【作者】 李凤娟；

【导师】 姜廷波；

【作者基本信息】 东北林业大学 ， 林木遗传育种， 2007， 硕士

【摘要】 本研究利用反义RNA技术验证植物铁蛋白的基因功能。用农杆菌介导法将大豆铁蛋白基因SoyFer1 （M 64337）和烟草内源反义铁蛋白基因NtFer2 （AY 141105）的cDNA分别导入烟草，得到转基因烟草，并采集烟草种子。对T₁转基因烟草的卡那霉素抗性分析表明，整合到烟草基因组的外源基因多为单拷贝基因，也有少数为多拷贝基因。对具有卡那霉素抗性的转基因植株进行PCR-Southern检测和Northern杂交分析表明，外源基因已整合到烟草基因组，并且得到了正确表达。将转基因株系移栽到铁离子不同的培养基中生长进行比较表明，转大豆铁蛋白基因株系的生长量明显高于非转基因株系和转内源反向铁蛋白基因烟草株系，而转内源反义铁蛋白基因株系的生长量则明显低于与非转基因株系和转大豆铁蛋白基因株系。转大豆铁蛋白基因株系和转内源反义铁蛋白基因株系的叶绿素量、MDA含量和POD活性等生理性状也发生了明显变化。表现为转大豆铁蛋白基因株系的叶绿素量和POD活性明显高于与非转基因株系，而转内源反义铁蛋白基因株系的叶绿素量和POD活性等则明显低于与非转基因株系。运用同样的方法将大豆铁蛋白基因SoyFer1导入杨树基因组中，对转基因杨树进行了PCR、PCR-Southern检测，初步证明SoyFer1整合到杨树基因组中。更多还原

【Abstract】 For studying the effects of ferritin gene overexpression and interference on grow rate intransgenic tobacco （Nicotiana tabacum L.） plants expressing soybean （Glycine max Merr） ferritingene （SoyFerl, M 64337） and antisense tobacco ferritin gene （NtFer2, AY 141105）, theexpression vector has been constructed by placing soybean ferritin cDNA and antisense ferritincDNA under the control of the CaMV 35S promoter through replace GUS gene of pBI121,transgenic tobacco were produced by co-cultivation of leaf disks with Agrobacteriumtumefaciens containing the vector. Kanamycin resistance analysis of T₁ generation of transgenictobacco shows that most of T₁ generation has 3:1 separate ratios, only few has 15:1 separateratios. This indicated that about 1-2 copy exogenous genes intergraded into tobacco genome. Theexogenous gene expression was examined by Northern-blot analyses. Comparison of the growthrate between nontransformant and transgenic tobacco plants grown on MS medium containinghigh Fe²⁺, the enhanced growth of transgenic tobacco expressing soybean ferritin gene wasobserved at the early development stages, resulting in plant height and fresh weights significantlygreater than those in the nontransformants and transformants expressing ant-tobacco ferritin gene.Soybean ferritin gene SoyFerl was transferred into P.davidiana×P.bolleana by the same way withTobacco.更多还原