瀕危藥材金線蓮愈傷組織誘導及分化成苗研究

許晴 穆梁麗 冀愛青

摘要[目的]建立有效的瀕危藥材金線蓮的再生體系，為其工廠化生產奠定技術基礎。[方法]選取福建金線蓮的葉片、莖片、帶節莖段、不定芽為外植體，建立無菌體系，而后以愈傷組織的誘導率為指標，采用正交設計優化不同外植體各階段（愈傷組織的誘導和分化）培養基中激素的種類及濃度。[結果]在試驗的福建金線蓮4種外植體中，不定芽的愈傷組織誘導率最高，可達96.67%，其最適培養基配方為MS+0.5 mg/L NAA+4.0 mg/L BA+0.3 mg/L KT。在此培養基上，誘導的愈傷組織在分化培養基MS+0.5 mg/L NAA+2.0 mg/L BA和增殖培養基MS+0.2 mg/L NAA+2.0 mg/L BA上繼代形成了無根試管苗。[結論]該研究建立的無菌繁殖體系，解決了前人所指出的金線蓮愈傷組織誘導率極低的瓶頸問題，組培成苗的效率有較大提高。

關鍵詞金線蓮；不定芽；愈傷組織；分化成苗

中圖分類號S567.23+9文獻標識碼A文章編號0517-6611（2017）14-0103-04

Abstract[Objective] The research aimed to establish an effective recycling system for Anoectochilus roxburghii， and lay the technical foundation for its industrial production. [Method] As explant of leaf， stem， stem segment with node and adventitious bud of Anoectochilus roxburghii， the aseptic system was established， and then the induction rate of callus was used as an index to optimize the type and concentration of hormones in the culture medium of different explants （callus induction and differentiation） by orthogonal design.[Result]In the test of Anoectochilus roxburghii， four explants， the callus induction rate of adventitious bud was the highest， up to 96.67%， the optimum medium formula was MS+0.5 mg/L NAA+4.0 mg/L BA+0.3 mg/L KT. On the basis of the medium， the callus was subcultured on the differentiation medium（MS+0.5 mg/L NAA+2.0 mg/L BA） and the proliferation medium （MS+0.2 mg/L NAA+2.0 mg/L BA） to form a rootless test tube.[Conclusion]Asepsis reproductive system has been established， which solves the bottleneck problem that the predecessors pointed out that the callus induction rate of Anoectochilus roxburghii was very low，the efficiency of tissue culture seedlings have greatly improved.

Key wordsAnoectochilus roxburghii；Adventitious buds；Callus；Seeding differentiation

金線蓮[Anoectochilus roxburghii（Wall.） Lindl]，別名金絲線、金耳環等，蘭科開唇蘭屬多年生草本植物。金線蓮是我國珍稀瀕危的名貴藥材，素有“藥中之王”之稱，其全草均可入藥，有清熱涼血、祛風利濕、解毒、止痛、鎮咳等功效。近年來，金線蓮在治療高血壓、糖尿病及腫瘤等疑難病癥方面日益引起醫藥界的重視，具有廣闊的開發利用前景。金線蓮的種子不易萌發，用傳統的實生繁殖相當困難，且其栽培對生態環境要求嚴格，加之自然環境遭受嚴重破壞，生長極其緩慢，且蟲鳥等動物喜食，自然資源稀少，導致市場價格昂貴，高達3 000～5 000元/kg，不利于金線蓮的推廣。而植物組織培養技術作為一種無性繁殖的手段，可在短期內提供大量優質的試管苗，使其大面積推廣成為可能?！?br>