Jongedijk E, de Schutter A A, Stolte T, van den Elzen P J, Cornelissen B J
MOGEN International nv, Leiden, The Netherlands.
Biotechnology (N Y). 1992 Apr;10(4):422-9. doi: 10.1038/nbt0492-422.
During the last three years we performed field trials to assess levels of resistance against potato virus X (PVX) and changes in intrinsic properties of the potato cultivars Bintje and Escort upon the introduction of the PVX coat protein (CP) gene. Analysis of leaf and tuber samples collected in the field at two week intervals revealed a stable expression of the PVX CP gene throughout the growing season. This resulted in a large decrease in PVX incidence among clonal progeny obtained from previously infected Bintje and Escort clones. Based on evaluation of 50 defined morphological characteristics, tuber yield and grading, 81.8% of the Escort and 17.9% of the Bintje derived transgenic clones proved to be true to type. Overall lightsprout morphology was a useful criterion for the early detection of deviant transgenic clones. Using the polymerase chain reaction (PCR) with convergent primers spanning transgenic sequences, true to type clones could be distinguished unambiguously from the corresponding untransformed cultivars. Clear distinctions between independent transgenic clones could be made by inverted PCR (IPCR) diagnosis revealing integration-specific border fragments. These results demonstrate the commercial feasibility of improving potato cultivars by selectively adding new traits while preserving intrinsic properties, and the possibility of unambiguously identifying independent transgenic cultivars.
在过去三年中,我们进行了田间试验,以评估马铃薯品种宾杰(Bintje)和埃斯科特(Escort)对马铃薯X病毒(PVX)的抗性水平,以及引入PVX外壳蛋白(CP)基因后其内在特性的变化。每隔两周在田间采集叶片和块茎样本进行分析,结果显示PVX CP基因在整个生长季节均稳定表达。这使得从先前感染的宾杰和埃斯科特克隆体获得的克隆后代中PVX发病率大幅降低。基于对50个明确的形态特征、块茎产量和分级的评估,81.8%的埃斯科特和17.9%的宾杰衍生转基因克隆体被证明与原品种一致。总体而言,嫩枝形态是早期检测异常转基因克隆体的一个有用标准。使用跨越转基因序列的收敛引物进行聚合酶链反应(PCR),可以明确区分与相应未转化品种一致的克隆体。通过反向PCR(IPCR)诊断揭示整合特异性边界片段,可以对独立的转基因克隆体进行清晰区分。这些结果证明了在保留内在特性的同时选择性添加新性状来改良马铃薯品种的商业可行性,以及明确鉴定独立转基因品种的可能性。