黄粉虫抗菌肽的诱导·分离及其抗菌活性研究

Study on Induction,Isolation and Antibacterial Activity of Antibacterial Peptides from Tenebrio molitor

[目的]探讨不同方法诱导对黄粉虫抗菌肽活性的影响,为黄粉虫抗菌肽的修饰改造、分子克隆、外源表达等奠定基础。[方法]选7龄、8龄、9龄的黄粉虫幼虫,以巴氏杆菌和沙门氏菌为诱导菌采用微量注射法、针刺法诱导抗菌肽,对不同菌液浓度和不同方法诱导的不同龄期黄粉虫幼虫抗菌肽的抑菌效果进行研究。[结果]①黄粉虫龄期对抗菌肽活性有显著影响。采用巴氏杆菌和沙门氏菌作为诱导菌,对7龄、8龄和9龄黄粉虫进行诱导,在诱导处理中,8龄幼虫诱导的抗菌肽活性显著大于其他龄期幼虫诱导的抗菌肽活性(P<0.05)。②不同菌液浓度诱导黄粉虫幼虫产生的抗菌肽抑菌活性存在差异。菌液浓度为对数期浓度稀释100倍时抗菌肽活性显著大于其他稀释倍数诱导的抗菌肽活性(P<0.05)。③不同方法诱导产生的抗菌肽抑菌活性存在差异。微量注射诱导的抗菌肽活性显著大于针刺诱导的抗菌肽活性(P<0.05)。④2种细菌诱导的抗菌肽活性有显著差异。采用沙门氏菌诱导的抗菌肽最高活性显著大于采用巴氏杆菌诱导的抗菌最高活性(P<0.05)。[结论]黄粉虫抗菌肽的活性受黄粉虫龄期、诱导源微生物种类、菌液浓度、诱导方法的影响。该研究中,以8龄黄粉虫为试验动物,采用对数期沙门氏菌液稀释100倍进行微量注射法诱导,所获得的抗菌肽活性最好。

[Objective]The purpose of this study was to explore the effects of different induction methods on the activity of Tenebrio molitor antimicrobial peptides,and lay the foundation for modification,molecular cloning,and exogenous expression of Tenebrio molitor antimicrobial peptides.[Method]The 7,8 and 9 instar larvae of Tenebrio molitor were selected for this study.Antibacterial peptides were induced by Pasteurella and Salmonella.Microinjection and acupuncture methods were used to study the antibacterial effects of antibacterial peptides induced by different concentrations of bacteria and different methods in Tenebrio molitor larvae of different ages.[Result]The results showed that:①The age of Tenebrio molitor had significant effect on the activity of antimicrobial peptides.The activity of antibacterial peptides induced by 8th instar larvae was significantly higher than that induced by other instar larvae(P<0.05).②The antibacterial activity of antibacterial peptides induced by different concentrations of bacterial solution was different.When the concentration of bacterial solution was diluted 100 times of the logarithmic phase concentration,the antibacterial peptide activity was significantly higher than that induced by other dilution multiples(P<0.05).③The antibacterial activities of antibacterial peptides obtained by different induction methods were different.The activity of antibacterial peptides induced by microinjection was significantly higher than that induced by acupuncture(P<0.05).④The activity of antibacterial peptides induced by different bacteria was significantly different.The highest activity of antimicrobial peptides induced by Salmonella was significantly higher than that induced by Pasteurella(P<0.05).[Conclusion]The activity of antimicrobial peptide of Tenebrio molitor is affected by the age of Tenebrio molitor,the type of microorganism,the concentration of bacterial solution and the method of induction.In this study,Tenebrio molitor at the age of 8 was used as the experimental animal,and was induced by microinjection of salmonella solution diluted 100 times in logarithmic phase,and the antibacterial peptide activity obtained was the best.