【摘要】 利用农杆菌介导方法,分别将PAPⅡ单基因、PAPⅡ基因和异戊烯基转移酶基因(ipt)共表达的双基因(PAPⅡ-ipt)导入烟草品种K326叶细胞中,半定量RT-PCR分析了转基因植株中PAPⅡ、ipt和核糖体蛋白大亚基基因(RPL3A)的表达,并进行了TMV攻毒实验。结果表明:双基因PAPⅡ-ipt对烟草的转化频率高达45.55%,为单基因PAPⅡ的转化频率(13.3%)的3.4倍;50%的转基因植株没有检测到转基因的表达,PAPⅡ-ipt表达的转基因植株中RPL3A基因的表达水平为PAPⅡ表达植株中的1.25倍,且生长发育正常,对TMV病毒的抗性有较大程度的提高。因此,ipt与PAPs基因共表达可作为克服PAPs细胞毒性的有效途径。更多还原

【Abstract】 The gene PAPⅡand the co-expressed genes of PAPⅡ-ipt(isopentenyl transferase) were introduced into mesophyll cells of Nicotiana tabacum L.var.K326,respectively,mediated by Agrobacterium tumefaciens.The expression levels of the genes such as PAPⅡ,ipt and RPL3 A(Ribosomal Protein Li) in transgenic plants were determined by semi-quantitative RT-PCR,and the TMV challenge experiment was conducted for the transgenic plants.The results showed that transformation frequency of the double genes PAPⅡ-ipt reached 45.55%,3.4 times as much as that of PAPⅡgene alone,transcription of the trans-genes was not determined in 50%of the transgenic plants,and the expression level of RPL3A in transgenic plants with expression of the double genes was 1.25 times as much as that in transgenic plants with expression of the single PAPⅡgene.Moreover,the transgenic plants with co-expression of the PAPⅡand ipt genes had normal phenotype and fertility,as well as exhibited higher degree of resistance to the TMV.Thus,it might be suggested that the co-expression of ipt and PAPⅡgenes is an effective solution to reduce PAPs cytotocixity.更多还原