Yu Yi-Xun, Bao Man-Zhu
Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China.
Sheng Wu Gong Cheng Xue Bao. 2004 Sep;20(5):704-7.
The cultivar 'Master' of carnation (Dianthus caryophyllus L.) was transformed with four T-DNA structures containing sense, antisense, sense direct repeat and antisense direct repeat gene of ACC oxidase mediated by Agrobacterium tumefaciens. Southern blotting detection showed that foreign gene was integrated into the carnation genome and 14 transgenic lines were obtained. The transgenic plants were transplanted to soil and grew normally in greenhouse. Of the 12 transgenic lines screened, the cut flower vase life of 8 transgenic lines is up to 11 days and the longest one is 12.8 days while the vase life of the control is 5.8 days under 25 degrees C. The vase life of 2 lines out of 3 with single sense ACO gene is same as that of the control, while the vase life of 3 lines out of 4 with single antisense ACO gene is prolonged. The vase life of cut flowers of 5 lines with direct repeat ACO genes is all prolonged by about 6 days, while the vase life of 3 out of 7 lines with single ACO gene is same as that of the control. During the senescence of cut flowers, the ethylene production of the most of the transgenic lines decreased significantly, and the production of ethylene is not detectable in lines T456, T556 and T575. The results of the research demonstrate that antisense foreign gene inhibits expression of endogenesis gene more significantly than sense one. Both sense direct repeat and antisense direct repeat foreign genes can suppress endogenous gene expression more significantly comparing to single foreign genes. The transgenic lines obtained from this research are useful to minimize carnation cut flower transportation and storage expenses.
以农杆菌介导,将含有ACC氧化酶基因的正义、反义、正义同向重复和反义同向重复4种T-DNA结构导入香石竹(Dianthus caryophyllus L.)品种‘Master’中。Southern杂交检测表明,外源基因已整合到香石竹基因组中,共获得14个转基因株系。将转基因植株移栽到土壤中,在温室中正常生长。在筛选出的12个转基因株系中,25℃条件下,8个转基因株系的切花瓶插寿命长达11天,最长的为12.8天,而对照的瓶插寿命为5.8天。3个含单个正义ACO基因的株系中有2个的瓶插寿命与对照相同,4个含单个反义ACO基因的株系中有3个的瓶插寿命延长。5个含同向重复ACO基因的株系切花瓶插寿命均延长约6天,7个含单个ACO基因的株系中有3个的瓶插寿命与对照相同。在切花衰老过程中,大多数转基因株系的乙烯释放量显著降低,T456、T556和T575株系检测不到乙烯释放。研究结果表明,反义外源基因比正义外源基因更能显著抑制内源基因的表达。与单个外源基因相比,正义同向重复和反义同向重复外源基因均能更显著地抑制内源基因表达。本研究获得的转基因株系有助于最大限度地降低香石竹切花的运输和贮藏成本。