李灿 王永清

摘要[目的]研究一种或多种适合春剑（Cymbidum longibracteatum）愈伤组织诱导的方法，并试图找到有效控制褐化的措施。[方法]以春剑花梗为外植体进行离体培养，通过正交设计研究不同基本培养基及激素配比对花梗培养时褐化、生长和愈伤组织发生的影響；同时研究不同防褐化处理对花梗切段褐化率的影响。[结果]不同基本培养基及激素配比对花梗培养的影响显著，不同处理方式对花梗褐化控制效果显著。花梗切段愈伤组织诱导的最佳配方为B5+2，4D 2.0 mg/L+TDZ 0.6 mg/L +GA30.5 mg/L，诱导率为7.32%。当花芽长度为2.0 cm时，花梗褐化率较低；当花梗切割厚度为8.0 mm时，花梗褐化率较低；PVP、VC、CA、8HQS对花梗切段褐化均有显著的抑制作用，其中以8HQS效果最好。[结论]建立了春剑花梗离体培养技术体系，为春剑快速繁殖和遗传育种奠定基础。

关键词春剑；花梗；离体培养；愈伤组织诱导；褐化

中图分类号S604+.3文献标识码A文章编号0517-6611（2016）04-178-05

Study on in vitro Culture of Peduncle in Cymbidium longibracteatum

LI Can， WANG Yongqing* （College of Landscape Architecture， Sichuan Agricultural University， Chengdu， Sichuan 611130）

Abstract[Objective] The aim was to find the best way of callus induction for Cymbidium longibracteatum and the effective measures for browning control. [Method] C. longibracteatum peduncle was used as explants for in vitro culture. The effects of different basic medium and hormones on browning， growth and callus induction were studied through orthogonal design， as well as effects of different browning control treatments on browning in peduncle cutting culture.[Result] Different basic culture mediums and hormone combinations had significant effects on peduncle culture， different treatments had significant controlling effects on browning. The best medium for callus induction from peduncle was B5 +2，4D 2.0 mg/L +TDZ 0.6 mg/L +GA3 0.5 mg/L， resulting in 7.32% of callus induction.When the length of flower bud was 2 cm， the peduncle had low browning rate； peduncle explants of 8 mm thickness had the lowest rate of browning； PVP， VC， Citric acid and 8HQS had significant effects on browning control， but 8HQS displayed best results. [Conclusion] The in vitro culture technology system of Cymbidium longibracteatum peduncle is established， which will lay a foundation for rapid propagation and genetic breeding.

Key wordsCymbidium longibracteatum； Peduncle； in vitro culture； Callus induction； Browning

兰花（Cymbidium ssp.）属于兰科兰花属植物，是我国的传统名花，包括春兰、建兰、蕙兰、墨兰、寒兰等，其中春剑（C.longibracteatum）名品较多，为春兰的变种，稀少而珍贵，而常规的分株繁殖很难满足爱好者的需求。兰花新繁殖技术的探索始于20世纪初，20世纪70年代后，国兰的组织培养在我国兴起。目前已陆续建立了建兰、春兰、蕙兰、墨兰、寒兰等组织培养快速繁殖无性系[1-9]。然而国兰离体培养中褐化问题较严重，对外植体的诱导和继代培养造成巨大障碍。控制褐化的常用措施有取材时间、外植体成熟度[11]、母株或外植体预处理[12]、低温处理[13]、切口封闭[5]、吸附剂和抗氧化剂[14]、热激处理[15]、暗培养[16] ，均可在一定程度上减轻褐化影响。……