国家自然科学基金青年基金（31301250）, 湖北省教育厅科学技术研究项目(B20094008, B2015394)和武汉市高校科学研究资助项目(2008K079)共同资助。
以大花蕙兰原球茎为材料，从培养时间、基本培养基、糖浓度、培养方式及切割方式5个方面探讨了原球茎增殖、分化及离体保存的问题。结果表明，2个月内随着培养时间的延长，原球茎增殖与分化越显著，但培养2个月后，原球茎增殖不明显，分化却持续增加；1/2MS基本培养基利于原球茎增殖与分化，而3MS基本培养基利于原球茎保存；糖浓度的高低对原球茎增殖与分化影响显著，20 g·L-1蔗糖适于原球茎保存，50 g·L-1蔗糖适于原球茎增殖，而原球茎分化成苗应选用30 g·L-1白糖；在3种培养方式中，固体培养利于原球茎分化，液体振荡培养利于原球茎增殖，液体静置培养利于原球茎离体保存；片状切割法利于原球茎增殖与分化，整体剥离接种法利于原球茎保存。即以2.0 mg·L-1 6-BA +0.1 mg·L-1 NAA为激素组合时，以片状切割法接种原球茎于1/2MS+50 g·L-1蔗糖的培养基中并以液体振荡方式培养时利于原球茎增殖；以片状切割法接种原球茎于1/2MS+30 g·L-1白糖的培养基添中并以固体方式培养时利于原球茎分化成试管苗；以整体剥离法接种原球茎于3MS+20 g·L-1蔗糖培养基中并以液体静置方式培养时利于原球茎保存。上述结论为大花蕙兰试管苗快速繁殖与离体保存提供了有效的技术支持。
Using the protocorm-like body (PLB) of Cymbidium hybridum, the effects of culture time, basic medium,?sugar concentration,?cultivation form,?and?cutting?way on proliferation,?differentiation and?conservation in vitro?were studied. The results showed that the PLB?proliferated?and differentiated in two months culture with the differentiation?being more effective after two months culture. Half-strength MS medium?was beneficial to?PLB?proliferation and differentiation,?while 3 MS?medium was favorable to?PLB preservation. Sucrose at 20 g·L-1 was good for?PLB?preservation,?while 50 g·L-1 sucrose?was suitable for?PLB proliferation. Sucrose at 30 g·L-1 was effective for PLB differentiation. Solid culture had an obvious effect for PLB differentiation,?while liquid culture?was good for PLB proliferation.?PLB conservation in vitro could use static liquid culture. The sheet-cutting method?was suitable for?PLB?proliferation and differentiation?and the whole-peeling method was good for?PLB?preservation. The best PLB proliferation condition was 1/2 MS+50 g·L-1 sucrose +2.0 mg·L-1 6-BA +0.1 mg·L-1 NAA in liquid culture with the sheet-cutting method. The best PLB differentiation condition was 1/2 MS+ 30 g·L-1 sugar in solid culture with the sheet-cutting method. The best PLB conservation condition was 3 MS+ 20 g·L-1 sucrose in liquid culture with the whole?peeling method. It indicated that this method is effective for rapid propagation and conservation of Cymbidium hybridum in vitro.