转il-4基因番茄及其杂交F_1代的筛选与表现型分析

Selection and Phenotypic Analysis of Transgenic Tomato with Interleukin-4 Gene and its F_1 hybrids

为完善转il-4基因番茄的筛选及鉴定工作,本实验比较了il-4基因在PPT抗性筛选与非PPT抗性筛选植株中的阳性率,对il-4基因在T6代及杂交F1代番茄中的传递规律和植株表现型进行调查。本文对L402、‘中疏4号’、‘大黄’三个品种的T6代及杂交F1代植株进行PPT抗性筛选及非PPT抗性筛选,特异性的PCR扩增、PCR-Southern杂交。研究结果表明,il-4基因在非PPT抗性筛选植株中的阳性率高于PPT筛选植株,T6代及F1代植株中il-4基因的分离情况不符合孟德尔分离规律,其分离情况远远低于3:1的分离比率。转il-4基因番茄与非转基因植株(对照)相比发现,转基因植株在一至多个性状上发生了不同程度的变异,杂交F1代与亲本T6代植株相比,杂交F1代有少数组合存在杂种优势。

In order to improve the selection and detection of transgenic tomato with interleukin-4（il-4）gene,in the present paper,positive rate ofil-4 gene in the individuals after non-PPT resistance selection was compared with those after PPT resistance selection,then the hereditary patterns and phenotype of il-4 gene of T6 transgenic individuals and F1 hybrids was analyzed.Then,DNA samples were extracted from the leaves of T6 transgenic plants and F1 hybrids of L402,＇Zhongshu4＇and＇Dahuang＇varieties,performed PPT resistance selection and non-PPT resistance selection,PCR amplification and PCR-Southern detection.The results showed that,the screening rate of target gene after non-PPT resistance selection was higher than that after PPT resistance selection.The segregation ratios ofil-4 gene of T6 and F1 individuals were much lower than 3：1,and it didn＇t conform to Mendelian inheritance.Compared to CK,variations are discovered in more than 1 character of il-4 transgenic tomato plants.A few F1 hybrids have heterosis compared to T6 transgenic individuals.