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为进一步明确甜瓜果实软化的分子机理,构建了 ACC 氧化酶Ⅰ基因(CmACOⅠ)启动子与GUS 基因融合的植物表达载体,采用根癌农杆菌介导法转化甜瓜‘甘甜一号’。通过卡那霉素抗性和 PCR检测筛选呈阳性的转化植株,取不同组织进行 X-Gluc 染色。结果表明,转化植株的根、茎、叶、花、果实等器官组织经 X-Gluc 染色后,只在花药组织和成熟果果皮中出现蓝色斑点,其余组织均未检出,表明甜瓜 CmACOⅠ启动子能够驱动 GUS 基因在转基因甜瓜花药和成熟果果皮中特异表达。
In order to further clarify the molecular mechanism of melon fruit softening, we constructed a plant expression vector fused with the promoter of ACC oxidase Ⅰ gene (CmACO Ⅰ) and GUS gene and transformed melon ’Sweet No.1’ by Agrobacterium tumefaciens. The positive transgenic plants were screened by kanamycin resistance and PCR, and different tissues were taken for X-Gluc staining. The results showed that, after X-Gluc staining, the tissues of roots, stems, leaves, flowers, fruits and other organs of the transformed plants showed blue spots only in the anther and mature peel, and the rest tissues were not detected, indicating that CmACOⅠ Subgenus can drive the GUS gene to be specifically expressed in the transgenic melon anthers and mature fruit peels.