant growth,development and defense responses.To explore the role of the WRKY gene in Kandelia obovata under abiotic stresses,total RNA was extracted from the leaves of K.obovata,and then the KoWRKY43 gene(GenBank accession number OR789874)was cloned by reverse transcription-PCR(RT-PCR);the gene sequence and protein structure were analyzed by bioinformatics,and its expression pattern was studied by real-time fluorogenic quantitative PCR(qRT-PCR).The results showed that the gene contained an open reading frame(ORF)of 942 bp,encoding 313 amino acids.The pro-tein KoWRKY43,with a molecular formula of C1484H2415N439O459S14,a molecular weight of 34.2 ku,and a theoreti-cal isoelectric point of 9.74,had no signal peptide or transmembrane structure,and it was predicted to be located in the nucleus.Phylogenetic analysis indicated that KoWRKY43 had the most homology with Manihot esculenta,Popu-lus alba and Salix suchowensis.The qRT-PCR analysis showed that the expression abundance of KoWRKY43 gene was highest in roots,which was significantly higher than that in stems,leaves,flowers and fruits.The expression level of the KoWRKY43 in young leaves was induced by NaCl,salicylic acid(SA)and abscisic acid(ABA),and the high-est expression levels were observed at 24 h,6 h and 6 h,respectively.However,methyl jasmonate(MeJA)did not significantly alter the expression level of KoWRKY43 within 24 h.The present research provided a theoretical basis for subsequent functional studies of the gene and the cultivation of stress-resistant K.obovata varieties.
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