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为提高苦参毛状根诱导率,探索苦参总碱生产的新资源和途径。分别用不同浓度的发根农杆菌R1601转化苦参诱导毛状根,用发根农杆菌R1601侵染苦参后和不同浓度的2,4二氯苯氧乙酸(2,4-D)和6-苄氨基嘌呤(6-BA)共培养诱导毛状根,利用PCR方法检测脱菌完全的毛状根,对获得的毛状根株系继代培养,并对苦参毛状根的苦参总碱含量进行了测定。结果:发根农杆菌R1601菌液浓度在吸收值(A600)为0.8时最适合毛状根的诱导;添加0.375 mg/L的2,4-D和0.375 mg/L的6-BA时,诱导率最高,可达56.7%;PCR鉴定表明,诱导的毛状根含TL-DNA序列;毛状根株系中苦参总碱含量比2年生苦参根中苦参总碱含量略低。表明利用苦参毛状根大规模培养可以获得苦参总碱。
In order to improve the induction rate of hairy root of Sophora flavescens, explore the new resources and ways of the production of Sophora alkaloids. The roots of A. mandshurica were transformed with Agrobacterium rhizogenes R1601 at different concentrations, respectively, and the infected roots were infected with Agrobacterium rhizogenes R1601 and 2,4 dichlorophenoxyacetic acid (2,4-D) and 6 - benzylaminopurine (6-BA) were used to induce the hairy roots. The decellularized hairy roots were detected by PCR. The hairy roots were subcultured, The alkali content was determined. Results: The optimum concentration of Agrobacterium rhizogenes R1601 was induced by hairy roots at the absorbency (A600) of 0.8. When 0.375 mg / L 2,4-D and 0.375 mg / L 6-BA were added, The highest rate was 56.7%. PCR identification showed that induced hairy roots contained TL-DNA sequence. Matrine content in hairy roots was slightly lower than that in 2-year-old Sophora flavescens roots. Show that the use of mandarin hairy roots large-scale culture can get matrine alkaloids.