带npt-Ⅱ基因转基因水稻快速检测技术建立.pdfVIP

HEREDITAS (Beijing) 2007 4 , 29(4): 499507 ISSN 0253-9772 DOI: 10.1360/yc-007-0499 npt- , , , , , , , 410128 : 9 (ZH9(R)) 9 (ZH9(CK)), ZH9(R) npt- , , npt- (Kanamycin, Kan)G418 ZH9(R)ZH9(CK) , , G418 npt- , G418( )80 mg/L (4 ) G418 ZH9(R)ZH9(CK) : (1) G418()300 mg/L (7 ) ; (2) G418(1/2 MS+0.5 mg/L 6-BA+1.5%)200 mg/L (10 ) ; (3) G418(1/2 MS+0.5 mg/L 6-BA+1.5%)150 mg/L ( 12 ), PCR ZH9(R), npt- : ; npt- ; G418; Quick testing-technology of transgenic rice with npt- screen marker gene WANG Zi-Xuan, YI Zi-Li, WANG Zhi-Cheng, JIANG Jian-Xiong, QIN Jing-Ping, ZHANG Hao, TAN Yan-Ning Key Laboratory of Cell Engineering, Hunan Agricultural University, Changsha 410128, China Abstract: By using npt- gene as assistant selection-marker, treating Japonica rice Zhonghua 9 [ZH9 (CK)] and trans- ferred lysozyme gene rice (the donor rice is Japonica rice Zhonghua 9) [ZH9(R)] with antibiotics, we built a system of quickly testing transgenic rice offspring. Detached leaves of ZH9(R) and ZH9(CK) were treated by Kanamycin (0, 400, 450, 500, 550, 600 and 700 mg/L) and G418 (0, 40, 60, 80, 100, 150 and 200 mg/L) with different concentrations. The result show effect of Kanamycin is not evident and G418 is the best antibiotics to test transgenic rice with npt- gene. The data showed that 80 mg/L G418 (treating 4 d) was optimal to test transgenic rice. Further study was done with seeds, young em- bryos and seedlings by G418 testing: the alive seeds were cultured in the culture dishes which filled with a series of G418 solution with different concentration of 0, 100, 150, 200, 250, 30