柞蚕微孢子虫体外培养技术体系的建立

Establishment of in vitro Culture System for Nosema pernyi

建立了一套柞蚕微孢子虫株分离及体外增殖保存的技术体系。主要方法是利用柞蚕蛹精巢或卵巢组织制备原代细胞,分离、培养柞蚕微孢子虫;利用柞蚕蛹期滞育可以长期储存的特点,将柞蚕原代细胞分离、培养的柞蚕微孢子虫注射入健康柞蚕蛹体进行继代培养和长期保存;通过原代细胞分离培养、蛹体增殖和保存的交替循环体系,达到分离、增殖及保存微孢子虫株的目的。该技术体系既解决了微孢子虫来源困难,品种混杂不易分离和长期保存等问题,也为进一步探索柞蚕微孢子虫侵染、传播规律和提升柞蚕微粒子病防控效果提供了有力支持。

In this study, a technical system for isolation, proliferation and preservation of Nosema pernyi strains was established.The main method is to isolate and culture N. pernyi by making primary cells from the testis or ovary of Antheraeae pemyi pupa.Using the long-term storage characteristics of A. pemyi pupa at the diapause, the N. pernyi isolated and cultured from the A. pemyi primary cells were injected into healthy A. pemyi pupa for subculture and long-term preservation;the purpose of isolation, multiplication and preservation of Microsporidium strains was achieved through the alternation and circulation system of primary cell isolation, culture and pupa proliferation and preservation.The technical system solved the source of Microsporidium , the difficulty of separation and long-term preservation of hybrid species. It also provided a strong support for further exploring the infection and propagation of N. pernyi and improving the control effect of Microsporidium in A. pemyi .