摘要
利用发根农杆菌介导的遗传转化法,诱导天仙子发根.结合组织培养技术和高效液相色谱法,改变培养基碳源种类、硝态氮和铵态氮的比例和磷源的浓度,研究其对天仙子发根生长及莨菪碱和东莨菪碱质量分数的影响.结果表明:发根诱导中,C58C1的诱导效率最高,达到84.85%.蔗糖为碳源,发根干质量生物量积累最多,莨菪碱质量分数最高;果糖为碳源,东莨菪碱质量分数最高.WPM基本培养基中,保持总氮源浓度为15mmol/L,当NO_3^-与NH_4^+的比例为2∶1时,发根在生物量积累、莨菪碱和东莨菪碱质量分数上都达到最高;而当全部为NO_3^-时,发根干质量最少且莨菪碱和东莨菪碱的质量分数最低.高浓度KH_2PO_4(5.00mmol/L)时,发根的干质量最大,低浓度KH_2PO_4(0.31mmol/L)时,莨菪碱和东莨菪碱的质量分数最高.
The leaf disk transformation method mediated by Agrobacterium was performed to induce the hairy roots of Hyoscyamus niger.The hairy roots were obtained through tissue culture by manipulation of carbon,nitrogen,and phosphate sources and the contents of hyoscyamine and scopolamine were determined by HPLC.The results showed that the highest transformation efficiency of 84.85% was achieved using strain C58C1 with leaf explants.The highest dry weight of hairy roots,contents of hyoscyamine and contents of scopolamine were achieved with the addition of sucrose,maltose and fructose respectively.The achieved biomass of hairy roots,contents of hyoscyamine and scopolamine were highest when NO3-/NH4＋was 2∶1,while those were lowest in the case of NO3-as the sole N source when the concentration of total nitrogen source was kept at 15mmol/L.The highest biomass of hairy roots were achieved with the addition of high KH2PO4 concentration,while the highest contents of hyoscyamine and scopolamine were achieved with the addition of low KH2PO4 concentration.
出处
《西南大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第6期23-29,共7页
Journal of Southwest University(Natural Science Edition)
基金
教育部新世纪人才支持计划(NCET-12-0930)
国家863计划(2011AA100605)
国家自然科学基金项目(31370333)
西南大学中央高校基本科研业务费专项(XDJK2013A024
XDJK2014C064)
西南大学博士基金(SWU111012)
关键词
天仙子
发根
发根农杆菌
碳源
氮源
磷源
托品烷生物碱
Hyoscyamus niger
hairy root
carbon
nitrogen
phosphate
Agrobacterium rhizogenes
tro pane alkaloids
