欧洲黑蜂和新疆黑蜂越冬期抗氧化酶活性及相关基因表达分析

以欧洲黑蜂和新疆黑蜂为研究对象，比较和分析两个蜂种在越冬条件下抗氧化酶活性和抗氧化能力相关基因表达的变化规律，探究二者在越冬期的抗氧化酶功能及其基因表达特征。本试验测定了蜂群损失率和饲料消耗量等越冬性能指标以及总抗氧化能力（T-AOC）、超氧化物歧化酶（T-SOD）和过氧化氢酶（CAT）等抗氧化酶的酶活力值，并采用实时荧光定量PCR方法检测SOD、CAT、Grx1、MsrA和Vg基因等抗氧化酶功能基因的表达情况。结果显示，欧洲黑蜂和新疆黑蜂的蜂群损失率和饲料消耗率分别为32.86%、32.91%和23.41%、23.91%。在抗氧化酶活性变化规律方面，两个蜂种的T-AOC酶活力呈逐步升高趋势；T-SOD酶活力呈先下降再升高的趋势，在越冬中期达到最低值；CAT酶活力呈先升高再降低又再升高的趋势；在抗氧化功能相关基因的mRNA表达方面，两个蜂种的SOD基因和CAT基因的表达在越冬中期达到最低值；Grx1基因的表达在越冬后期急剧升高，达到峰值；MsrA基因的表达呈先降低后升高的趋势；Vg基因的表达在越冬的中前期呈逐渐升高趋势。结果表明，在低温逆境条件下，两个蜂种体内的抗氧化酶系统可能发挥了重要的防御作用，存在一定的耐寒适应机制，其中越冬中期可能是酶活性变化和基因表达的发生显著变化的特殊阶段。本试验的研究为揭示蜜蜂体内抵御低温应答机制研究提供新线索，也为深入研究蜜蜂耐寒分子机制和抗寒蜂种的选育提供基础数据。

Analysis of antioxidant enzyme activity and gene expression of European Dark bee (Apis mellifera mellifera) and Xinjiang Black bee during wintering period

To explored antioxidant enzymes function and gene expression characteristics of European Dark bee (Apis mellifera mellifera) and Xinjiang Black bee during overwintering period, the indexes of overwintering performances, such as honeybee quantity loss and forage consumption were measured, the activity of antioxidant enzymes T-AOC, T-SOD, CAT and genes expression of SOD, CAT, Grx1, MsrA and Vg were analyzed. The result showed that the honeybee quantity loss and forage consumption of European Dark bee and Xinjiang Black bee were 32.86%, 32.91% and 23.41%, 23.91%. In terms of the rule of antioxidant enzyme activity, the activity of T-AOC in these two species of honeybees increased gradually; the activity of T-SOD showed a trend of descending first and then ascending, reached the lowest value in the middle of overwintering period; the activity of CAT showed a trend of ascending first, then descending and then ascending during the overwintering period. In terms of mRNA expression of genes related to antioxidant function, the gene expression of SOD and CAT of two species reached the lowest value in the middle of overwintering period; The gene expression of Grx1 increased sharply and reached the peak in the late overwintering period; The gene expression of MsrA decreased first and then increased; The gene expression of Vg increased gradually in the middle and early stage of overwintering. These results indicated that the antioxidant enzyme system of two species may play an important defense role in the low temperature stress, there is a certain cold tolerance adaptation mechanism, and the middle overwintering period may be a special stage of significant changes in enzyme activity and gene expression. The research provided new clues for the study on the mechanism of resistant to low temperature in honeybees, and provided basic research data for further research on the molecular mechanism of cold tolerance and the selection for breeding of cold-resistant honeybee species.