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在前期通过高通量小RNA测序获得若干在花椰菜器官早期发育过程中呈现差异表达的未知功能miRNA基础上,重点对其中1个在花椰菜下胚轴和子叶中表现为显著差异表达的miRNA(命名为Bra-miR07)进行克隆鉴定及功能分析。研究表明,Bra-miR07前体序列全长为270 bp,可形成稳定的二级发夹结构。Bra-miR07在花椰菜子叶中呈现显著高表达。进一步构建了Bra-miR07的过表达载体并采用浸花法转化拟南芥,获得Bra-miR07过表达的转基因拟南芥株系。表型分析显示,Bra-miR07过表达可显著影响拟南芥植株生长。与野生型相比,过表达Bra-miR07株系表现为生长缓慢,植株矮小,育性降低等发育受阻特征。因此,上述结果预示着Bra-miR07可能也在花椰菜器官发育过程中发挥重要作用。
On the basis of previous studies on several unknown miRNAs that were differentially expressed during early development of cauliflower organs by high-throughput small RNA sequencing, one of the miRNAs that were significantly differentially expressed in cauliflower hypocotyls and cotyledons (named For Bra-miR07) cloning identification and functional analysis. Studies have shown that Bra-miR07 precursor sequence length of 270 bp, can form a stable secondary hairpin structure. Bra-miR07 showed significantly high expression in cauliflower cotyledons. The Bra-miR07 overexpression vector was further constructed and transformed into Arabidopsis thaliana by floral infusion to obtain transgenic Arabidopsis lines with Bra-miR07 overexpression. Phenotypic analysis shows that Bra-miR07 overexpression can significantly affect Arabidopsis plant growth. Compared with the wild-type, Bra-miR07 overexpression showed growth retardation, plant dwarfism and fertility decline. Therefore, the above results indicate that Bra-miR07 may also play an important role in cauliflower organ development.