Vetten H J, Lesemann D E, Maiss E
Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Biochemie und Pflanzenvirologie, Braunschweig, Federal Republic of Germany.
Arch Virol Suppl. 1992;5:415-31. doi: 10.1007/978-3-7091-6920-9_44.
The serological relationships among strains of bean common mosaic virus (BCMV) (genus Potyvirus, family Potyviridae) were investigated by testing 13 isolates of the 10 known BCMV pathotypes with two monoclonal antibodies and six antisera to BCMV strains. In addition, other properties of serologically distinct BCMV strains were compared. Two groups of BCMV strains were obtained by ELISA and Western blot serology: serotype A contained the BCMV strains NL3, NL5, and NL8 and serotype B contained the BCMV strains NL1, NL2, NL4, NL6, US4, NL7, NY15, and Fla. SDS polyacrylamide gel electrophoresis and Western blotting of freshly purified preparations, and of extracts from leaves infected with eleven BCMV strains showed that the apparent molecular mass of the capsid protein of the serotype A isolates NL3, NL5, and NL8 are lower (about M(r) 33,000) than those of the serotype B isolates (M(r) 34,500 to 35,000). The normal lengths of the particles of the serotype A isolates were shorter (810-818 nm) than those of most isolates (except NL6 and NY15) of serotype B (847-886 nm). All isolates studied induced cytoplasmic pinwheel and scroll inclusions. Cells infected with serotype A isolates contained a specific type of proliferated endoplasmic reticulum which was never found in cells infected with serotype B isolates. The capsid protein gene of a representative member of each serotype was cloned and sequenced. Molecular mass calculations based upon nucleotide sequence-derived amino acid sequences yielded M(r) of 29,662 and 32,489 for the capsid proteins of the serotype A isolate NL8 and the serotype B isolate NL4, respectively. Comparison of the coat-protein sequences showed considerable differences at the N-termini whereas the core regions and the C-termini appeared to be highly conserved. Marked differences were also observed within the 3' non-coding regions of cloned cDNAs of NL 4 and NL 8. The striking differences between the two serotypes of BCMV strongly suggest that they be classified as two distinct potyviruses which naturally infect Phaseolus beans.
通过使用两种单克隆抗体和六种针对菜豆普通花叶病毒(BCMV,马铃薯Y病毒属,马铃薯Y病毒科)菌株的抗血清,对10种已知BCMV致病型的13个分离株进行检测,研究了BCMV菌株之间的血清学关系。此外,还比较了血清学上不同的BCMV菌株的其他特性。通过酶联免疫吸附测定(ELISA)和蛋白质免疫印迹血清学获得了两组BCMV菌株:血清型A包含BCMV菌株NL3、NL5和NL8,血清型B包含BCMV菌株NL1、NL2、NL4、NL6、US4、NL7、NY15和Fla。对新鲜纯化制剂以及感染了11种BCMV菌株的叶片提取物进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和蛋白质免疫印迹分析,结果表明,血清型A分离株NL3、NL5和NL8的衣壳蛋白表观分子量较低(约为M(r) 33,000),低于血清型B分离株(M(r) 34,500至35,000)。血清型A分离株的病毒粒子正常长度较短(810 - 818 nm),比血清型B的大多数分离株(除NL6和NY15外)(847 - 886 nm)短。所有研究的分离株都诱导产生细胞质风轮状和卷丝状内含体。感染血清型A分离株的细胞含有一种特定类型的增殖内质网,而在感染血清型B分离株的细胞中从未发现过这种内质网。对每种血清型的一个代表性成员的衣壳蛋白基因进行了克隆和测序。根据核苷酸序列推导的氨基酸序列进行的分子量计算得出,血清型A分离株NL8和血清型B分离株NL4的衣壳蛋白分子量分别为M(r) 29,662和32,489。衣壳蛋白序列的比较表明,N端存在相当大的差异,而核心区域和C端似乎高度保守。在NL 4和NL 8的克隆cDNA的3'非编码区域也观察到明显差异。BCMV两种血清型之间的显著差异强烈表明,它们应被归类为两种不同的马铃薯Y病毒,它们自然感染菜豆属豆类。