Melcher U, Steffens D L, Lyttle D J, Lebeurier G, Lin H, Choe I S, Essenberg R C
J Gen Virol. 1986 Jul;67 ( Pt 7):1491-8. doi: 10.1099/0022-1317-67-7-1491.
Mutants of cauliflower mosaic virus (CaMV), generated in vitro by modification of recombinant DNA plasmids containing the viral genome, either retained the ability to induce disease symptoms on turnip plants, produced less severe symptoms or failed to induce symptoms. Wild-type symptoms were produced by a variant CaMV DNA of the Cabbage S isolate that had 4 bp in open reading frame (ORF) III replaced with a 16 bp sequence. Less severe symptoms, due to a delay in symptom appearance relative to inoculation with wild-type DNA, were induced by a mutant with a frameshift mutation in ORF II (pSA103). CaMV DNA, recovered from plants infected with pSA103, contained a second mutation which restored the original translation reading frame. Nucleic acid hybridization to 'squishes' of leaf tissue from plants that had been inoculated with mutant DNAs that included DNAs modified in each of the six major ORFs of CaMV DNA revealed that only those plants that appeared diseased had detectable CaMV nucleic acid in uninoculated leaves. Replicated CaMV DNA was also not detected in non-encapsidated and virion DNA fractions from inoculated leaves of non-diseased plants.
通过对含有病毒基因组的重组DNA质粒进行体外修饰而产生的花椰菜花叶病毒(CaMV)突变体,要么保留了在芜菁植株上诱发病害症状的能力,要么产生的症状较轻,要么无法诱发症状。甘蓝S分离株的一种变异CaMV DNA在开放阅读框(ORF)III中有4个碱基对被16个碱基对的序列取代,它产生了野生型症状。由于症状出现相对于接种野生型DNA有所延迟,ORF II中发生移码突变的一个突变体(pSA103)诱发的症状较轻。从感染pSA103的植物中回收的CaMV DNA含有第二个突变,该突变恢复了原来的翻译阅读框。用包括对CaMV DNA的六个主要ORF中的每一个进行修饰的突变DNA接种的植物叶片组织“挤压物”进行核酸杂交,结果表明,只有那些出现病害的植物在未接种的叶片中含有可检测到的CaMV核酸。在未患病植物接种叶片的非衣壳化和病毒粒子DNA组分中也未检测到复制的CaMV DNA。