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为研究山茶花CjAPL1基因对于重瓣花形成的作用,构建了pCAMBIA1300-CjAPL1正义植物表达载体,酶切结果显示,CjAPL1基因正向插入pCAMBIA1300的启动子和终止子之间,表明CjAPL1植物表达载体构建成功。将pCAMBIA1300-CjAPL1采用农杆菌介导的花序浸染法转化拟南芥,获得转基因阳性植株65株。随机挑选表型变异明显的3株进行PCR扩增都得到了目的条带,Southern杂交鉴定进一步确认为转基因阳性植株。同时荧光定量检测到在转基因植株中CjAPL1基因表现出较对照显著提高6~25倍的表达量。转基因阳性植株表型变异明显,花柱和雄蕊数相比野生型各增加1枚和1~4枚,萼片边缘发育成白色的瓣化状,表明在拟南芥中过量表达CjAPL1基因可以引起花器官形态的变异和数量的变化,因此该基因在花器官形成和重瓣花发育中具有显著的调控功能。
In order to study the effect of CjAPL1 gene on the formation of double flower in Camellia japonica, a positive expression vector pCAMBIA1300-CjAPL1 was constructed. The results of digestion showed that the positive CjAPL1 gene was inserted between the promoter and terminator of pCAMBIA1300, indicating that CjAPL1 plant expression vector was successfully constructed . Arabidopsis thaliana was transformed into pCAMBIA1300-CjAPL1 by Agrobacterium-mediated inflorescence, and 65 transgenic plants were obtained. Three bands with obvious phenotypic variation were randomly selected for PCR amplification. The Southern blotting was further confirmed as transgenic positive plants. At the same time, fluorescence quantitative detection of CjAPL1 gene in transgenic plants showed a significant increase 6 to 25 times the expression of the control. Transgenic plants showed obvious phenotypic variation. The numbers of styles and stamens increased by 1 and 1 ~ 4 respectively compared with the wild type. The edges of sepals developed white petals, which indicated that overexpression of CjAPL1 gene in Arabidopsis could cause flower organs Morphological variation and quantitative changes, so the gene has a significant regulatory function in floral organ formation and development of double flowering flowers.