枇杷成花相关基因EjWRKY12序列特性及表达分析

Sequence Characteristics and Expression Analysis of Flowering-related Gene EjWRKY12 in Eriobotrya japonica

WRKY家族是植物中特有且最大的基因家族之一,参与了植物生长发育的各个方面,但在枇杷中尚未见WRKY12的相关报道。研究采用同源克隆法从枇杷芽及叶中克隆得到枇杷EjWRKY12基因,利用生物信息学分析方法对该基因理化性质、蛋白质结构、进化关系、启动子区顺式作用元件等进行分析,并结合转录组数据对其表达模式进行分析。结果表明,枇杷EjWRKY12基因长度为711 bp,共编码236个氨基酸;比对得到的同源基因大多与成花相关,暗示EjWRKY12可能参与枇杷成花;启动子区分析暗示其可能受到光、MeJA、生长素等因素的调控;EjWRKY12蛋白定位在细胞核;外源GA3处理后,叶片中处理组和对照组的EjWRKY12表达量及表达趋势变化不大,但顶芽中对照组的EjWRKY12在成花关键时期表达量最低,在GA3处理后该基因表达量随着处理时间的延长一直上升,暗示其可能是一个抑制成花、促进营养生长的转录因子。本研究克隆得到EjWRKY12基因,对进一步解析枇杷成花调控的分子机制具有指导意义。

The WRKY family is one of the largest and unique gene families in plants,and it is involved in various aspects of plant growth and development,while there weren􀆳t reports on WRKY12 in loquat.In this study,the EjWRKY12 gene was cloned from loquat buds and leaves using homologous cloning,and its physical and chemical properties,protein structure,evolutionary relationship,and cis-acting elements in the promoter region were analyzed using bioinformatics.The expression pattern of the gene was also analyzed in combination with transcriptome data.The results showed that the EjWRKY12 gene in loquat was 711 bp in length and encoded 236 amino acids.The homologous genes obtained by comparison were mostly related to flowering,suggesting that EjWRKY12 may be involved in loquat flowering.Promoter analysis indicated that it might be regulated by light,MeJA,and auxin.EjWRKY12 protein was localized in the nucleus.After exogenous GA3 treatment,there was no significant difference in the expression level and trend of EjWRKY12 in leaves between the treatment group and the control group.However,in the apical bud,the expression level of EjWRKY12 in the control group was the lowest during the critical period of flowering.After GA3 treatment,the expression level of the gene increased continuously with the addition of treatment time,indicating that it may be a transcription factor that inhibits flowering and promotes vegetative growth.This study cloned the EjWRKY12 gene and had guiding significance for further elucidating the molecular mechanism of loquat flowering regulation.