航天环境对拟南芥幼苗内参基因表达稳定性的影响

Effects of Space Environment on Stability of Reference Gene Expressions in Arabidopsis Seedling.

目的筛选在航天环境下表达稳定的拟南芥幼苗内参基因。方法"神舟"8号飞船搭载拟南芥幼苗,设置3个试验处理:1)空间飞行处理(F ug);2)空间飞行+1 g离心力处理(F 1g);3)地面对照(G1g)。样品返回后,提取总RNA,通过实时定量RT-PCR的方法,利用geNorm软件分析8个内参基因:ACT8,EF1α,eIF4A,YLS8,UBQ5,ACT2,TIP41和UBC8。结果三个处理中,内参基因表达的稳定性如下:TIP41=UBC>ACT2>UBQ5>YLS8>eIF4A>EF1α>ACT8,其中ACT2,TIP41和UBC的M值小于0.5。结论利用qRT-PCR分析比较太空中拟南芥幼苗基因表达差异时,ACT2,TIP41和UBC可作为内参基因。

Objective To screen the reference gene in Arabidopsis seedling with stable expression under space environment. Methods Shenzhou-8 spacecraft carried Arabidopsis seedlings with three experimental treat- ments ： 1 ） space flight （ F ug） ; 2） space flight ＋ 1 g centrifugal （ F 1 g） ; 3 ） the ground control （ G I g）. Af- ter samples returned, total RNAs were extracted. Real-time quantitative RT-PCR method and geNorm software were used to analyze 8 reference genes：ACTS, EFlα, elF4A, YLS8, UBQS , ACT2, TIP41 and UBC. Re- suits The stability of control gene expression in three treatments was showed as follows： ACTd 〈 EFlα 〈 elF4A 〈 YLS8 〈 UBQ5 〈 ACT2 〈 UBC = TIP41. M-values of ACT2, TIP41 and UBC were less than 0.5. Conclusion With qRT-PCR to analyze and compare the expression differences of Arabidopsis refrence genes in space flight, ACT2, TIP41 and UBC can be used as reference genes.