关键词: 金线莲, 类原球茎, 生物反应器, 金线莲苷

Abstract: To screen the optimal hormone combination and germplasm for inducing Anoectochilus roxburghii protocorm⁃like bodies（PLBs），and to investigate the effects of inoculum density and culture duration on PLBs biomass and kinsenoside content for optimizing large⁃scale culture conditions，this study utilized tissue⁃cultured seedlings of Anoectochilus roxburghii.The effects of different concentration combinations of 6⁃benzylaminopurine（6⁃BA），1⁃naphthaleneacetic acid（NAA），zeatin（ZT），and 2，4⁃dichlorophenoxyacetic acid（2，4⁃D） on the induction of PLBs from five different Anoectochilus roxburghii germplasms were tested.The PLB induction rate，kinsenoside content，and proliferation characteristics were compared among the different plant germplasms.Using a plant bioreactor，the influence of inoculum density（20—80 g/L） and culture duration（10—40 days） on biomass and kinsenoside synthesis was studied.The results indicated that the combination of MS+3 mg/L 6⁃BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L 2，4⁃D medium had the best effect on improving the induction rate of adventitious buds of different species of Anoectochilus roxburghii germplasms.Under identical culture conditions，the ZY1（Nanjing，Fujian） germplasm demonstrated superior PLB induction rate（93%），kinsenoside content（5.03%），and radial proliferation capacity compared to other germplasms.In the bioreactor culture system，both the proliferation coefficient（1.46）and kinsenoside content（7.80%）of the PLBs peaked at an inoculum density of 60 g/L after 40 days of culture in darkness.In conclusion，utilizing the superior ZY1 germplasm with the MS+3 mg/L 6⁃BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L2，4⁃D medium，combined with bioreactor culture conditions of 60 g/L inoculum density and 40 days of culture in darkness，enables the highly efficient accumulation of both Anoectochilus roxburghii PLB biomass and kinsenoside content.

Key words: Anoectochilus roxburghii, Protocorm?like body, Bioreactor, Kinsenoside