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当前位置: 首页 > 文章 > 以PMI为选择标记的露地菊转Lc-14-3-3基因体系的建立及功能鉴定 园艺学报 2011,38 (6) 1139-1146
Position: Home > Articles > Phosphomannose Isomerase/Mannose Selection System to Screen Lc14-3-3 Transgenic Chrysanthemum Plants Acta Horticulturae Sinica 2011,38 (6) 1139-1146

以PMI为选择标记的露地菊转Lc-14-3-3基因体系的建立及功能鉴定

作  者：
于利刚;解莉楠;王江;张旸;李玉花
单  位：
东北林业大学生命科学学院
关键词：
菊;露地菊;Lc14-3-3;磷酸甘露糖异构酶基因;转基因
摘  要：
从东北地区高pH盐碱地上生长良好的羊草中克隆Lc14-3-3基因,以露地菊(Chrysanthemum morifolium)‘火焰’叶片愈伤组织为受体,以农杆菌介导,将Lc14-3-3基因转入‘火焰’基因组中。以载体中的磷酸甘露糖异构酶基因(phosphomannose isomerase,PMI)为筛选标记,通过甘露糖的筛选,结合PCR、Southern杂交检测Lc14-3-3整合到其基因组。通过转基因植株与对照植株的耐盐性对比可见转化Lc14-3-3基因植株具有更高的耐盐能力。磷酸甘露糖异构酶基因/甘露糖筛选系统可有效应用于露地菊遗传转化。
译  名：
Phosphomannose Isomerase/Mannose Selection System to Screen Lc14-3-3 Transgenic Chrysanthemum Plants
作  者：
YU Li-gang,XIE Li-nan,WANG Jiang,ZHANG Yang,and LI Yu-hua(College of Life Science,Northeast Forestry University,Harbin 150040,China)
关键词：
chrysanthemum;Lc14-3-3;PMI;transgenic
摘  要：
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.

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