Differences Of Metal Complexes Between The Blood Of Resistance And Non-resistant Silkworm Under BmNPV Stress

Silkworm mulberry is an important economic industry and has more than five thousand years of history in China.BmNPV(Bombyx mori nuclear polyhedrosis virus)is the main disease virus which causes a variety of silkworm disease and great economic loss in sericulture.Scientists have screened and cultivated a variety of resistant silkworm varieties and put into the sericulture production.At present,there are many studies on the infection and resistance of BmNPV.The main research directions are silkworm genomics and proteomics.Some scholars have applied transgenic technology to the resistance of BmNPV.However,it has not been reported that the mechanism of silkworm resistance to BmNPV from metalloproteins.Metalloproteomics is an important part of Proteomics,which focuses on the function and their distribution in organisms of metal ions bounding to proteins.Studing of metalloproteomics often require specific isolation technique to complete qualitative and quantitative analysis of metalloproteins.In this study,silkworms with and without BmNPV resistance were chosen as experiment materials and their fifth-instar larvae were inoculated with BmNPV.Inductively coupled plasma mass spectrometry(ICP-MS)was applied for the detection of 9 kinds of metallic elements in the heamolymph of silkworms,then size exclusion-high performance liquid chromatography(SEC-HPLC)was coupled with ICP-MS for the separation and determination of the metalloproteins in silkworms.It provides a more simple and effective method for the separation and analysis of metal protein in silkworm.The results were as follows:1.The content of Cu?Mn?Zn and Rb in silkworm were the same as those of resistant and non-resistant varieties.The contents of K?Ca?Mg?Fe and Ni in the blood of non-resistant silkworms were significantly different from the second day after feeding BmNPV.K?Ca?Mg?Fe and Ni were significant difference on the fifth day after feeding BmNPV.2.The SEC results showed that the spectrum of the resistant and non-resistant control groups was significantly different at 8 min on the third day after feeding the virus.Compared with the resistance silkworm control group,one of the peaks wasundetectable after BmNPV infection.On the fifth day of the fifth instar,Compared with the control group,the 8min peak of BmNPV was significantly changed.two peaks of 10-12 min were not detected after BmNPV infection.3.The SEC-ICP-MS analysis of four groups of silkworm hemolymph was compared and analyzed.It was found that the 11 min peak of non-resistant silkworm was almost undetectable after infection with non-resistant silkworm control group.The SEC-ICP-MS patterns of Mn ? Zn and Cu in the blood of four groups were significantly different on the fifth day after feeding the virus.4.the 8min fraction was separated and collected by the dextran G100 gel column.it was proved that the separation method for the separation and collection of the 8 min fraction using the semi-prepared dextran gel column was higher than that of the conventional HPLC-fraction collector,and it was not necessary to re-concentrate the fractions to provide the study of silkworm metal protein more convenient and effective separation of the collection method.