1.林木遗传育种全国重点实验室/林木育种与生态修复国家工程研究中心/北京林业大学生物科学与技术学院,北京 100083
2.内蒙古自治区赤峰市敖汉旗双井林场,赤峰 024300
马灵,E-mail:lingmaml@163.com
庞晓明,E-mail:xmpang@163.com
收稿:2024-04-12,
纸质出版:2025-01-15
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MA Ling,YUAN Yi,YANG Shuang,et al.Establishment of genetic transformation system for leaves of ‘Jingzao 39’(Ziziphus jujuba Mill.)[J].Journal of Huazhong Agricultural University,2025,44(01):43-49.
马灵,袁轶,杨爽,等.‘京枣39’叶片遗传转化体系的建立[J].华中农业大学学报,2025,44(1):43‐49. DOI: 10.13300/j.cnki.hnlkxb.2025.01.005.
MA Ling,YUAN Yi,YANG Shuang,et al.Establishment of genetic transformation system for leaves of ‘Jingzao 39’(Ziziphus jujuba Mill.)[J].Journal of Huazhong Agricultural University,2025,44(01):43-49. DOI: 10.13300/j.cnki.hnlkxb.2025.01.005.
为建立‘京枣39’高效遗传转化体系,以‘京枣39’叶片为外植体进行遗传转化,探究卡那霉素(kanamycin,Kan)质量浓度、头孢噻肟(cefotaxime, Cef)和特美汀(timentin, Tim)抑菌质量浓度、预培养时间、不同菌液浓度、侵染时间、乙酰丁香酮(acetosyringone, AS)浓度及共培养时间对‘京枣39’叶片转化率的影响。结果显示,‘京枣39’叶片最佳遗传转化条件为,5 mg/L Kan选择压,50 mg/L Cef、250 mg/L Tim抑菌质量浓度,预培养4 d,农杆菌菌液OD
600 nm
值为0.6,侵染时间20 min,共培养基添加200 μmol/L AS,共培养4 d。荧光检测和PCR试验证实外源目的基因已成功整合到‘京枣39’基因组中,实现了农杆菌介导的‘京枣39’叶片的遗传转化。
Leaves of ‘Jingzao 39’were used as explants to establish an efficient genetic transformation system for ‘Jingzao 39’. The effects of the selection pressure of kanamycin (Kan), the inhibitory concentration of cefotaxime (Cef) and timentin (Tim), pre-culture time, the density of bacterial suspension, infection duration, the concentration and co-culture time of acetylyringone (AS) on the transformation rate of leaves of ‘Jingzao 39’ were studied. The results showed that the optimal genetic transformation conditions for the leaves of ‘Jingzao 39’ were as follows: 5 mg/L Kan, 50 mg/L Cef, 250 mg/L Tim, pre-culture for 4 days,
Agrobacterium tumefaciens
concentration OD
600 nm
=0.6, infection duration of 20 minutes, 200 μmol/L AS, co-culture for 4 days. The results of fluorescence detection and PCR analyses confirmed that the exogenous target gene was successfully integrated into the genome of ‘Jin
gzao 39’, achieving the
Agrobacterium
mediated genetic transformation of leaves of ‘Jingzao 39’. It will provide an important support for analyzing the function of genes and molecular breeding in jujube.
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