发根农杆菌转染纹党产生毛状根试验
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摘要
试验采用正交试验法探究不同OD值、侵染时间及外植体类型对诱导纹党产生毛状根的最佳条件,结果表明,采用OD值0.8A、侵染时间10min与纹党幼嫩叶片作为外植体的试验组处理效果最好,毛状根诱导率可达55.6%。纹党幼嫩叶片作为外植体诱导毛状根处理效果最好,纹党根、纹党茎段及无菌苗皆不适宜作为诱导材料。经形态学观察,诱导出的根基本符合毛状根的特征,PCR扩增得到865bp的DNA目的条带,证明试验得到的根为发根农杆菌R1601侵染诱导得到的毛状根。
In this experiment,the orthogonal test method was used to explore different OD values,infection time and explant type to optimize the hairy roots of the induced striate. The experiment showed that the OD value was 0.8 A,the infection time was 10 min and the pattern was young. The test group of the tender leaves as the explants had the best treatment effect,and the hairy root induction rate reached 55.6%. The young leaves of the striatum were the bestfor the treatment of hairy roots as explants,and the stalk roots,the stem segments and the sterile seedlings were not suitable as inducing materials.The 865 bp DNA target band was obtained by morphological observation and PCR amplification. The root obtained in this experiment was confirmed to be the hairy root induced by Agrobacterium rhizogenes R1601 infection.
In this experiment,the orthogonal test method was used to explore different OD values,infection time and explant type to optimize the hairy roots of the induced striate. The experiment showed that the OD value was 0.8 A,the infection time was 10 min and the pattern was young. The test group of the tender leaves as the explants had the best treatment effect,and the hairy root induction rate reached 55.6%. The young leaves of the striatum were the bestfor the treatment of hairy roots as explants,and the stalk roots,the stem segments and the sterile seedlings were not suitable as inducing materials.The 865 bp DNA target band was obtained by morphological observation and PCR amplification. The root obtained in this experiment was confirmed to be the hairy root induced by Agrobacterium rhizogenes R1601 infection.
引文
[1]聂玲霞,张双定.陇西县党参栽培技术规程[J].甘肃农业科技,2016,5(10):21-25.
[2]王洁,邓长泉,石磊,等.党参的现代研究进展[J].中国医药指南,2011,09(31):279-281.
[3]王伟,陆杨,李礼,等.喜树毛状根的诱导及其喜树碱含量分析[J].西北植物学报,2008,28(12):2416-2422.
[4]刘树楠,孙天恩,李根保.银杏发根的转化及其悬浮培养无性系的建立[J].武汉大学学报(理学版),1998(2):238-240.
[5]杨蕊,张家辉,孙雪梅,等.杜仲毛状根培养条件优化及其桃叶珊瑚苷药用成分的研究[J].中国中药杂志,2009,34(15):1902-1905.
[6]黄素梅,石丸幹二.乌桕毛状根培养与次生代谢产物累积研究[J].北方园艺,2011(22):103-105.
[7]Jia Z,Koike K,Nikaido T,et al.Triterpenoid saponins from Ardisia crenata and their inhibitory activity on c AMP phosphodiesterase[J].Chemical&Pharmaceutical Bulletin,1994,42(11):2309-2314.
[8]任如意,李思璇,金秋,等.发根农杆菌诱导毛酸浆毛状根条件的优化[J].北方园艺,2017(8):115-119.
[9]Slightom JL,Durand-Tardif M,Jouanin L,et al.Nucleotidesequence analysis of TL-DNA of Agrobacterium rhizogenesagropine type plasmid.Identification of open reading frames[J].Journal of Biological Cemistry,1986,261(1):108-121.
[10]姚庆收,陈向明,马玉娥.蒙古黄芪毛状根的高效诱导及毛状根生物量和总黄酮含量的比较分析[J].中药材,2017.
[2]王洁,邓长泉,石磊,等.党参的现代研究进展[J].中国医药指南,2011,09(31):279-281.
[3]王伟,陆杨,李礼,等.喜树毛状根的诱导及其喜树碱含量分析[J].西北植物学报,2008,28(12):2416-2422.
[4]刘树楠,孙天恩,李根保.银杏发根的转化及其悬浮培养无性系的建立[J].武汉大学学报(理学版),1998(2):238-240.
[5]杨蕊,张家辉,孙雪梅,等.杜仲毛状根培养条件优化及其桃叶珊瑚苷药用成分的研究[J].中国中药杂志,2009,34(15):1902-1905.
[6]黄素梅,石丸幹二.乌桕毛状根培养与次生代谢产物累积研究[J].北方园艺,2011(22):103-105.
[7]Jia Z,Koike K,Nikaido T,et al.Triterpenoid saponins from Ardisia crenata and their inhibitory activity on c AMP phosphodiesterase[J].Chemical&Pharmaceutical Bulletin,1994,42(11):2309-2314.
[8]任如意,李思璇,金秋,等.发根农杆菌诱导毛酸浆毛状根条件的优化[J].北方园艺,2017(8):115-119.
[9]Slightom JL,Durand-Tardif M,Jouanin L,et al.Nucleotidesequence analysis of TL-DNA of Agrobacterium rhizogenesagropine type plasmid.Identification of open reading frames[J].Journal of Biological Cemistry,1986,261(1):108-121.
[10]姚庆收,陈向明,马玉娥.蒙古黄芪毛状根的高效诱导及毛状根生物量和总黄酮含量的比较分析[J].中药材,2017.