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MicroRNA(miRNA)在植物对各种逆境胁迫的反应中发挥重要的作用。本研究室在前期研究中,通过基因芯片技术构建了水稻冷胁迫miRNA基因表达谱,并筛选获得了冷胁迫诱导表达的miRNA基因2个——Os-miR1435和Os-miR535。本研究针对Os-miR1435基因,首先采用USERTM技术构建植物超量表达载体,采用农杆菌介导法侵染水稻愈伤组织,获得转基因水稻株系。通过对比野生型和转基因水稻植株的生长发育及农艺性状,发现Os-miR1435基因的超量表达降低了转基因水稻的结实率。通过对比野生型和转基因水稻在种子萌发期和幼苗期冷胁迫处理表型,表明Os-miR1435基因没有影响冷胁迫处理下水稻种子的萌发和幼苗生长。进一步采用psRNATarget软件预测了Os-miR1435的靶基因,通过Real-timePCR分析了靶基因在野生型和转基因植株中的表达量差异,并对靶基因进行了进化分析。本研究将为进一步深入研究Os-miR1435的生物学功能奠定重要的基础。
MicroRNAs (miRNAs) play an important role in the response of plants to various stresses of stress. In our previous study, miRNA gene expression profiles of rice plants were constructed by gene chip technique. Two miRNA genes, Os-miR1435 and Os-miR535, were induced by cold stress. In this study, the Os-miR1435 gene was first used to construct plant overexpression vector using USERTM technology. Agrobacterium-mediated method was used to infect rice callus to obtain transgenic rice lines. By comparing the growth and agronomic traits of wild-type and transgenic rice plants, it was found that over-expression of Os-miR1435 gene reduced the seed setting rate of transgenic rice. By comparing the phenotypes of wild-type and transgenic rice in the germination stage and cold stress in seedling stage, Os-miR1435 gene did not affect the germination and seedling growth of rice under cold stress. Further, psRNATarget software was used to predict the target gene of Os-miR1435. The target gene was analyzed by Real-time PCR in wild-type and transgenic plants, and the target gene was analyzed. This study will lay an important foundation for further study on the biological function of Os-miR1435.