Construction of Antisense ACC Oxidase Gene of Lilium and its Genetic Transformation

in HortScience
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  • 1 Key Laboratory of Northwest Horticulture Plant Germplasm and Genetic Improvement of Ministry of Agriculture, Northwest A&F University, College of Horticulture, Yangling, Shaanxi, 712100, P.R. China

The enzyme ACC oxidase (ACO), encoded by a small multigene family in many plants, catalyzes the terminal step in the ethylene biosynthesis pathway. In this research, based on the total RNA isolated from the flowers of Asia hybrids `Pollyanna' and Oriental hybrids `Sorbonne', we obtained two cDNA fragments of ACO genes (Genbank accession DQ062133 and DQ062134) by RT-PCR technique. The two cDNA fragments were reversely inserted into plant expression vector pWR306 respectively, and constructed two antisense ACO gene expression binary vectors harboring hygromycin phosphotransferase (hptII), glucuronidase (uid A), and a green fluorescent protein (GFP) gene in the T-DNA region. We have developed a system to produce transgenic plants in LiLium via Agrobacterium tumefaciens-mediated transformation of calli. Transformants were subjected to GFP expression analysis, PCR assay, and Southern hybridization to confirm gene integration.

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