苋菜试管开花过程中相关microRNA表达的qPCR分析

以苋菜无菌试管苗及其试管开花过程中各阶段培养物为材料,利用qPCR技术进行苋菜试管开花过程相关microRNA的表达分析。研究结果表明:由NormFinder软件计算出最适合的内参基因为ama-miR159a;在苋菜试管开花过程中,ama-miR156c、ama-miR156d、ama-miR156g相对表达量在幼苗期向壮苗期转变过程中呈下降趋势,并在整个发育过程中一直保持较低水平;ama-miR156h、ama-miR157b、ama-miR157c、ama-miR157d、ama-miR3和ama-miR10相对表达量在幼苗期向花芽期转变过程中呈下降趋势,并在花芽期达到最低;ama-miR172d的表达模式与miR156/miR157表达模式相反;苋菜试管苗幼苗期到花芽期形成时ama-miR319b相对表达量呈上升趋势,ama-miR164b在花芽期时相对表达量比较高,花芽期向开花期转变时,其相对表达量降低;ama-miR166a/miR166g表达在苋菜开花过程中始终呈上调表达;ama-miR167b/miR167d和ama-miR4相对表达量先略有升高然后降低,在花芽期降到最低,随后又升高。由此可见,microRNA在调控苋菜试管苗由幼苗期向壮苗期转变、营养生长向生殖生长转变以及花器官的形成等过程中起重要的调控作用。

Expression Analysis of Related microRNAs During the in vitro Flowering by qPCR in Amaranthus mangostanus L.

In this experiment in vitro plantlets and the cultures during the in vitro flowering were used as the material to study the expression of related microRNAs during the in vitro flowering by qPCR in Amaranthus mangostanus L. The results showed that the most suitable reference gene was ama-miR159a by NormFinder software analysis. The relative expression levels of ama-miR156c, ama-miR156d and ama-miR156g were downtrend from the young seedling stage to the adult seedling stage, and constantly kept at lower levels during the whole developing process. The relative expression levels of ama-miR156h, ama-miR157b, ama-miR157c, ama-miR157d, ama-miR3 and ama-miR10 were downtrend from the young seedling stage to the flower-bud stage, and decreased to the lowest level at the flower-bud stage. The ama-miR172d showed the opposite expression model with miR156/miR157. The relative expression level of ama-miR319b was upstrend from the young seedling stage to the flower-bud stage. The relative expression level of ama-miR164b was higher at flower-bud stage, then decreased. The relative expression levels of ama-miR166a/miR166g were up-regulation during the whole flowering period. The relative expression levels of ama-miR167b/miR167d and ama-miR4 slightly increased at first and then decreased,and to the lowest at the flower-bud stage, finally increased. Therefore, the microRNAs played the important role in regulating the conversion from the young seedling stage to the adult seedling stage, from the vegetative growth to the reproductive growth and formation of floral organs and so on.