三个甘蓝 WRKY 基因的克隆及其对非生物胁迫的表达.

WRKY proteins are a class of transcription factors involved in plant growth and development, biotic and abiotic stress, and other biological processes. To study the response mechanism of cabbage WRKY genes under stress of adversity may lay the foundation for further research on the stress tolerance function of BoWRKYs. BoWRKY40 (BolC02g035760.2Jl, BoWRKY46 (BolC04g031460.2Jl,and BoWRKY70 (BolC04g035730.2Jlfrom cabbage (Brassica oleracea var. capitata L.lseedlings were cloned by PCR approach. The pSuper1300: BoWRKYs-GFP expression vector was constructed by homologous recombination method for subcellular localization. Response patterns of BoWRKYs genes to abscisic acid (ABAl, PEG8000 and salt stress were analyzed by RT-qPCR. As results, the length of CDS regions of BoWRKY40, BoWRKY46 and BoWRKY70 were 873, 831 and 864 bp, which encoded proteins of 290, 276 and 287 amino acids,respectively. The predicted molecular weights of BoWRKY proteins were 32.41, 32.08 and 32.52 kD with theoretical isoelectric points (pIlof 6.77, 6.18 and 5.62. The conserved structural domain of the WRKY superfamily ‘WRKYGQK’ was present in the proteins, and multiple alignment analysis revealed that BoWRKYs had high conservation with WRKYs in Arabidopsis (Arabidopsis thalianal, tomato (Solanum lycopersicum L.l, maize (Zea mays L.), and cotton (Gossypium hirsutum L.). Subcellular localization analysis showed that BoWRKYs were located in the nucleus. Fluorescence quantitative PCR results showed that WRKY family genes were induced in different degrees under ABA,PEG8000 and salt stress. Among them, BoWRKY40 responded positively to three kinds of stress, BoWRKY46 and BoWRKY70 were up-regulated under ABA and salt stress, and their expressions gradually decreased after PEG8000 treatment. These results indicated that the expressions of cabbage BoWRKY40, BoWRKY46, and BoWRKY70 were closely related to stress responses. [ABSTRACT FROM AUTHOR]