采用荧光定量PCR(Real-time PCR),进行miR167及其靶基因ARF8表达水平的相对定量分析,初步得出水稻愈伤组织发育过程中Auxin-miR167-ARF8信号通路的调控情况.发现经过12h~24h的积累,2mg/L的2,4-D能够明显地上调miR167的表达水平,从而导致miR167的靶基因ARF8的表达量下降,这种变化在第7天的时候趋于平缓,直到形成愈伤组织,miR167和ARF8的表达量维持在一个相对稳定的水平.本实验为Auxin-miR167-ARF8信号通路的研究提供了切入点.
Though relative quantitative analysis on the expression level of miR167 and its target gene ARF8 by SYBR GREEN Real-time PCR, the preliminary results about the regulation and control situation of Auxin-miR167-ARF8 signal pathway were got during the growth and development process of rice callus. It's found that after 12 - 24 hours, the expression level of miR167 was up-regulated obviously, resulting in the decline of expression quantity of ARF8 which was the target gene of miR167. This changing tendency was flattened after the seventh day. The expression level of miR167 and ARF8 maintained at a relative stable level until the formation of callus. This experiment provided an enter-point of the further study on Auxin -miR167 -ARF8 signal pathway.