玉米转录因子Pl表达载体的构建及大花君子兰遗传转化体系的建立

The Construction of Pl Gene Expression Vector in Maize and Genetic Transformation System in Clivia miniata

大花君子兰是具有广泛市场价值的重要室内观赏花卉。但是,常规遗传育种周期时间长,污染率高和转化率低等原因,是君子兰遗传育种中普遍存在的问题。针对这些问题,本研究构建了以甘露糖异构酶基因(Pmi)为筛选基因并带有玉米Pl转录因子的pBI121-Pl真核表达载体,以茎尖分生组织作为受体,进行了大花君子兰遗传转化体系的探究。确定了无菌植株的最佳遗传转化方案、准确的共培养时间和温度以及最适甘露糖筛选浓度,初步证明Pl基因已成功整合到大花君子兰基因组中并获得了转基因植株,有助于君子兰的遗传育种的研究。

Clivia miniata is an important indoor ornamental flower which has been reported to have broad market value. But there are lots of problems existing in the breeding process of Clivia miniata, such as the long cycle,higher pollution rate and lower conversion rate. In order to solve these problems, we successfully constructed expression vector with resistance gene phosphomannose isomerase（Pmi） and maize Pl transcription factor, used stem-tip tissue as acceptor and studied the genetic transformation system of Clivia miniata. We confirmed the optimum transformation scheme, precise co-culture time and temperature, and proper concentration of mannose.Finally, we obtained the transgenic plants ensured by mannose selection and PCR detection, which were beneficial to the genetic breeding of Clivia miniata.