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High-efficiency and stable genetic transformation of pear (Pyrus communis L.) leaf segments and regeneration of transgenic plants

文献类型： 外文期刊

作者： Sun, Qingrong 1 ; Zhao, Yan 1 ; Sun, Hongyan 1 ; Hammond, Rose W. 1 ; Davis, Robert E. 1 ; Xin, Li 2 ;

作者机构： 1.ARS, Mol Plant Pathol Lab, USDA, Beltsville, MD 20705 USA

2.Shandong Inst Pomol, Tainan 271000, Shandong, Taiwan

关键词： genomic DNA;transgene;T-DNA;disease resistance;transformation induction strategy

期刊名称：ACTA PHYSIOLOGIAE PLANTARUM （ 影响因子：2.354； 五年影响因子：2.711 ）

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收录情况： SCI

摘要： Pear (Pyrus communis L.) is a nutrient-dense fruit with strong consumer demand and high commercial value. However, most cultivated pear varieties are often susceptible to diseases caused by fungi, bacteria, and viruses. The purpose of the present study was to establish an efficient genetic transformation and regeneration protocol, paving the way for genetic engineering of pear cultivars with enhanced disease resistance. Major factors that influence transformation and regeneration were examined and optimal conditions were established for efficient transformation from leaf explants of 'Old Home', a valuable pear interstem and rootstock. High transformation efficiency was achieved largely due to an improved infection/transformation induction strategy. Co-cultivation of Agrobacterium cells and leaf segments on a liquid induction medium yielded a fivefold increase in transformation frequency. Southern hybridization analysis revealed presence of reporter gene uidA in the genomic DNA samples from independent transgenic plants, confirming the integration of the transgene in recipient pear genomes. The stability of T-DNA integration was evaluated by the consistent presence of the Km selectable marker and the expression pattern of the introduced reporter gene uidA was analyzed by GUS histochemical assay.

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