College of Landscape Architecture, Sichuan Agricultural University, Chengdu 611130, China; College of Biology and Pharmacy, Yulin Normal University, Yulin 537000, China.
College of Landscape Architecture, Sichuan Agricultural University, Chengdu 611130, China.
J Biotechnol. 2020 Mar 10;311:12-18. doi: 10.1016/j.jbiotec.2020.02.009. Epub 2020 Feb 18.
To develop an alternative medicine related with taxol/camptothecin, a hairy roots induction system for measuring triterpenoid saponin ardicrenin was established. In the current study, mature and healthy seeds of Ardisia crenata plants were selected for obtaining aseptic seedlings. Two Agrobacterium rhizogenes strains ATCC 15834 and A4 were used to infect aseptic euphylla for inducing hairy roots of A. crenata plants. For the best combination of seeds germination, a Murashige-Skoog medium containing 1.0 mg L 6-benzylaminopurine and 1.0 mg L naphthalene acetic acid was made, which reached a rate of 92.4 %. Results showed that ATCC 15834 and A4 both induced hairy roots of A. crenata for improving ardicrenin production. The PCR analysis demonstrated that ATCC 15834 and A4 Ri plasmid T-DNA had been successfully transferred and integrated into the genome of leaf cell nuclei, however the Vir region was not. Further, ardicrenin content in hairy roots ACHR 15834 8.2 %) induced by ATCC 15834 was quantified by the RP-HPLC, which was also 1.8-, 2.7-, 9.4- and 2.6-fold greater than those of ACHR 4 induced by A4 (4.5 %), ACR C formed by tissue culture (3.1 %), euphylla (0.8 %) and NR C formed nature (3.2 %), respectively. Taken together, hairy root lines of A. crenata obtained were able to express naturally more ardicrenin than natural plants.
为了开发与紫杉醇/喜树碱相关的替代药物，建立了一个用于测量三萜皂苷 Ardicrenin 的发根诱导系统。在本研究中，选择成熟健康的 Ardisia crenata 植物种子获得无菌幼苗。使用两种根癌农杆菌菌株 ATCC 15834 和 A4 感染无菌 Euphylla 以诱导 A. crenata 植物的发根。为了获得最佳的种子发芽组合，制作了一种含有 1.0 mg/L 6-苄基氨基嘌呤和 1.0 mg/L 萘乙酸的 Murashige-Skoog 培养基，发芽率达到 92.4%。结果表明，ATCC 15834 和 A4 均可诱导 A. crenata 的发根，以提高 Ardicrenin 的产量。PCR 分析表明，ATCC 15834 和 A4 的 Ri 质粒 T-DNA 已成功转移并整合到叶片细胞核基因组中，但 Vir 区没有。此外，通过 RP-HPLC 定量分析由 ATCC 15834 诱导的发根 ACHR 15834 中的 Ardicrenin 含量为 8.2%，分别比由 A4 诱导的 ACHR 4（4.5%）、组织培养形成的 ACR C（3.1%）、Euphylla（0.8%）和自然形成的 NR C（3.2%）高 1.8、2.7、9.4 和 2.6 倍。总之，获得的 A. crenata 发根系能够比天然植物自然表达更多的 Ardicrenin。
