关键词: 菊, 露地菊, Lc14-3-3, 磷酸甘露糖异构酶基因, 转基因

Abstract: The objective of this experiment was to develop the selection system for transgenic chrysanthemums（Chrysanthemum morifolium）by Agrobacterim tumefaciens carrying genes related in saline-alkaline tolerance without using antibiotic resistance genes. Lc14-3-3 was cloned from a xerophilous grass，Leymus chinensis，which is adapted to high pH（8.5–11.5）sodic soils. Leaf explants from in vitro germinated plants of chrysanthemum‘Huoyan’were infected with A. tumefaciens EHA105 pCAMBIA1301. The selection system was based on the use of Escherichia coli phosphomannose isomerase（PMI）gene as a selectable marker gene and mannose as a selective nutrient. The explants were selected on medium supplemented with a combination of mannose and sucrose as a carbon source at different concentrations. Among the leaf explants which were infected，there are 19 shoots surviving after selection by the medium containing 15 g · L-1 mannose. PCR analysis and southern blot analysis confirmed that T3 independent transformed lines were positive for PMI and Lc14-3-3 gene. The growth of transformed lines showed more healthy and high tolerant. The PMI/mannose selection system was efficient for producing transgenic chrysanthemum plants without using antibiotics or herbicides.

Key words: chrysanthemum, Lc14-3-3, PMI, transgenic