Biologisches Institut, Abteilung für Molekularbiologie und Virologie der Pflanzen, Universität Stuttgart, Pfaffenwaldring 57, D-70550 Stuttgart, Germany.
Virus Res. 2011 Jun;158(1-2):169-78. doi: 10.1016/j.virusres.2011.03.029. Epub 2011 Apr 5.
Beet curly top virus (BCTV) limits sugarbeet production considerably. Previous studies have shown that infections are associated with the generation of defective DNAs (D-DNA) which may attenuate symptoms. Transgenic sugarbeet lines were established carrying a partial direct repeat construct of D-DNA in order to examine whether they are useful as a means of generating tolerance against BCTV. Thirty four independent transgenic lines were challenged. Viral full-length and D-DNAs were monitored by polymerase chain reaction (PCR) or rolling circle amplification (RCA) and restriction fragment length polymorphism (RFLP). The differential accumulation of both DNA species was compared with symptom severity during the course of infection. RCA/RFLP allowed the discrimination of two D-DNA classes which were either derived from the transgenic construct (D(0)) or had been generated de novo (D(n)). The statistical analysis of the results showed that the presence of D(0)-DNA correlated with increased symptom severity, whereas D(n)-DNAs correlated with attenuated symptoms.
甜菜曲顶病毒（BCTV）极大地限制了甜菜的产量。先前的研究表明，感染与缺陷 DNA（D-DNA）的产生有关，D-DNA 可能会减轻症状。为了研究携带 D-DNA 部分直接重复构建体的转基因甜菜品系是否可作为产生对 BCTV 耐受性的手段，建立了转基因甜菜品系。对 34 个独立的转基因系进行了挑战。通过聚合酶链反应（PCR）或滚环扩增（RCA）和限制性片段长度多态性（RFLP）监测病毒全长和 D-DNA。在感染过程中，比较了两种 DNA 物质的差异积累与症状严重程度的关系。RCA/RFLP 允许区分源自转基因构建体的两种 D-DNA 类（D(0)）或从头产生的 D-DNA 类（D(n)）。结果的统计分析表明，D(0)-DNA 的存在与症状严重程度的增加相关，而 D(n)-DNA 与症状减轻相关。
