柞蚕微孢子虫侵染后柞蚕5龄雌雄个体血淋巴蛋白质含量和组成的变化及差异分析

以柞蚕5龄幼虫为材料添食柞蚕微孢子虫(Nosema pernyi,Np),通过测定与分析不同性别5龄幼虫血淋巴蛋白质含量和组成变化的差异,为探究柞蚕对柞蚕微孢子虫感染的免疫应答提供依据。感染柞蚕微孢子虫的不同性别柞蚕5龄幼虫的血淋巴蛋白质含量随蚕体发育进程出现不同的变化:雌性个体与雄性个体分别在添食Np后24 h与48 h,血淋巴蛋白质含量极显著增加(P<0.01),且持续至添食后96 h,其中雌性个体的血淋巴蛋白质含量又极显著高于雄性个体(P<0.01);在添食Np后120 h,雌、雄个体的血淋巴蛋白质含量与对照组无显著性差异(P>0.05)。SDS-PAGE分析表明,添食Np后的柞蚕5龄幼虫血淋巴蛋白质组成与对照组基本一致,均显示出20条分子质量在20.1～97.2 kD的蛋白条带,其中雌、雄个体在添食后96 h,大小约44 kD的蛋白条带明显加深,大小约28 kD的蛋白条带雌性个体在添食后144 h、雄性个体在添食后120 h明显加深,且雄性个体在添食后120～168 h大小约42 kD的蛋白条带明显加深,但在添食后192 h与对照组无明显差异。研究结果说明Np侵染后,柞蚕5龄幼虫血淋巴蛋白质的含量及组成均产生了一定的变化,并且雌雄个体间的变化存在差异。

Variation and Difference Analysis on Content and Composition of Proteins in Haemolymph of the Fifth Instar Antheraea pernyi Larvae Infected by Nosema pernyi

In order to provide evidence for probing into immune response of Antheraea pernyi to Nosema pernyi(Np) infection,changes of the content and composition of haemolymph proteins in the 5th instar female and male Antheraea pernyi larvae that were administrated with Np spores were measured and analyzed.The results showed that contents of hemolymph proteins in the 5th instar female and male Antheraea pernyi larvae infected with Np spores varied differently with larval development.The hemolymph protein contents in female and male larvae increased extremely significantly(P<0.01) at 24 h and 48 h after Np administration respectively,and lasted till 96 h.What’s more,the content of haemolymph proteins in female larvae was extremely significantly higher than that of the male(P<0.01).At 120 h after infection,the haemolymph protein contents in both female and male larvae had no significant difference with the control group(P>0.05).SDS-PAGE analysis revealed that the composition of haemolymph proteins in the 5th instar larvae infected by Np spores was similar to that of the control group,both showing 20 protein bands with molecular weights ranging from 20.1 to 97.2 kD.The protein bands of about 44 kD in haemolymph of the female and male larvae were obviously thicker at 96 h after Np infection.The protein band of about 28 kD was obviously thicker at 144 h in female larvae and at 120 h in male larvae after Np infection.Moreover,the protein band of about 42 kD in male larvae was obviously thicker at 120~168 h after Np infection,but showed no obvious difference with the control group at 192 h.These experimental results suggest that there are certain changes in contents and compositions of haemolymph proteins in the 5th instar Antheraea pernyi larvae,and the changes in female larvae are different with those in male ones.