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为了阐明二化螟Chilo supprressalis滞育幼虫的分子特征及滞育期间保护酶活性的变化规律,本研究应用Trizol法、总量DNA提取法和蛋白定量试剂盒,测定了在长光周期16L∶8D和25℃下发育的非滞育老熟幼虫、在短光周期12L∶12D和25℃下诱导滞育51d的幼虫(称为滞育0个月)、滞育1,2和3个月幼虫的核酸含量和总蛋白含量;同时应用试剂盒测定了老熟幼虫、滞育0,1和2个月的二化螟幼虫5种保护酶(POD,CAT,SOD,LDH和ATP酶)的活性。结果表明:滞育幼虫的总RNA含量显著低于非滞育的老熟幼虫(P<0.05),而滞育1,2和3个月的幼虫之间没有显著差异(P≥0.05)。老熟幼虫的总DNA含量显著高于滞育幼虫(P<0.05)。老熟幼虫的RNA/DNA比值较低,滞育幼虫的RNA/DNA比值较高,RNA/DNA比值随着滞育时间的推移呈现出先上升后下降的趋势。滞育期大于1个月的幼虫中蛋白含量均显著高于非滞育的老熟幼虫(P<0.05),而滞育1,2和3个月的幼虫之间没有显著差异(P≥0.05)。二化螟幼虫体内5种保护酶活性随发育阶段不同而存在差异。滞育幼虫中POD,CAT和SOD的活性随滞育时间延长而逐渐增强,滞育2个月幼虫中的活性最高,而非滞育老熟幼虫中的活性最低;LDH和ATP酶的活性则相反,非滞育老熟幼虫中的活性最高,滞育2个月幼虫中的活性最低。这些结果说明,总RNA和DNA含量降低、RNA/DNA比值先升后降、总蛋白含量升高以及保护酶活性的变化是二化螟幼虫滞育过程中的主要生理特征。
In order to elucidate the molecular characteristics of diapause larvae of Chilo suppressalis and the changes of the protective enzyme activities during diapause, Trizol method, total DNA extraction and protein quantitation kit were used to determine the long-photoperiod 16L: 8D And non-diapause mature larvae developed at 25 ° C, larvae of diapause 51d (referred to as diapause for 0 months) were induced at short photoperiod 12L: 12D and 25 ° C, diapause of 1, 2 and 3 months larvae (POD, CAT, SOD, LDH and ATPase) of larvae of the larvae of the larvae of the larvae of the larvae of the larvae of the larvae of the larvae of the larvae of the rice larvae were measured respectively. . The results showed that the total RNA content of diapause larvae was significantly lower than that of non-diapause mature larvae (P <0.05), while there was no significant difference between diapause larvae of 1, 2 and 3 months (P> 0.05). The total DNA content of mature larvae was significantly higher than that of diapause larvae (P <0.05). The RNA / DNA ratio of mature larvae was lower, and the ratio of RNA / DNA of diapause larva was higher. The ratio of RNA / DNA increased at first and then decreased with the diapause time. The larval protein content of diapause period> 1 month was significantly higher than that of non-diapause mature larva (P <0.05), while there was no significant difference between 1, 2 and 3 month larvae of diapause (P> 0.05 ). The activities of five kinds of protective enzymes in the larvae of S. striiformis vary with the stages of development. The activities of POD, CAT and SOD in diapause larvae gradually increased with the prolongation of diapause period, and reached the highest level in diapause larvae for 2 months, while the activity in larvae of diapause larvae was the lowest. The activity of LDH and ATPase In contrast, larvae of non-diapause were the most active larvae and lagomorphs of 2-month diapause were the lowest. These results indicate that the main physiological characteristics of the larvae of the stem borer, larvae, were the decrease of total RNA and DNA content, the increase of RNA / DNA ratio, the increase of the total protein content and the change of the protective enzyme activity.