小麦黄花叶病毒的分离提纯及其血清学等特性

应用梯度离心和超速离心浓缩获得部分提纯的病毒制剂,产量约为7.45g/kg病叶提纯的病毒制剂的紫外吸收曲线呈典型的核蛋白吸收曲线,OD260/OD242和OD260/OD280的比值分别为1.24和1.38。病毒粒子呈线状,宽13—14nm,长度主要分布于250—300nm和550—700nm之间,1000nm以上的粒子也有检到。病毒外壳蛋白仅由一个分子量约为30Kd的亚基组成。在免疫电镜试验中、病毒粒子与日本WYMV抗血清发生强烈的血清学反应。新鲜病叶的超薄切片中可看到大量风轮体和膜状体。

Purification, Serology and Other Properties of Wheat Yellow Mosaic Virus(WYMV)

Wheat yellow mosaic virus(WYMV) has been occurred on wheat in Shanxi province since 1970s. The virus preparation was purified from naturally infected wheat plants by extracting in 0.5mol/L potassium phosphate buffer pH7.0, clarifying in 1/4 v carbon tetrachloride, precipitating with 6% PEG, 3% NaCl and 1% Triton X-100 followed by two cycles of differential centrifugations with 20% sucrose cushions containing 0.3% Triton X-100 and finally centrifuged in 10-50% sucrose density gradients. The yields of virus preparation were about 7.45 mg/kg leaves. Purified virus preparation showed a typical UV spe- ctrum of a nucleoprotetin with max. and min. absorption at 260nm and 242nm, respectively, the ratios of OD_(260)/OD_(242) and OD_(260)/OD_(280) were 1.24 and 1.38. Purified virus preparations contained filamentous particles measuring 13-14nm in width, 250-300nm, 550-700nm in length, and contained a single polypeptide species of mol. wt of c. 30 kd. The virus particles were serologically trapped and decorated by Japanese antiserum against WYMV in immunosorbent electronmicroscopy(ISEM) tests. Numerous cylindrical inclusions(pinwheel) and complex membranous bodies in the cytoplasm of fresh infected wheat leaves were observed with an electron microscope.