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

摘要： 筛选金线莲类原球茎（Protocorm⁃like bodies，PLBs）诱导的最佳激素配比和种源，探究接种量与培养时间对PLBs生物量及金线莲苷含量的影响，以优化金线莲PLBs规模化培养条件。以金线莲组培苗为试材，测试不同质量浓度6-苄基氨基嘌呤（6-BA）、萘乙酸（NAA）、玉米素（ZT）和2，4-二氯苯氧乙酸（2，4-D）组合对5种金线莲PLBs诱导的影响，比较不同种源PLBs诱导率、金线莲苷含量及增殖特征；利用植物生物反应器，研究接种量（20~80 g/L）和培养时间（10~40 d）对生物量及金线莲苷合成的影响。结果表明，MS+3 mg/L 6-BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L 2，4-D培养基组合提高不同种金线莲不定芽诱导率效果最佳。同等培养条件下，ZY1（福建南靖）种源的PLBs诱导率（93%）、金线莲苷含量（5.03%）及径向增殖能力均优于其他种源。在生物反应器培养系统中，接种量60 g/L、暗光培养40 d时，金线莲PLBs增殖系数（1.46）和金线莲苷含量（7.80%）均达到峰值。综上，利用ZY1优势种源，基于MS+3 mg/L 6-BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L 2，4-D培养基，结合接种量60 g/L、暗光培养40 d的生物反应器培养条件，可实现金线莲类原球茎生物量与金线莲苷含量的高效积累。

关键词: 金线莲, 类原球茎, 生物反应器, 金线莲苷