金鱼草Del基因影响烟草花色素苷的研究

Study on the Effect of Del Regulatory Gene of Antirrhinum majus on Anthocyanin of Tobacco

采用根癌农杆菌介导法，获得转Ｄｅｌ基因的转基因烟草植株．对９株可能的转基因植株（Ｔ０代）ＤＮＡ进行ＮＰＴⅡ基因的ＰＣＲ－Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交分析，其中８株显示ＮＰＴⅡ基因阳性．对其中５株开花的阳性植株进行ＤｅｌＤＮＡ的ＰＣＲ－Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交分析，全部显示ＤｅｌＤＮＡ阳性．Ｋｍ抗性Ｔ１代转基因植株ＤＮＡ进行ＤｅｌＤＮＡ的ＰＣＲ－Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交分析，结果显示全部含有ＤｅｌＤＮＡ．在５３０ｎｍ处测量Ｔ０代转基因植株中花色素苷含量和分布情况，３株转基因烟草的花萼、花冠和雄蕊部位花色素苷含量增加，而在雌蕊和叶中减少．在一定程度上验证了Ｄｅｌ基因的生物学功能，证实并补充了Ｇｏｏｄｒｉｃｈ等关于色素沉积模式的论述．

The transgenic Del gene tobacco plants were obtained by co cultivation with Agribacterium tumefacience LBA4404 (pJAM 175). Successful transformation of NPT Ⅱgene to tobacco plants of T 0 generation was demonstrated by PCR Southern blot hybridization in 8 of 9 putative transgenic plants. And then, successful transformation of Del cDNA to tobacco plants of T 0 generation was also demonstrated by PCR Southern blot hybridization in all of 5 flowering plants carrying NPT Ⅱ gene. Furthermore, the assay of PCR Southern blot hybridization demonstrated the integration of Del gene in the genome of kanamycin resistant plants of T 1 generation. The absorbance of the extracts from the transgenic plants of T 0 generation was measured at 530 nm. In 3 transgenic plants, pigment extractions from sepals, petals and stamens all accumulated more anthocyanin than the control plants. But carpels and leaves did less. To some extent, the results showed the biological functions of Del gene and demonstrated and supplemented the conclusion on pigmentation pattern mode by Goodrich et al.