岩黄连DELLA基因编码区的克隆及蛋白转录自激活能力检测OA

Corydalis saxicola,a perennial herbaceous plant of the Papaveraceae family in the genus Corydalis,grows exclusively in karst rock crevices.C.saxicola is also a Chinese national second-class protected plant.DELLA proteins,plant-specific transcriptional factors,play crucial regulatory roles in plant growth and development.In order to explore the function of DELLA proteins in the growth and development of C.saxicola,coding sequence(CDS)of two DELLA-like genes,CsDELLA1 and CsDELLA2,were identified and amplified based on full-length transcriptome and transcriptome data of different tissues of C.saxicola.Molecularly,the physical and chemical properties,structures,the relative expression levels of their transcripts,and key domain transcriptional autoactivation of the two DELLAs were analyzed.The results were as follows:(1)The length of CsDELLA1 and CsDELLA2 CDS were 1 902 bp and 1 614 bp,encoding 633 amino acids(aa)and 537 aa,respectively.(2)Quantitative real-time PCR results showed that the relative expression levels of the two DELLA transcripts were exclusively high in the leaves of C.saxicola.(3)Amino acid sequence homology alignment and phylogenetic tree analysis revealed that both CsDELLA1 and CsDELLA2 possessed conserved DELLA domain and GRAS domain,and both had high similarities with members of the DELLA family of Papaveraceae.(4)The analysis results of protein structures revealed that α-helices were important components of the protein structures of CsDELLA1 and CsDELLA2,and their three-dimensional spatial arrangements were similar,too.(5)Both CsDELLAs exhibited transcriptional autoactivation capacities,but the activation domain sites differed.Specifically,CsDELLA1,CsDELLA2,as well as truncated proteins CsDELLA1-N52-119 aa,CsDELLA1-N120-259 aa,CsDELLA1-C260-633 aa,and CsDELLA2-C93-537 aa,had transcriptional autoactivation capacities;while N-terminal sequence of the two CsDELLA1-N1-51 aa and CsDELLA2-N1-92 aa showed no autoactivation capacity.This study lays a foundation for further exploration of the interactions between CsDELLAs and other transcription factors,as well as their biological functions in the growth and development of C.saxicola.