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为了阐明草地螟Loxostege sticticalis滞育的分子调控基础,本研究应用Trizol法、DNA和蛋白质定量试剂盒、SDS-PAGE电泳技术分别对进入滞育1,2,3和4个月、解除滞育以及非滞育草地螟老熟幼虫中的核酸含量、总蛋白含量和组分的变化进行了测定。结果表明:滞育不同月份幼虫的总RNA含量显著低于非滞育幼虫的总RNA含量(P<0.05);解除滞育幼虫的总RNA含量显著高于滞育2,3和4个月的幼虫,但仍显著低于非滞育的对照组。滞育不同月份、非滞育以及解除滞育幼虫的总DNA含量没有显著差异(P>0.05)。RNA/DNA比值随滞育的开始而显著下降,随着滞育的结束而显著上升。滞育不同月份的幼虫总蛋白含量显著高于非滞育幼虫的总蛋白含量(P<0.05),但解除滞育与非滞育幼虫的总蛋白含量无显著差异。利用SDS-PAGE电泳分析发现滞育幼虫体内存在滞育关联蛋白,蛋白条带在24kDa左右,但非滞育和已经解除滞育的幼虫则没有该蛋白条带。这些结果表明,总RNA含量的降低、RNA/DNA比值下降、总蛋白含量的升高,以及24kDa蛋白的存在是草地螟幼虫滞育的主要生理特征。
In order to elucidate the molecular regulation of diapause of Loxostege sticticalis, Trizol method, DNA and protein quantitation kit and SDS-PAGE electrophoresis were used to detect the diapause of 1, 2, 3 and 4 months of diapause respectively, The nucleic acid content, total protein content and composition of non-diapause grass larvae were determined. The results showed that the content of total RNA in diapause larvae was significantly lower than that in non-diapause larvae (P <0.05). The total RNA content in diapause larvae was significantly higher than that in larvae at 2, 3 and 4 months of diapause Larval, but still significantly lower than non-diapause control group. There was no significant difference in total DNA content between diapause and non-diapause and diapause larvae in different months (P> 0.05). The RNA / DNA ratio decreased significantly with the beginning of diapause and increased significantly with the end of diapause. The total protein content of larvae in different months of diapause was significantly higher than that of non-diapause larva (P <0.05), but there was no significant difference in the total protein content of diapause and non-diapause larvae. SDS-PAGE electrophoresis analysis showed diapause larvae diapause related protein present in the protein band around 24kDa, but non-diapause and diastolic larvae have no such protein bands. These results indicate that the main physiological characteristics of larval larvae of Diapau mask are diapause of the reduction of total RNA content, RNA / DNA ratio, total protein content, and the presence of 24kDa protein.