家蚕微孢子虫孢子的异质性研究：基于单细胞拉曼光谱的分析  

Investigation on the Heterogeneity of Nosem a bombycis Spores: Insights from Single-cell Raman Spectroscopy

作　　者：王桂文[1] 余裕超 黄旭华 黄深惠 苏丽[2] 

机构地区：[1]广西科学院,南宁530007 [2]广西大学农学院,南宁530004 [3]广西壮族自治区蚕业科学研究院,南宁530007

出　　处：《蚕业科学》2016年第5期856-864,共9页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31460035,11264004,31560048)

摘　　要：家蚕微粒子病是由家蚕微孢子虫(Nosema bombycis,Nb)感染、寄生而引起的对蚕业生产具有毁灭性的疫病,深入解析Nb孢子的基础生物学特性和异质性对病害的有效防控具有重要意义。通过拉曼光谱技术采集不同感病家蚕个体内的单个Nb孢子的拉曼光谱信息,应用化学计量学分析Nb孢子光谱的同质性和异质性。结果发现,来自同一感病家蚕个体内的Nb样本可观察到几种类型的拉曼光谱,成熟Nb孢子中最为显著的拉曼峰主要来自孢内的海藻糖;通过主成分分析,来自不同感病家蚕个体的Nb孢子间的拉曼光谱差异主要体现在海藻糖峰和脂类物质峰,而且在部分感病家蚕个体来源的Nb孢子之间,其拉曼光谱中的海藻糖和核酸、蛋白质的峰强度甚至具有显著差异。研究结果表明,无论是来自同一感病家蚕个体的Nb孢子间,还是来自不同感病家蚕个体的Nb孢子间,都有一定的异质性,利用拉曼光谱可以在单细胞水平上便捷地分析微孢子虫孢子的基础生物学特性和异质性。Pebrine is an infectious disease of silkworm（ Bombyx mori） caused by Nosema bombycis（ Nb）,bringing heavy loss to sericulture once occurred. Deeper understanding of the biological characteristics and heterogeneity of Nb spores is important for prevention and control of pebrine disease. In present study,Raman spectroscopy was used to record the Raman spectra of individual Nb spores isolated from diseased silkworm and chemometrics was employed to analyze the homogeneity and heterogeneity of spore spectra. Several types of Raman spectral curves were found among spores isolated from the same diseased silkworm body. The most significant Raman peaks of mature spore were mainly generated by trehalose in the spore. Principal component analysis showed that the spectral difference of spores isolated from different diseased silkworm bodies was basically displayed in Raman peaks generated by trehalose and lipids. Moreover,contents of trehalose,nucleic acids and proteins differed significantly in spores isolated from different diseased silkworm bodies. These results show that heterogeneity exists among Nb spores isolated from the same diseased silkworm body and from different diseased silkworm bodies,and Raman spectroscopy can be used to quickly characterize the basic biological properties and heterogeneity of microsporidian spores at the single cell level.

关 键 词：拉曼光谱 家蚕微孢子虫 单细胞 异质性 

分 类 号：S884[农业科学—特种经济动物饲养]