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首页> 外文期刊>Horticulture,Environment,and Biotechnology >Transformation of Solanum tuberosum L. with potato leafroll virus (PLRV) coat protein gene using Agrobacterium tumefaciens and phenotypic variation of transformed plants
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Transformation of Solanum tuberosum L. with potato leafroll virus (PLRV) coat protein gene using Agrobacterium tumefaciens and phenotypic variation of transformed plants

机译：农杆菌介导的马铃薯卷叶病毒（PLRV）外壳蛋白基因转化马铃薯的研究及表型变异

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摘要

Leaf segments of the United States genotypes, including established cultivars (Norchip, Russet Norkotah, Goldrush, and Norqueen Russet) and a breeding line (ND860-2) of potato (Solanum tuberosum) were inoculated with Agrobacterium tumefaciens that carried a binary vector, containing the coat protein (CP) gene of potato leafroll virus (PLRV). After inoculation, calluses and shoots were formed from leaf segments in the presence of kanamycin. Regenerated shoots were transferred to rooting media whichcontained higher levels of kanamycin to select against escapes. The genotypes of ND860-2 and Norchip, were easily transformed and regenerated into shoots in contrast to the delayed and low frequencies of transformation in Russet Norkotah, Goldrush, and Norqueen Russet. The presence of the CP gene was verified by dot blot hybridization and the gene copy number was analysed via Southern hybridization. Coat protein of PLRV in transgenic plants was not detected with enzyme-linked immunosorbent assay but wasdetected with western analysis. Transformed plants appeared phenotypically normal, except for one clone that showed a light yellow leaf colour. The white-skinned genotypes had higher frequencies of transformation and regeneration than the russet-skinnedgenotypes.
机译：用携带二元载体的根癌农杆菌接种美国基因型的叶段，包括成熟的品种（Norchip，Russet Norkotah，Goldrush和Norqueen Russet）和马铃薯（Solanum tuberosum）的育种系（ND860-2）。马铃薯卷叶病毒（PLRV）的外壳蛋白（CP）基因。接种后，在存在卡那霉素的情况下，由叶段形成愈伤组织和芽。将再生的芽转移到生根培养基中，所述生根培养基含有较高水平的卡那霉素以选择逃逸的植物。与Russet Norkotah，Goldrush和Norqueen Russet中转化的延迟和低频率相比，ND860-2和Norchip的基因型易于转化并再生为芽。通过斑点印迹杂交验证CP基因的存在，并通过Southern杂交分析基因拷贝数。酶联免疫吸附法未检测到转基因植物中PLRV的外壳蛋白，而Western分析则检测到。转化的植物在表型上看似正常，除了一个克隆显示出淡黄色的叶色。白皮肤基因型的转化和再生频率高于赤褐色斯基恩奇型。

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