侵染半夏的两种病毒的分离纯化和初步鉴定

用自然感染的半夏（Ｐｉｎｅｌｌｉａｔｅｒｎａｔａ）为材料，经粗提纯后检查到一种线状病毒和一种球状病毒，用两种方法对担提纯样品中的病毒粒子进行了分离纯化。１０％－７０％连续甘油梯度８００００ｇ离心１５０分钟获得两条病毒带，经紫外吸收测定均为强的核蛋白吸收峰，病毒粒子检查分别为球状和线状病毒粒子，线状病毒经浓缩收集为均一成份．与芋花叶病毒（Ｄａｓｈｅｅｍｍｏｓａｉｃｖｉｒｕｓ，ＤＭＶ）抗体有强的阳性反应。粗提纯样品经０．８％琼脂糖凝胶电脉分离为一条蛋白带，该条带回收后经紫外吸收测定为核蛋白吸收峰，电镜下检查为均一的球状病毒，以ＴＭＶ为对照、醋酸铀（ＵＡＣ）负染后测得该球状病毒（ｐｉｎｅｌｌｉａｓｐｈｅｒｉｃａｌｖｉｒｕｓ１．ＰｓＶ－１）的大小为３１．７ｎｍ；戊二醛固定后磷钨酸（ＰＴＡ）负染测得ＰｓＶ—１的大小为３４．０ｎｍ。各组分经ＳＤＳ－聚丙烯酸胺凝焦电泳分析测得线状病毒和球状病毒的外壳蛋白分子量分别为２０ＫＤ和２８ＫＤ。初步确定线状病毒为ＤＭＶ．球状病毒ＰｓＶ—１为侵梁天南星科半夏的一种新病毒。

Separating and Partial Characterization of Two Viruses from Natural-infected Pinellia Temata

A filament virus and a spherical viruses were detected from partial purifiCation with naretai-infected Pinelliaternata. The preparation was than centrifuged in. 10-70% glycine density-gradient at 80, 000g for 150 minutes, two virus bands, which respectively repressented the spherical viruses and the filament viruses were collected. The filament virus sized 700-800nm long was than identified as dasheen mosaic virus by ISEM test. One coat protein species with a molecular weight of about 20.OKD was determined by SDS-polyacrylamide gel electrophoresis. The spherical virus (pinellia spherical virus 1.PsV-1) was successfully purified by 0.8% agarose electrophoresis. This virus was measured under electron microscope as 31.7nm by UAC stain and 34.0nm by PTA stain. A single coat protein species with a mol. wt. of 28.0KD was detected. PsV-1 must be a new virus infecting Pinellia ternata.