摘要
miRNA(microRNA)是一类长度为18~25nt的内源性非编码小分子RNA,通过对其靶基因mRNA的降解或抑制翻译来调控基因表达,进而参与调控植物相关的生理活动。该研究分析gma-miR1507a的成熟体序列、茎环结构和前体启动子区域的顺式作用元件并检测了逆境处理下大豆组织中gma-miR1507a的水平;使用psRNATarget在线软件预测了gma-miR1507a的靶基因;构建植物表达载体amiRNA1507a-pCAMBIA2301并转化大豆子叶,获得了发状根。结果发现,gma-miR1507a前体的启动子区域存在与干旱胁迫和病菌侵染相关的顺式作用元件;在线软件psRNATarget预测到9个gma-miR1507a的靶基因;该研究利用发状根介导的遗传转化结合GFP染色,确定阳性发状根;在转基因根毛中,gma-miR1507a水平与空载相比显著提高,预测的部分靶基因表达量显著下调。以上结果显示,过表达amiRNA1507a能够提高大豆发状根中gma-miR1507a水平,为研究gma-miR1507a的功能奠定基础。同时,gma-miR1507a可能参与调控大豆的逆境胁迫。
miRNA（microRNA） are a class of endogenous and noncoding small molecular RNA with approximately 18-25 nucleotides（nt） in length. They can regulate gene expression through mRNA cleavage or translation inhibition. We analyzed the mature sequence, stem-loop structure and promoter sequence of gmamiR1507 a. The level of gma-miR1507 a in soybean tissue under stress was tested by stem-loop RT-PCR method. The target genes of gma-miR1507 a were predicted by the online software ps RNATarget. Then, we constructed artificial miRNA expression vectors-miRNA1507a-p CAMBIA3301. Furthermore, amiRNA1507 a gene was overexpressed in soybean cotyledon and the transgenic hairy roots were obtained. The results showed that there were a number of cis-acting elements related to drought stress and pathogen infection in the promoter sequence of precursor gma-miR1507 a and 9 target genes of gma-miR1507 a were predicted. We obtained the transgenic hairy roots by hairy root-mediated genetic transformation system and GFP staining. The expression level of gmamiR1507 a in the transgenic hairy roots significantly increased compared with that in empty vector. The expression of seven predicted target genes significantly reduced in transgenic hairy roots. Based on the results, the transgenic hairy roots of amiRNA1507 a overexpression could increase the expression of gma-miR1507 a which could be used to analyze the function of gma-miR1507 a. In addition, gma-miR1507 a may involve in biotic stresses responses in soybean.
出处
《中国细胞生物学学报》
CAS
CSCD
2017年第7期906-915,共10页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:31301340)
江苏省自然科学基金(批准号:BK20130684)
国家公益性行业(农业)科研专项经费(批准号:201303018)
转基因生物新品种培育重大专项经费(批准号:2016ZX08004002-005)
长江学者和创新团队发展计划(批准号:PCSIRT13073)
大豆现代产业技术体系(批准号:CARS-004-PS10)
江苏省现代作物生产协同创新中心资助的课题~~
