柑桔黄龙病病原物提纯方法和血清学的初步研究
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中国科学院科学基金


A PRELIMINARY STUDY ON PURIFICATION OF THE PATHOGEN OF CITRUS YELLOW SHOOT AND ITS SEROLOGY
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    摘要:

    从感染柑桔黄龙病的长春花中,用0.1mol/L MgCl_2—0.5mol/L甘露醇-0.6mol/L甘氨酸的混合溶液作黄龙病病原物(YSO yellow sbootO rgancism)分离溶液(pH7.4),然后用纤维素酶酶解法和差速离心相结合的提纯程序,反复试验结果表明该提纯方法重复性稳定,黄龙病病原物(YSO)的提纯产量较高,完整性较好,内外结构清晰。用提纯液备制小鼠腹水抗体,得到微量沉淀反应效价为1:640,环状界凝集反应效性为1:8,免疫电镜效价1:320,琼脂双扩散反应阴性,对照为阴性反应,从而认为YSO具有特异抗原性。

    Abstract:

    A purification method combined with cellulase digestion and differential centri-fugation was found ot purify the pathogen from priwinkle plants infected with citrus yellow shoot,The solution consisting of O.1mol/LMgCl2,O.5 mol/L D-mannitol,O.6mol/L glycine was used as the medium for the isolation and purification.The results obtained in the present study showed that the yield of the pathogen purified was higher,and the external morphology,the transparency af wall-membrane structure and internal structure of the pathogen were better maintained.Meanwhile,the purification method gave consistent results in different replications.The purified pathogen of citrus yellow shoot obtained with the method mentioned abore was used as antigen to immunize mice for the preparation of ascitic fluid.It was shown that the antibodies in the fluid had specific response,The micro-precipitation titer was 1:640,the titer of ring surface agglutination reaction 1:8,and that of immunosorbent electron microscopy 1:320.NO reaction was observed in the agar gel double diffusion assay test.Controls all showed negative reaction.The isolated and purified pathogen was thus recognized to have biological activity of specific antigen.

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柯穗 柯冲.柑桔黄龙病病原物提纯方法和血清学的初步研究[J].华南农业大学学报,1989,(1):1-8

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