青花菜转录因子基因BoiWRKY8及其与抗病性的关系OA北大核心CSTPCD

WRKY transcription factors are a distinct class of regulatory proteins found exclusively in plants,and they play an important role in combating various biotic and abiotic stresses.In this study,based on the isolation of BoiWRKY8 gene from broccoli,bioinformatic methods for sequence analysis were utilized,and qRT-PCR was applied to confirm the expression pattern of this gene under the infection of Sclerotinia sclerotiorum and Botrytis cinerea.Furthermore,overexpressed plants were subjected to disease resistance identification,aiming to preliminarily elucidate the function of BoiWRKY8 in disease responses.The results indicated that the genomic sequence of BoiWRKY8 was 3 170 bp in length,containing two introns of 776 bp and 1 422 bp.The complete coding sequence was 972 bp,encoding 323 amino acids.The WRKY domain of BoiWRKY8 was composed of 60 amino acid residues,including a WRKYGQK sequence followed by a C2H2 zinc finger structure of C-X4-C-X23-H-X1-H.Phylogenetic analysis results revealed that BoiWRKY8 clustered together with its homologous sequences from Brassica genus with a support value of 100％,and exhibited the closest relationship with wild cabbage.qRT-PCR results showed that the expression of BoiWRKY8 was significantly induced by the necrotrophic fungal pathogens S.sclerotiorum and B.cinerea,and the highest transcript levels were observed at 6 h and 12 h after inoculation,with fold changes of 3.18/2.68 and 3.22/2.90,respectively.BoiWRKY8 overexpressing broccoli plants exhibited increased resistance to both S.sclerotiorum and B.cinerea,concomitant with elevated expression of BoPDF1.2,one marker gene of the JA/ET pathway.Isolation and expression analysis of BoiWRKY8 gene provides a foundational framework for further investigation into the underlying mechanisms of broccoli's resistance to diseases and molecular breeding.