Nuñez-Palenius Hector G, Cantliffe Daniel J, Huber Don J, Ciardi Joseph, Klee Harry J
Horticultural Sciences Department, University of Florida, IFAS, Gainesville, FL 32611, USA.
Plant Cell Rep. 2006 Mar;25(3):198-205. doi: 10.1007/s00299-005-0042-0. Epub 2005 Dec 16.
'Galia' muskmelon (Cucumis melo L. var. reticulatus Ser.) has been recalcitrant to transformation by Agrobacterium tumefaciens. Transformation of the 'Galia' male parental line, 'Krymka', with an ACC oxidase (CMACO-1) gene in antisense orientation is described herein. Explants were transformed using A. tumefaciens strain ABI, which contained a vector pCmACO1-AS plasmid, bearing an antisense gene of CMACO-1 and the CP4 syn gene (glyphosate-tolerance). Both CMACO-1 and CP4 syn genes were assessed by a polymerase chain reaction method. Flow cytometry analysis was performed to determine plant ploidy level of primary transformants. Two completely diploid independent transgenic plants were obtained. Southern blot and segregation analysis in the T1 generation determined that each independent transgenic line had one single insertion of the transgene. These transgenic muskmelon male parental lines have potential for use in the production of 'Galia' F1 hybrids with improved shelf life.
“加利娅”甜瓜(Cucumis melo L. var. reticulatus Ser.)一直难以通过根癌农杆菌进行转化。本文描述了将反义方向的ACC氧化酶(CMACO - 1)基因转化到“加利娅”雄性亲本系“克里姆卡”中的过程。外植体使用根癌农杆菌菌株ABI进行转化,该菌株含有载体pCmACO1 - AS质粒,带有CMACO - 1的反义基因和CP4 syn基因(耐草甘膦)。通过聚合酶链反应方法对CMACO - 1和CP4 syn基因进行评估。进行流式细胞术分析以确定初级转化体的植物倍性水平。获得了两个完全二倍体的独立转基因植株。Southern印迹分析和T1代的分离分析确定每个独立的转基因系都有转基因的单一插入。这些转基因甜瓜雄性亲本系有潜力用于生产货架期延长的“加利娅”F1杂种。