Chandra N, Hildebrandt A C
Science. 1966 May 6;152(3723):789-91. doi: 10.1126/science.152.3723.789.
Single tobacco callus cells with or without tobacco mosaic virus inclusion bodies from systemically infected Nicotiana tabacum plants were grown in microcultures. The culture medium consisted of mineral salts and sucrose; it also contained coconut milk. Out of 100 inclusion-bearing cells and 150 inclusion-free cells, 10 and 70 cultured cells divided; eventually 5 and 65 cells, respectively, formed single cell clones. The 5 clones derived from inclusion-bearing cells, and all but 3 of 40 clones from inclusion-free cells, showed virus inclusions in somne cells. The virus could not be detected in three inclusion-free clones by local lesion assay. The results suggest single-cell culture methods for differentiating virus-free plants from cells of pathogen-infected plants.
将来自系统感染烟草植株、带有或不带有烟草花叶病毒包涵体的单个烟草愈伤组织细胞进行微培养。培养基由矿质盐和蔗糖组成；还含有椰乳。在100个含包涵体的细胞和150个不含包涵体的细胞中，分别有10个和70个培养细胞发生分裂；最终分别有5个和65个细胞形成单细胞克隆。来自含包涵体细胞的5个克隆，以及来自不含包涵体细胞的40个克隆中除3个之外的所有克隆，在一些细胞中显示出病毒包涵体。通过局部病斑测定法在3个不含包涵体的克隆中未检测到病毒。这些结果表明了用于从病原体感染植物的细胞中区分出无病毒植物的单细胞培养方法。
