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Production of dwarf lettuce by overexpressing a pumpkin gibberellin20-oxidase gene

机译:通过过表达南瓜赤霉素20-氧化酶基因生产矮生菜

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We investigated the effect of overexpressing a pumpkin gibberellin (GA) 20-oxidase gene encoding an enzyme that forms predominantly biologically inactive products on GA biosynthesis and plant morphology in transgenic lettuce (Lactuca sativa cv Vanguard) plants. Lettuce was transformed with the pumpkin GA 20-oxidase gene downstream of a strong constitutive promoter cassette (E12-35S-Omega). The transgenic plants in which the pumpkin gene was detected by polymerase chain reaction were dwarfed in the T-2 generation, whereas transformants with a normal growth phenotype did not contain the transgene. The result of Southern-blot analysis showed that the transgene was integrated as a single copy; the plants segregated three dwarfs to one normal in the T-2 generation, indicating that the transgene was stable and dominant. The endogenous levels of GA(1) and GA(4) were reduced in the dwarfs, whereas large amounts of GA(17) and GA(25), which are inactive products of the pumpkin GA 20-oxidase, accumulated in these lines. These results indicate that a functional pumpkin GA 20-oxidase is expressed in the transgenic lettuce, resulting in a diversion of the normal pathway of GA biosynthesis to inactive products. Furthermore, this technique may be useful for controlling plant stature in other agricultural and horticultural species.
机译:我们调查了过表达南瓜赤霉素(GA)20-氧化酶基因的作用,该基因编码一种在转基因生菜(Lactuca sativa cv Vanguard)植物中主要形成生物惰性产物的酶,对GA生物合成和植物形态具有影响。用强组成型启动子盒(E12-35S-Omega)下游的南瓜GA 20-氧化酶基因转化生菜。通过聚合酶链反应检测到南瓜基因的转基因植物在T-2世代中相形见,,而具有正常生长表型的转化体不包含该转基因。 Southern印迹分析的结果表明,转基因被整合为单个拷贝。这些植物在T-2世代中将3个矮人隔离为1个正常人,这表明转基因是稳定且占优势的。侏儒体内的内源性GA(1)和GA(4)含量降低,而南瓜GA 20氧化酶的非活性产物大量GA(17)和GA(25)积累在这些品系中。这些结果表明,功能性南瓜GA 20-氧化酶在转基因生菜中表达,导致GA生物合成的正常途径转移至无活性产物。此外,该技术对于控制其他农业和园艺物种中的植物高度可能是有用的。

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