Study On Improving The Malnutrition Of Silkworm By Mass Spectrometry Based On Metabolomics

Yunnan has a mild climate all year round and enjoys the reputation of “silkworm breeding paradise”,but with the change of seasons,the quality of mulberry leaves is sharply reduced: in the autumn silkworm period,the incidence of mulberry powdery mildew is over 50%,and the late autumn silkworm period is even higher.100%.The quality of mulberry leaves is reduced,which leads to the survival rate of silkworms falling from 80%-90% to 50%,or even lower.The amount of silkworm cocoons is decreasing,and it is urgent to improve the malnutrition of silkworms.In the previous experiments,FANa was added to the mulberry leaf to raise the silkworm test,FANa to the bacteriostatic test of 9 kinds of bacteria in the diseased silkworm,FANa for the hunger-resistant high temperature survival test of the silkworm,and it was speculated that the brown coal yellow sulphate may be Improve the malnutrition caused by poor quality of mulberry leaves and increase the amount of pupa.On this basis,a model of silkworm malnutrition was established.Whether sodium fulvate enhanced the nutrient absorption of silkworm,the results showed that the addition of FANa compared with the model group reduced the mortality,and the addition of FANa in the mulberry reduction model can be significant.The amount of pupa in the silkworm was increased by 2-3kg(P*<0.05),and the mortality was reduced by 2.5%-5% compared with the control group.It shows that Yunnan brown coal yellow sulphate can enhance the anti-nutrition ability by regulating the metabolism of silkworm.From the overall appearance of the silkworm,it can be confirmed that sodium fulvate can improve the malnutrition of the silkworm.In the mechanism research,firstly,two batches of mulberry leaves plus silkworm feed were collected from June to July and October-November(small powder disease mulberry leaves)in Zhoudong Village,Luliang County,Qujing City,Yunnan Province.The moisture,protein,polysaccharide and fat content of mulberry leaves were measured to find the nutrient content changes of mulberry leaves in different stages of summer silkworm and late autumn.The results showed that there was no significant difference between water content and protein content.In the comparison of fat and polysaccharide content,the mulberry leaf quality in June increased by 28.2% and 82.6%,respectively,compared with the mulberry leaf in November.Further support for sodium fulvate did affect the nutritional metabolism of silkworm.The research on the enhancement of nutrient absorption of silkworm by sodium fulvate was carried out.Firstly,the effect of FANa feeding disease on mulberry leaves was confirmed by silkworm test.The antibacterial test of FANa on nine kinds of bacteria in silkworm was carried out,and FANa was applied to silkworm.Antistarvation and high temperature survival test,and the research mechanism of FANa against the mulberry malnutrition test caused by reducing mulberry.The results showed that the feeding rate of mulberry leaves fed with FANa increased the puerperium rate by 10% compared with the control group;FANa had no significant effect on the bacteria of 9 kinds of silkworm,but it could significantly prolong the survival of silkworm under fasting conditions and high temperature conditions.Compared with the control group,the average survival time of the FANa group increased significantly by 16.72 h and 15.52 h,respectively.In the mulberry-induced malnutrition test,the addition of FANa reduced the mortality rate compared with the model group,and in the mulberry reduction model.Adding FANa can significantly increase the pupa production by 2-3kg(P*<0.05),and the mortality rate is reduced by 2.5%-5% compared with the control group.The above results indicate that Yunnan brown coal yellow sulphate can enhance the anti-nutrition ability by regulating the metabolism of silkworm,and reduce the loss of mulberry powder disease on silkworm breeding.Based on the ideas and techniques of metabolomics,GC-MS was used to study the effect of sodium fulvate on the malnutrition of silkworm,and the relationship between “signature small molecule-biological effect” was constructed,indicating that sodium fulvate was used in the metabolism group of silkworm.Learn the influence,find the key metabolic pathways,and reveal the pharmacological mechanism of fulvic acid on the improvement of silkworm malnutrition.Analysis of differential metabolites in the hemolymph of the normal group(group a),FANa group(group b),mulberry 20%+FANa group(group c)and mulberry 20% group(group d),looking for possible molecular mechanisms The data were processed by GC-MS and XCMS-online.The data were analyzed by PCA and OPLS-DA using SIMPCA.14.The four groups a,b,c and d were completely grouped,and there were significant differences(P< 0.05),indicating that reducing mulberry can affect the basal metabolism of silkworm,sodium fulvate improves the nutritional metabolism pathway of silkworm,and uses S-plot to select metabolites with VIP>1,and obtained four groups a,b,c and d.14 metabolites with significant differences were identified in the NIST14 database: Methyltris(trimethylsiloxy)silane,Dodecamethyl pentasiloxane,Decamethyl cyclopentasiloxane,Octamethyl trisiloxane,Tetradecamethyl hexasiloxane,Dodecamethyl cyclohexasiloxane,Octamethyl cyclotetrasiloxane,3-Acetoxy-3-Hydroxypropionic acid,1-(3,6,6-Trimethyl-1,6,7,7a-tetrah ydrocyclopenta[c]pyran-1-yl)ethenone,Methyl 20-methyl-heneicosanoate Octadecanoic acid,11-met Hyl-hyl ester,Dodecamethyl pentasiloxane,9,12-Octadecadienoic acid(Z,Z)-,methyl ester,Tetradecamethyl cycloheptasiloxane and Decamethyl tetrasiloxane were analyzed by KEGG metabolic pathway through MBRole website,Finally,three metabolic pathways of Biosynthesis of type II polyketide products,Lysine biosynthesis,and Limonene and pinene degradation(limonene and terpene degradation)were obtained.These pathways are closely related to silkworm silkworm silkworm growth and growth.The regulation of pathways and the reduction of mulberry experiments reduced the nutrient composition and the results of the reduction of lipid and polysaccharide content of diseased mulberry leaves.In summary,sodium fulvate has a significant improvement effect on silkworm malnutrition,and its regulation mechanism may be through regulation of secondary metabolic pathway,biosynthesis of type II polyketide products,lysine biosynthesis and limonene and The degradation pathway of pineene affects the synthesis of silk protein and reduces the adverse effects caused by the reduction of polysaccharides and lipid content of mulberry leaves.